KUMPULAN ABSTRAK | CONTOH JUDUL PENELITIAN MIKROBIOLOGI

Teknologi Pupuk Mikrob Multiguna Menunjang Keberlanjutan Sistem Produksi Kedelai

(Technology of Multipurpose Microbial Fertilizer Supporting Sustainable System of Soybean Production)

RASTI SARASWATI

Balai Penelitian Bioteknologi Tanaman Pangan, Jln. Tentara Pelajar 3A, Bogor 16111; Tel. 062-251-337975 pes. 224; Faks. 062-251-338820

Multipurpose microbial fertilizer (MPMF) has been developed to increase fertilization efficiency to support sustainable soybean production system. Multipurpose microbial fertilizer can supply a substantial portion of nitrogen and phosphorous which is required by soybean through their symbiotic relationship rhizobia and phosphate dissolving capability, and thus save the us of anorganic fertilizer. Result of the demonstration plot using MPMF in farmer fields showed accordingly that phosphate fertilizer up to 50% from the recommendation dosage.

Key word: microbial fertilizer, inoculant, soybean

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Pemberian Inokulan Campuran Beberapa Cendawan Mikoriza Arbuskula pada Kacang Tanah dan Kedelai

(The Application of Mixed Arbuscular Mycorrhizal Fungi Inoculant to Peanut and Soybean)

M. RAHMANSYAH & SUCIATMIH

Puslitbang Biologi LIPI, Jln. Ir. Juanda, Bogor 16122 ; Tel. 062-251-324006, Faks. 062-251-325854

Glomus sp.4, as arbuscular mycorrhizal (AM) fungi, was a collection originally gathered from soil in Bogor and had been introduced succesfully to fast-growing legumes of Albizia procera, Paraserianthes falcataria and pterocarpus indicus seedlings. To determine the influence of AM fungi inoculation to the plant growth, the experiment was set up as follow, a. Glomus sp.4 was mixed with G. etunicatum, G. manihotis and G. microagregatum+acaulospora spinosa, and inoculated separately as A, B, and C soil culture inoculant.; b. peanut and soybean were inoculated and planted as pot experiment in mixed medium of soil, compost and sand (2:1:1); c. for controlling treatments the plants were not inoculated and the other one were planted at medium which was amanded with 100g/kg TSP. As plant growth, AM fungi colonies of both plants roots increased. Fungi infection were founded higher in soybean (50-90%) compared to peanut infection (40-70%). Peanut shoot dryweight (15.6 g) and index of seed over shell (1.01) whiwh were harvested 53 days after planting (DAP) showed value as caused by B inoculant. Soybean plants at 45 DAP were significantly different in shoot dry weight (2.08 g) and seed weight plant-1 (4.12 g) compared with control, as caused of C inoculant treatment. Phosphorus content in dry shoot of peanut treated with B inoculant was 0.3%, and soybean which C inoculant was 0.4%.

Key word: inoculant, arbuscular mycorrhizal fungi, peanut, soybean

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Proses Fermentasi Biji Lamtoro-Gung Dengan Rhyzopus oryzae

(Fermentation Process of Leucaena Seed with Rhyzopus oryzae)

KOMARI

Pusat Penelitian dan Pengembangan Gizi, Jln. Dr. Semeru, Bogor 16122

Fermentation process of leucaena seeds with Rhyzopus oryzae was developed to study biochemical during fermentation process with the emphasis on iron evailability (in vitro). Fermentation significantly increased the solubilities of protein and carbohydrate (soluble sugars). The increas of tannins content of the seeds during fermentation was due to loss of binding capacity between tannins and protein or carbohydrate. The loss was due to the increase in the pH value and the decrease in size of the digested protein and carbohydrate by the activity of enzymes produced by microorganism. Although the detectable tannins in the leucana tempe was higher than that in unfermented seeds, there was no binding effect of tannins with iron. Iron availability of the leucana tempe increased from 1.9% in the cotyledons to 11.6%. This finding showed the benefit effect of fermentation on the iron availability of the tempe.

Key word: fermentation process, leucana seed, Rhyzopus oryzae

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Penggunaan “MicurR-BT” sebagai Uji Awal Sebelum Pembiakan Spesimen Urin untuk Isolasi Etiologi Infeksi Saluran Kemih

(MicurR-BT As an Indicator of Antibiotic in the Urine Before Bacterial Isolation in Urinary Tract Infection)

PRATIWI SUDARMONO & TERTIA HUTABARAT

Bagian Mikrobiologi FK UI, Jln. Pegangsaan Timur No. 16, Jakarta 10320, Tel. 062-021-310086, Faks. 062-021-3100810

Urinary tract infection is very common and has a very high indence rate in Indonesia. Usually it cause by bacterial infection, so the diagnostic microbialogy play a very important role in the management of antibiotic therapy. Unfortunately, the patients usually has taken antibiotic before examination. MicurR-BT is a dipstick test to detect the existence of antibiotic in urine. It contains Bacillus subtilis and triphenyl tetra zolium chlorade as color detector. Ninety six urine specimen has been tested before bacterial culture. Fifty four specimen (56%) contain antibiotic in the urine before culture.

Key word: urinary tract infection

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Deteksi Virus Dengue Tipe 2 dengan Cara Hibridisasi in situ

(Detection of Tipe 2 Dengue Virus by in situ Hibridization)

MAKSUM RADJI, AMIN SOEBANDRIO, MIRAWATI SUDIRO & PRATIWI SUDARMONO

Bagian Mikrobiologi FK UI, Jln. Pegangsaan Timur No. 16, Jakarta 10320 Tel. 062-021-3100806, Faks. 062-021-3100810, Email: amin0207@rad.net.id

Demonstration of dengue virus (DV) in infected cell wouold enable elaboration of pathogenesis and pathophysiology of dengue Fever and dengue Haemorrhagic Fever. Molecular detection of DV would give high sensitivity as well as specifity. C6/36 mosquito cell line artificially infected with DV was used as a model of DV infected cell. 290-bp cDNA of envelope region of DV type 2 (DV-2) labeled with digoxigenin-11-dUTP was used as probe. Hybridization was performed directly to infected cell fixed on to glass slide (in situ). 10 ng/ul of the probe was able to detect as low as 10x TCID 50 infecting DV-2. The signal produced was not found in negative control and was clearly increasing in infecting viral dose dependent manner. There was no cross reactivity between DV-2 probe and DV-3 and vice versa. The DV-2 probe was sensitive yet specific in demonstrating the presence of DV-2 in infected cell.

Key word: hybridization, digoxygenin (DIG), pathogenesis

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Campuran Kapas dan Kelaras Pisang sebagai Media Tanam Jamur Merang

(Mixture of Cotton Waste and Dried Banana Leaves as the Media for Straw Mushroom Cultivation)

METTY IRAWATI, AGUSTIN WYDIA GUNAWAN & OKKY SETYAWATI DHARMAPUTRA

Laboratorium Mikologi, Jurusan BiologiFMIPA IPB, Jln. Raya Pajajaran, Bogor 16144

In Indonesia straw compost is used as common medium for straw mushroom cultivation, because its high cellulose and hemicellulose content. Never theless, waste cotton derived from textile industry and dried banana leaves can be used for straw mushroom cultivation, because their cellulose and hemicellulose content is also high. Cotton waste and dried banana leaves were composted for 20 days by adding 2% of lime and 8% of rice bran. The compost of cotton waste, dried banana leaves, and mixture of cotton waste and dried banana leaves ratio of 4:1, 3:1, and 1:1 were used as the media for straw mushroom cultivation. Three replications were used for each treatment. The media were pasteurized at about 60oC for two hours, further the temperature was maintained at 50oC for 10 hours. Spawning was carried out when the temperature dropped to 30oC, and then the mushroom house was closed for three days. This condition was necessary for mycelial growth. After that, fresh air was introduced into the house of basidioma formation and development. Harvesting was carried out when the basidioma was at button or egg stage. Mushroom production on the mixture of cotton waste and dried banana leaves at a ratio of 1:1 was not significantly different than cultured on cotton waste only. The production on the other mixture were higher and significantly different than those cultured on cotton waste or banana dried leaves.

Key word: straw mushroom, media for straw mushroom cultivation, cotton waste, dried banana leaves

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Penggunaan Kromatografi Tukar Ion DEAE-Spharos CL-6B Dalam Purifikasi Komponen Selulase Cellulomonas CS1-17

(The Use of Ion Exchange Chromytography of DEAE-Sepharose CL-6B in the Purification of Cellulase Components from Cellulomonas CS1-17)

M.B. TRESNAWATI PURWADARIA

Balai Penelitian Ternak, Kotak Pos 221, Bogor 16002 Tel. 0251.240752, Faks. 0251.240754, Email: Balitnak@indo.net.id

Ion exchange chromatography of DEAE-sepharose CL-6B was used to purify cellulase components from a culture filtrate of Cellullomonas CS1-17 grown on microcrystalline cellulose (Sigmacell-20). The cellulase components were separated into three activity peaks after elution with a linear gradient of NaCl concentrations from 0 to 0.5M. The first and second peaks (Ia and Ib) contained high activity for degradation of amorphous celllulose (CMCase), while the third peak (Ic) contained activity for degradation of microcrystalline cellulose (avicelase). In the higher concentration of NaCl (0.0 to 1.0M) and in the same linear gradient addition the filtrate was separated into four peaks. Three peaks of IIa, IIb, and IIc had the same activity as Ia, Ib, and Ic, while the last peak (IId) had high CMCase activity. In this separation the resolution between IIb and IIc was poorer than Ia and Ib. Therefore, in the further experiment the separation was carried out with stepwise gradients of NaCl (0.008, 0.18, 0.30 and 1.0 M). The choice of salt concentrations was based on results of the second linear gradients system. Five peaks were resolved well after the elution. Further enzyme determination shows that IIb has high CMCase activity, IIc has high avicelase activity, while IId has cellobiohydrolase activity.

Key word: ion exchange chromatography, purification of cellulose components, Cellulomonas CS1-17

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Laboratory Prescreening of Bradyrhizobium japonicum for Acid pH Tolerance to Predict Their Survival in Acid Soils

ARIEF INDRA SUMUNAR1 & P. J. DART2

1Research Institute for Food Crops Biotechnology, Jalan Tentara Pelajar No. 3A, Bogor 16111
2School of Land and Food, The University of Queensland, St. Lucia, Australia

Introduced Bradyrhizobium japonicum strains are often unable to tolerate acid soil stress factors in a new environment. There is a need to improve the survival of B. japonicum in acid soils. This experiment was conducted to study the selectivity of four agar media in screening acid tolerant strains. Sixteen strains of B. japonicum were stab inoculated onto four screening media at five pH levels (3.8, 4.2, 4.5, 5.0, and 6.8) and the growth of each starin was observed and scored everyday. To test the usefulness of this screening method, B. japonicum strains were inoculated to two sterile acid soils which differ in chemical properties. The survival of each strain was determined at days 1, 8, 18, and 28 after inoculation. The results showed that the selectivity of each screening medium was different, where the more acid stress factors incorporated to that medium the more selective was that medium. The relationship between screening in acidic agar media and survival capability in sterile acid soils was quite well established, where the strain that survived well or poorly in sterile soils were those that were identified as acid tolerant or acid sensitive strains in acidic agar media. Results from this experiment confirm that laboratory prescreening of B. japonicum for acid, Al and Mn tolerance using acidic agar media was succesful in selecting strains which were tolerant in low pH soils, and those which were less so.

Key words: laboratory prescreening, Bradyrhizobium japonicum, screening media, acid soil

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Amonifikasi Kulit Buah Kakao sebagai Tindakan Alternatif untuk Memusnahkan Inokulum Phytophthora palmivora

(Ammonification of Cocoa Husks as an Alternative Method to Eliminate Phytophthora palmivora Inoculum)

T. W. DARMONO1, TRI PANJI1 & H. KWARTONO2

1Unit PenelitianBioteknologi Perkebunan, Jalan Taman Kencana No. 1, Bogor 16151
2Universitas Pakuan, Bogor

Cocoa husks is produced in a large quantity and may cause a great deal of problem if not managed properly. Cocoa husks buried in soil or laid on the ground are easily infested by Phytophthora palmivora and become a potential source of pod rot disease inoculum. Pod rot is one of the most important disease in cocoa. The disease is difficult to control because its source of inoculum is difficult to be eliminated. Gaseous ammonia produced from ammonification is known to be toxic to several microbes and may be used to suppress the inoculum potential of P. palmivora in cocoa husks. The goal of this study was to determine the potential use of ammonification process in the elimination of P. palmivora inoculum. Ammonification of cocoa husks was conducted by mixing cocoa husks with urea in a closed container. Cocoa husks was inokulated with P. palmivora before there were treated with urea. Levels of the inoculum potential were dtermined using bioassay technique. The level of N total in cocoa husks and the level of N ammonia were determined using Kjeldal technique. From this study it was found that a significant amount of ammonia was produced during ammonification process of cocoa husks with the addition of urea. The ammonia produced eliminated P. palmivora inoculum in cocoa husks tissue within one week at an application level of 0.4% urea. Application of ammonification technique for the eradication of P. palmivora was proven to be effective in the field at 2% urea. Besides free from P.palmivora, the cocoa husks became 29.5% richer of nitrogen after ammonification with 0.2% urea.

Key words: ammonification, Phytophthora, cocoa

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Konstruksi Mutan Protein Disulfida Isomerase pada Saccharomyces cerevisiae

(Construction Protein Disulphide Isomerase Mutant on Saccharomyces cerevisiae)

JUMIARTI AGUS

Balai Penelitian Bioteknologi Tanaman Pangan, Jalan Tentara Pelajar No. 3A, Bogor 16111 Tel. 62-251-337975 pes. 224 Faks. 62-251-338820

Protein disulphide isomerase (PDI) is an enzyme that catalyses disulphide-bond formation in protein to maintain the native conformation, either through redox or isomerization reactions. Few PDI’s have been isolated from various organism such as mammalian livers, plants, green algae and Saccharomyces cerevisiae. Disruption of the PDI in S. cerevisiae is haplo lethal indicating that the product of this gene is essential for viability. PDI consist of 1590 pb, but the essential domain on PDI gene have not been known complete. To study caracteristic PDI protein domains, in this research was undertaken construstion PDI mutant on S. cerevisiae. In vitro mutagenesis was carried out using hidroxilamin (HA) as mutagen. Recombinant plasmid were constructed by ligation of mutated DNA fragment (758 pb) into a vector (6300 pb) using T4 DNA ligase. The result showed some mutants were lethal, indicated that b and b’ domains are essential for PDI on S. cerevisiae. Few mutants poorly growth, and the other showed well growth.

Key words: construstion, protein disulfide isomerase, Saccharomyces cerevisiae

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Studi Resistensi Neisseria gonorrhoeae yang Diisolasi dari Pekerja Seks Komersial di Beberapa Tempat di Jakarta

(Antimicrobial Susceptibility Pattern of Neisseria gonorrhoeae isolated from Female Commercial Sex Workers in Jakarta)

YEVA ROSANA1, Agus Sjahrurachman1, Endang R. Sedyaningsih2, Cyrus H. Simanjuntak2, Sumaryati Arjoso2, Sjaiful Fahmi Daili3, Jubianto Judanarso1 & Ika Ningsih1

1Bagian Mikrobiologi, Fakultas Kedokteran, Universitas Indonesia, Jalan Pegangsaan Timur 16, Jakarta 10320
2Pusat Penelitian Penyakit Menular, Badan Penelitian dan Pengembangan Kesehatan DepKes RI
3Bagian Ilmu Penyakit Kulit dan Kelamin, Fakultas Kedokteran, Universitas Indonesia

The control sexually transmitted diseases (STDs) of infection has become more urgent since it has been documented that untreated STDs infection facilitates HIV transmission. One of the most frequent cause of STDs reported is Neisseria gonorrhoeae. Problem of gonorrheal infection is complicated by the occurrence of resistant strains all over the world. In this study, gonococcal isolation have been attempted from 165 endocervical swabs from Female Commercial Sex Workers (FCSWs) in Kramat Tunggak and Kedoya Jakarta. Of 114 Kramat Tunggak’s samples examined, 61 (53.5%) were positive for gonococcal cultures, whereas 13 (25.5%) from 51 Kedoya’s sample were positive. The penicillinase-producing N. gonorrhoeae of gonococcal isolates from Kramat Tunggak and Kedoya were 73.8% and 92.3%, respectively. The result of the Minimun Inhibitory Concentration tests for 6 antimicrobials (thiamphenicol, kanamycin, spectinomycin, ciprofloxacin, cefuroxime, and ceftriaxone) shows that 1.6% isolates were resistant and 34.4% isolates were intermediate to the kanamycin, and 1.6% isolates were intermediate to the thiamphenicol, and all isolates were sensitive to the spectinomycin, ciprofloxacin, cefuroxime, and ceftriaxone. Antibiotic with best activity in vitro was fluoroquinolone (ciprofloxacin). Whereas antimicrobial activity of the cephalo-sporins (ceftriaxone, and cefuroxime) were better than spectinomycin, thiamphenicol, and kanamycin.

Key word: Neisseria gonorrhoeae, female commercial sex workers, minimum inhibitory concentration

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Proses Fermentasi Fed-Batch Untuk Produksi Dekstranase dengan Streptococcus sp. B7

(Fed-Batch Fermentation Processes to Produce Dextranase from of Streptococcus sp. B7)

BUDIATMAN SATIAWIHARDJA1, BENI WIBISONO2 & UNTUNG MURDIYATMO2

1Jurusan Teknologi Pangan dan Gizi, Fateta, Institut Pertanian Bogor, Kampus IPB Darmaga, Bogor 16680
2Pusat Penelitian Perkebunan Gula Indonesia, Jalan Pahlawan No. 25, Pasuruan 67126

The research aimed to develop fermentation technique on dextranase production from the bacterial isolate of Streptococcus sp. B7 using fed-batch fermentation system. The production was done in two litre fermentor with flow rate of medium addition at 19 ml/hr after 24 hours until 72 hours of incubation process. The variable conditions were pH of 7 and 8 and agitation speeds of 300 and 500 rpm. Maximum production was achieved at 500 rpm and pH 8 that produced enzyme activity of 920 U/ml. The best result of some kinetic parameters were as follows : production p = 920 U/ml, productivity Qv = 16.43 U/ml/hr, dextranase specific activity 17.68 U/mg protein, yield of product over cell Yp/x = 696.47 U/g cell and specific productivity qp = 12.45 U/g cell/hr.

Key words: dextranase, fed-batch fermentation, Streptococcus sp. B7, kinetic parameters

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Perilaku Kultivasi Isolat Bakteri Termofil Penghasil a-Amilase

Cultivation of Thermophilic Bacteria Isolate of a-Amylase Production

NUR RICHANA, GAGAN MAULANA YUSUF, PUJI LESTARI & DJOKO SAID DAMARDJATI

Balai Penelitian Bioteknologi Tanaman Pangan, Jalan Tentara Pelajar No. 3A, Bogor 16111

Cultivation of thermophilic bacteria isolate TVII-6 for a-amylase production. Thermophilic bacteria TVII-6 was isolated and selected from the soil sample taken from Dieng vulcanic. The specific activity of amylase produced by TVII-6 was 1624.37 U/mg protein. Cassava starch was used as carbon source on enrichment culture and the optimum concentration was 1% with specific activity 2092.23 U/mg protein. Cultivation in a 2-l bioreactor at pH 7.0 and temperature of 50oC, combine with 250 rpm agitation and 1.0 vvm aeration produced the highest a-amylase. The maximum specific growth rate obtained was 0.147/hours. The relationship between product and substrate was linear (Y=0.129 X + 0.024). Efficiency of the substrate to produce amylase (Yp/s) was 0.129 g protein/g substrate. Relationship between the bacterial growth and substrate was Y = 0.315 X + 0.013. Efficiency of the substrate for growing the bacteria (Yx/s) was 0.315 g cell/g substrate. Based on the bacterial growth and amylase production, amylase could be considered growth associated, with kinetic parameter of product rate (p) was 0.37g/g/hr.

Key words: thermophilic bacteria, a-amylase

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The Effects of Xanthorrhizol on the Morphology of Candida Cells Examined by Scanning Electron Microscopy

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Department of Biotechnology, Yonsei University, 134-Sinchon-dong, Seodaemun-gu, Seoul 120-749, Korea

The effects of xanthorrhizol, a natural anticandidal agent isolated from the rhizome of temulawak or java turmeric (Curcuma xanthorrhiza Roxb.) on the morphology of four human pathogenic Candida species, i.e., C. albicans, C. glabrata, C. guilliermondii, and C. parapsilosis was examined by scanning electron microscopy (SEM). The SEM analysis showed that, unlike control cells representing normal oval to spherical with smooth surface, treatment of Candida strains with xanthorrhizol at 1 x MICs (minimum inhibitory concentration) significantly affected the external morphology, exhibiting deformation, and protrusions on the cell surface. The potent anticandidal activity of xanthorrhizol may support the use of medicinal plants for the treatment of candidal infections.

Key words: anticandidal, Candida sp., scanning electron microscopy, xanthorrhizol

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Kajian Teknik Kultivasi dan Pengaruh Luas Permukaan Media Tumbuh pada Produksi Selulosa Menggunakan Bakteri Isolat Lokal

Study on the Cultivation Technique and the Effect of Surface Area of Growth Media to the Production of Cellulose by Indigenous Isolates

ANI SURYANI1, ANDES ISMAYANA1, YENITA SUATRINA1 & YU-RYANG PYUN2

1Pusat Antar Universitas Bioteknologi, Institut Pertanian Bogor, Kampus IPB Darmaga, Kotak Pos 1, Bogor 16610
2Bioproduct Research Center, College of Engineering, Yonsei University, Seoul 120-749, Korea

RBP-52 and PD5 isolates were isolated from Indonesian fruits samples that contains glucose. RBP-52 isolate was obtained from coconut water (waste), collected from nata factory, and PD5 isolate was obtained from rotten coconut sample, collected from Parung. They were identified as Acetobacter liquefaciens. RBP-52 and PD5 isolate produced cellulose in static and shaking cultivation. Production of cellulose by RBP 52 and PD5 were optimized in static cultivation. Cellulose was found to be produced at the liquid surface in static cultivation. The rate of cellulose production depended proportionally on the surface area of the culture medium. The maximum production rate of cellulose was 2.78 g/l per 7 days (surface area 77.60 cm2) in the static cultivation.

Key words : Bacterial cellulose, static culture, shaken culture, RBP-52 and PD5 isolates, surface area

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Pemurnian ?-Amilase Bacillus stearothermophilus dengan Membran Ultrafiltrasi

Purification of ?-Amylase from Bacillus stearothermophilus by Ultrafiltration Membrane

PUJI LESTARI1, NUR RICHANA1 & UNTUNG MURDIYATMO2

1Balai Penelitian Bioteknologi Tanaman Pangan, Jalan Tentara Pelajar No. 3A, Bogor 16111
2PTPN XI Surabaya, Jalan Merak No. 1, Surabaya

The objective of this experiment was to evaluate the optimum conditions of a-amylase of Bacillus stearothermophilus purification by ultrafiltration and the step of a-amylase purification from indigenous isolate. Culture filtrate from bioreactor was separated by microfiltration membrane with a pore size of 0.2 mm. The supernatant was purified again by ultrafiltration system (membrane with cut off 30 000 Dalton). Treatments of a-amylase purified by ultrafiltration were carried out at flow rate of 30, 45 and 60 ml/minute, and concentrated by about 5, 10 and 15 times. The crude enzymes resulted from ultrafiltration were precipitated with acetone. The results showed that the optimum condition of ultrafiltration was using flow rate of 30 ml/minute and concentrated by about 10 times. At the optimum condition of ultrafiltration, the specific activity of a-amylase was of 6 686.6 U/mg with 2.3 fold purification factor. The effect of flow rate decreased the total enzyme activity, specific activity and yield. The concentration disposal could decrease total activity and protein, but not always reduced specific activity of the enzyme. Purification of crude enzyme by ultrafiltration and acetone reduced the total activity, total protein and yield, but specific activity and purification factor increased. Ultrafiltration followed by acetone precipitation, gave enzyme specific activity of 18155.4 U/mg, purification factor of 6.3 and yield of 20%, respectively. Zymogram analysis using Native-Polyacrylamide Agarose Gel Electrophoresis indicated a-amylase of approximately 192 932.8 Da.

Key words: a-amylase, Bacillus stearothermophilus, ultrafiltration

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Biodegradasi Senyawa Epiklorohidrin oleh Bakteri Isolat G3

Biodegradation of Epichlorohydrin Compound by Bacterial Isolate G3

ANAS MIFTAH FAUZI, AMIN PRIYO UTOMO, NITARIANI ELFRIDA & MAYA NILASARI

Center for Development of Safe Agroindustrial Processes, Gedung Fateta Institut Pertanian Bogor, Kampus IPB Darmaga, Bogor 16002

Biodegradation of epichlorohydrin (ECH) by strain G3 (temporarily identified as Pseudomonas aeruginosa) was investigated both in suspended cultures and biofilm reactors. Experiment was performed to study biodegradation profile of the target pollutant in the presence of other organic pollutants, i.e. mixture of benzene, toluene, xylene, and heavy metal, i.e. Pb(NO3)2. The results indicated that the chemicals reduced the specific growth rates of G3 isolate in the suspended cultures with ECH added as the sole carbon source. These chemicals also slightly reduced dechlorination rate of ECH in the effluent of biofilm reactor. This study also showed that the magnitude of 2 mg/l ECH degradation decreased with the lower retention time. 67.6% of degradation was occured during 1.33 min retention time.

Key words: epichlorohydrin, biofilm reactor, biodegradation

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Pengaruh Pemberian Rhizo-Plus pada Kedelai

Influence of Rhizo-Plus Application on Soybean

Ig.V. SUTARTO1 & RASTI SARASWATI2

1Pusat Penelitian dan Pengembangan Tanaman Pangan,
2Balai Penelitian Bioteknologi Tananaman Pangan, Jalan Tentara Pelajar No. 3A, Bogor 16111

The soybean [Glycine max (L.) Merrill] has been recognized as an important source of protein for both food and feed in Indonesia. Among the food legumes, soybean is the first priority in farm operation. Notwithstanding, the mayor constraints for low soybean production are due to drought, pests, and used of traditional cultivation methods. The objective of this experiment was to examine the effect of Rhizo-plus inoculation on the root nodule performance, plant growth. yield of soybean and the economical profit. Field experiment was conducted in farmer’s fields in Temanggung (Central Java) which is typified by Regosol soils during planting season of 1996/1997. The design of these experiment was a split-plot design with four replications. The results showed that the Rhizo-plus inoculation did not interact with the parameters, and it was required to increase the yield of soybean. Rhizo-plus which is produced nationally demonstrated beneficial effect on some plant growth parameters such as N accumulation, the number and weight of root nodules, leaf chlorophyll content, and dry yield of soybean. Soybean cultivar ‘Slamet’ was found to demonstate the best yield. Therefore, this technology might be useful to increase the income and net profit of soybean farmers.

Key words: Rhizo-plus, soybean, Regosol Soils, Central Java

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Berbagai Cara Hidrolisis Pati untuk Media Pertumbuhan Bacillus sp. BMN14 Penghasil Biosurfaktan Lipopeptida

Hydrolysis Methods of Starch for Culture Media of Bacillus sp. BMN14 Producing Biosurfactant Lipopeptide

NUR RICHANA1, ANI SURYANI2, HELENA YUSUF MAKAGIANSAR2 & TUN TEDJA IRAWADI2

1Balai Penelitian Bioteknologi Tanaman Pangan, Jalan Tentara Pelajar No. 3A, Bogor 16111
2Jurusan Teknologi Industri, Fateta IPB, Bogor 16002

Ezymatic and acid hydrolysis of starch for culture media of Bacillus sp. BMN14 producing lipopeptide biosurfactant were studied using cassava, arrowroot and sago starch. These starch hydrolysates were used as glukose substitute in the were to culture media of biosurfactant-producing bacteria. Starch hydrolyzed from arrowroot (73.08-83.5%) and cassava (80.0-87.98%) yielded glucose higher than sago (62.5-64.5%). Capability of the bacteria to produce biosurfactant represented as the amount of acid precipitate (0.78 g/l) and surface tension of the broth (33.6 mN/m) in the culture medium contained enzymatic hydrolyzed-starch.

Key words: lipopeptide biosurfactant, starch hidrolyzed, Bacillus sp.

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Kerentanan Poliester Alifatik Terhadap Biodegradasi

Biodegradability of Aliphatic Polyester

EVITA CHRISNAYANTI1, EFRIDA MARTIUS1, ROFIQ SUNARYANTO1, LIES DWIARTI1, HARDANING PRANAMUDA1 & YUTAKA TOKIWA2 1Pusat Pengkajian dan Penerapan Bioteknologi Industri dan Pertanian, Kawasan Puspiptek, Serpong, Tangerang 15314
2National Institute of Bioscience and Human-Technology, Tsukuba, Ibaraki 305-8566, Japan

The presence of microbes which can degrade commercial biodegradable plastics, i.e. poly-?-caprolactone (PCL), poly-?-hydroxy butyrate (PHB), polybutylene succinate (PBS) and polylactic acid (PLA), was evaluated using colony-counting and clear-zones methods. Out of 12 soil samples taken from Serpong area, it was confirmed that PCL, PHB and PBS degrading microorganisms were observed in all samples, but no samples showed PLA degradation. The ratio of degrading microorganisms to total microorganisms decreased following the order of PCL, PHB, PBS and PLA. Result of burial test of biodegradable plastic films show that PHB film is easy to degrade but not PLA film. The tendency of the results was similar with the results reported in the same investigation at sub-tropical country.

Key words: plastic, polyester, biodegradation

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Analisis Keragaman Kapang Pencemar Pakan Unggas Komersial

The Diversity Analysis of Moulds Contaminating Commercial Poultry Feed

SRI HANDAYANI & JOKO SULISTYO

Balitbang Mikrobiologi, Puslitbang Biologi Lembaga Ilmu Pengetahuan Indonesia, Jalan Ir. H. Juanda No. 18, Bogor 16002

Samples of poultry feed were collected from several poultry shops around Jakarta, Bogor, and Bandung, which represented different location altitudes. Samples were inoculated onto Sabouraud’s glucose agar then incubated at 25 and 37OC to determine kinds and populations of moulds which potentially contaminate the feed. The experimental analysis for different altitudes and incubation temperatures was carried out using factorial completely design 3 x 2 with three replicates. Results indicated that there were 14 different kinds of moulds recovered from all samples. Location altitudes and incubation temperatures did not significantly influenced (p>0.05) the kinds of moulds, however, significantly influenced (p0.05) the kinds of moulds, however, significantly influenced (p90%) percentage of AM fungi colonization. Arum-type of AM colonization found on 13 species of weeds, while Paris-type AM colonization were found on 10 species. Intermediate type of arbuscules were only found on 6 spesies. In this study we also examined the presence of vesicles.

Key words: arbuscular mycorrhizae, arum type, paris type, weed

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Metode PCR Sederhana Untuk Menapis Isolat Bacillus thuringiensis yang Membawa Gen Cry V

A Simple PCR Method for Screening Bacillus thuringiensis Isolates Carrying Cry V Gene

BAHAGIAWATI, DANY SATYAWAN & SUTRISNO

Balai Penelitian Bioteknologi dan Sumberdaya Genetik Pertanian, Jalan Tentara Pelajar No. 3A, Bogor 16114

This experiment was conducted to obtain a simple, rapid and cheap PCR technique that accurately detects the present of cry on isolates of Bacillus thuringiensis (Bt). Further, the technique was applied to identify Indonesia Bt isolates. DNA from direct bacterial colony was used as template for the new PCR technique, instead of plasmid DNA that was used as template for existing methode. Consequently, DNA extraction procedure can now be omitted and the screening process will be tremendously speed up. One hundred sixty-five Bt isolates were screened for the present of cry V and sixteen isolates showed the presence of cry V. Two isolates out of 16 labeled as Jtg 2151 and Jtm 1842 produced DNA bands at approximately 726 base pairs. The band was the expected size of cry V amplicon.

Key words: Bacillus thuringiensis, cry V, PCR techniques

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Keragaman Genetika Bakteri Tanah dari Rizosfer Kapas Transgenik dan Nontransgenik di Soppeng, Sulawesi Selatan

Soil Bacterial Genetic Diversity from Rhizosfer of Transgenic and Nontransgenic Cotton Plantation in Soppeng, South Sulawesi

MUHAMMAD YUSUF1, YUSMINAH HALA2 & ANTONIUS SUWANTO1,3

1Seameo-Biotrop, Jalan Raya Tajur, Bogor 16001
2Jurusan Biologi, FMIPA, Universitas Negeri Makassar, Jalan Daeng Tata Raya, Makassar 90224
3Jurusan Biologi, FMIPA, Institut Pertanian Bogor, Bogor 16144

Techniques based on amplification of 16S-rRNA genes for comparing bacterial communities are now widely used in microbial ecology. In this study, we compared bacterial genetic diversity of transgenic and nontransgenic cotton plantation soil samples to examine the effect of transgenic cotton on soil bacterial diversity. The primer 63f and 1387r specific for bacteria were used to amplify DNA extracted from two soil samples. The PCR products were cloned into pGEM-T Easy and transformed into Escherichia coli DH5-?. Total transformants of transgenic and nontransgenic obtained from cotton plantation soil samples were 138 and 123 respectively. Twenty transformants containing 16S-rRNA genes were selected randomly from each library to reveal their amplified ribosomal DNA restriction analysis (ARDRA) patterns employing restriction enzymes HhaI, RsaI, and HaeIII. The results indicated that there were 16 and 14 different ARDRA profiles derived from transgenic and nontransgenic cotton plantation, respectively.

Key words: bacterial diversity, ARDRA, transgenic and nontransgenic cotton

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Mikoriza Durian di Bogor dan Sekitarnya

Mycorrhiza of Durio in Bogor Area

CHAIRANI1, AGUSTIN WYDIA GUNAWAN1 & KARTINI KRAMADIBRATA2

1Jurusan Biologi, FMIPA, Institut Pertanian Bogor, Bogor 16144
2Herbarium Bogoriense, Pusat Penelitian Biologi, Lembaga Ilmu Pengetahuan Indonesia, Bogor 16002

Thirteen species from three genera of Glomales fungi were identified from four cultivars of durian (Durio zibethinus Murr.) rhizosphere. They were Acaulospora foveata, A. longula, A. scrobiculata, A. tuberculata, Glomus aggregatum, G. albidum, G. etunicatum, G. fasciculatum, G. lacteum, G. rubiforme, G. versiforme, Glomus sp. 1, and Scutellospora calospora. Roots observation showed that durian is a mycotrophic plant. All arbuscular mycorrhizal symbiosis structures were observed in durian roots.

Key words: arbuscular mycorrhizal fungi (AMF), Durio zibethinus Murr., arbuscular mycorrhizal symbiosis structures

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Mikoriza Durian di Bogor dan Sekitarnya

Mycorrhiza of Durio in Bogor Area

CHAIRANI1, AGUSTIN WYDIA GUNAWAN1 & KARTINI KRAMADIBRATA2

1Jurusan Biologi, FMIPA, Institut Pertanian Bogor, Bogor 16144
2Herbarium Bogoriense, Pusat Penelitian Biologi, Lembaga Ilmu Pengetahuan Indonesia, Bogor 16002

Thirteen species from three genera of Glomales fungi were identified from four cultivars of durian (Durio zibethinus Murr.) rhizosphere. They were Acaulospora foveata, A. longula, A. scrobiculata, A. tuberculata, Glomus aggregatum, G. albidum, G. etunicatum, G. fasciculatum, G. lacteum, G. rubiforme, G. versiforme, Glomus sp. 1, and Scutellospora calospora. Roots observation showed that durian is a mycotrophic plant. All arbuscular mycorrhizal symbiosis structures were observed in durian roots.

Key words: arbuscular mycorrhizal fungi (AMF), Durio zibethinus Murr., arbuscular mycorrhizal symbiosis structures

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Isolasi dan Karakterisasi ?-amilase Ekstraseluler dari Kapang Asal Limbah Cair Tapioka

Isolation and Characterization of Extracellular ?-amylases Producing Moulds Isolated from Cassava Starch Liquid Waste

DEZI HANDAYANI, NISA RACHMANIA MUBARIK & SRI LISTIYOWATI

Jurusan Biologi, FMIPA, Institut Pertanian Bogor, Bogor 16144

A total of 25 moulds have been isolated from cassava starch liquid waste on the medium containing starch at pH 3.0 to 6.0. The isolates had amylase indexes from 0.03 to 2.50. Two isolates had the highest index at pH media 4.0 (0.84) and at 6.0 (2.5) were identified as Penicillium sp. strain K12 and Aspergillus versicolor 3a1 respectively. The optimal amylases production of Penicillium sp. strain K12 and A. versicolor 3a1 was obtained after 4 and 3 days of incubation at 30oC. The optimum of amylase activity from Penicillium sp. strain K12 was at 50oC and pH 5.0, while those from A. versicolor 3a1 was at 30oC and pH 6.0 respectively. The amylase activities of Penicillium sp. strain K12 increased after the addition of 10 mM of each Ca2+and Fe2+, while the amylase A. versicolor 3a1 increased after the addition of 10 mM Ca2+. Amylase activities of both isolate decreased after the addition of ethylenediaminetetraacetic acid at the concentration of 1 mM and 10 mM.

Key words: isolation ?-amylase, characterization, Penicillium, Aspergillus versicolor

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Ekspresi Gen inaZ pada Vibrio sp. untuk Memantau Pelekatan Bakteri pada Larva Udang

Expression of inaZ Gene in Vibrio sp. to Monitor Bacterial Adherence on Shrimp Larvae

YUSMINAH HALA1 & ANTONIUS SUWANTO1,2

1Jurusan Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144
2/sup>Seameo-Biotrop, Jalan Raya Tajur Km. 6, Kotak Pos 116, Bogor 16720

Expression of ice nucleation gene encoded by inaZ in several Vibrio sp. was investigated to understand the ability of the gene as a reporter for adherence and colonization of these presumably pathogenic bacteria in shrimp larvae. Conjugation was used to transfer a broad host range plasmid carrying ice nucleation gene which was constructed by inserting 4.5 kb EcoRI-PstI fragment containing inaZ gene from pIce1.9 into EcoRI-PstI sites in pRK415. The ice nucleation activity could be detected at (-9) – (-4) oC. A 10-day survival study of Vibrio sp. ice+ in 2% (wt/vol) NaCl and yeast extract showed that the bacterial population increased at day 3 and day 5 after inoculation and decreased at the following days. Meanwhile, ice nucleation activity decreased at day 3 and day 6 but still detectable until day 10. A 7-day adherence study indicated that ice nucleation activity of Vibrio sp. ice+ decreased at day 2 and become undetected at day 3. However the bacterial cell count of Vibrio sp. ice+ increased from day 2. The results suggested that the plasmid carrying ice nucleation gene was quite stable but the low expression of ice nucleation activity might be due to the unstability of InaZ protein. Neither Vibrio sp. ice+ nor ice- colonized the surviving larvae were observed after day 5 of incubation. It seems that they colonized the dead larvae in spite of the lack of observable ice nucleation activity.

Key words: inaZ gene, vibrio sp., bacterial adherence

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Deteksi Gen Cry1A Bacillus thuringiensis Berliner dengan Teknik PCR dan Toksisitasnya terhadap Ostrinia furnacalis Guenee

Detection of Cry1A Gene of Bacillus thuringiensis Berliner using PCR and its toxicity against Ostrinia furnacalis Guenee

BAHAGIAWATI1, HABIB RIJZAANI1 & NOVITA RIANI SIMANJUNTAK1

1Balai Penelitian Bioteknologi dan Sumberdaya Genetik Pertanian, Jalan Tentara Pelajar No. 3A, Bogor 16114
2/sup>Departemen Biologi, FMIPA, Universitas Indonesia, Kampus Depok, Depok 16424

The objectives of this experiment were to detect the presence of cry1A sequences from local Bacillus thuringiensis isolates multiplied by Lep1A and Lep1B primers using PCR technique and to determine their toxicity against Ostrinia furnacalis, maize stemborer. From 15 tested isolates, 11 of them gave PCR product. Among those 11 only 7 isolates, namely Lam 864, Jtg 2151, C 522, G 631, C 342, C 512, and C 536, showed a single, 490 bp DNA band, which is the expected size of PCR product using Lep1A and Lep1B primers. These isolates showed potentially high toxicity against maize stemborer. Four isolates, namely Ser 455, Cib 361, Lam 854, and Ser 554, showed the 490 bp band with a few additional bands, and had lower levels of toxicity to the tested insect. One isolate, C 423, did not give any PCR product but interestingly indicated a relatively high level of toxicity against the insect.

Key words: Bacillus thuringiensis, cry1, Ostrinia furnacalis, PCR

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Identifikasi Bakteri Penghasil Mananase serta Karakterisasi Enzimnya

Identification of Mannanase Producing Bacteria and Characterization of the Enzyme

DYANNE DOROTHY AURORA, YULIN LESTARI & ANJA MERYANDINI?

Jurusan Biologi, FMIPA, Institut Pertanian Bogor, Bogor 16144

Mannanase is a mannan degrading enzyme which is produced by microorganisms included bacteria. This enzyme can be used in many industrial processes as well as for improving the quality of animal feeds. The aim of this research was to isolate mannanolytic bacteria from copra soil sample which came from Pasaman, West Sumatra and characterize their enzymes. Screening was carried out using agar plates containing mannan stained with congo-red. Four isolates showed high mannanolytic index. DYP2, which was the best from the four bacteria was identified by 16S r-RNA sequence as Bacillus pumilus. DYP 2 produced optimum activity (0.079 U/ml) after 24 hour cultivation in the substrat containing gum locust bean. At an optimum activity, the protein concentration was 0.4 mg/ml and the specific activity was 0.2 U/mg. Mannanase of Bacillus pumilus DYP2 reached the optimum temperature at 80oC and the highest activity at pH 3.00. In the optimum condition the activity was stable for 5 days.

Key words: Bacillus pumilus, mannanase

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Coronavirus dan Sindroma Pernafasan Akut Berat

Coronavirus and Severe Acute Respiratory Syndrome

FERA IBRAHIM? & T. MIRAWATI SUDIRO

Bagian Mikrobiologi, Fakultas Kedokteran, Universitas Indonesia, Jalan Pegangsaan Timur No. 16, Jakarta 10320

Since first reported in July 2003, severe acute respiratory syndrome (SARS) has evoked a global alertness, and international collaboration among laboratories and countries. Until 12 July 2003, 8442 suspected and probable cases and 812 death has been reported. Through extensive studies, a new coronavirus, i.e. SARS virus was found to be the agent causing SARS. SARS virus is distinct from the other known members of coronavirus. SARS virus is readily cultivated in cell culture, SARS virus genomic sequence in various part of its genes showed that it belongs to a new group of coronavirus. Various techniques to detect SARS virus have been developed, although it’s not yet to the level of expected sensitivity and specificity.

Key words: SARS, Coronavirus, SARS-CoV

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Seleksi Bakteri Asam Laktat Indigenus sebagai Galur Probiotik dengan Kemampuan Menurunkan Kolesterol

Selection of Indigenous Lactic Acid Bacteria as Probiotics for Lowering Serum Cholesterol Levels

NETTY KUSUMAWATI1, BETTY SRI LAKSMI JENIE1, SISWA SETYAHADI2 & RATIH DEWANTI-HARIYADI1

1Fateta, Institut Pertanian Bogor, Kampus Darmaga, Bogor 16680
2Pusat Pengkajian dan Penerapan Teknologi Bioindustri, BPPT Gedung II Lt. 15,
Jalan M.H. Thamrin No. 8, Jakarta 10340

Eighteen strains of indigenous lactic acid bacteria isolated from Indonesian traditional fermented foods were tested for their potency as probiotic culture including survival in acid and bile as well as ability to assimilate cholesterol in vitrous. Three strains namely Lactobacillus plantarum sa28k, L. acidophilus FNCC116 and L. casei FNCC262 showed the highest ability to assimilate cholesterol and had good acid and bile resistance. These strains were further investigated for hypocholesterolemic effect in Sprague-Dawley Rats. Rats were fed with stock diet and drinking milk containing one of four treatments i.e. (i) non fermented milk (control), (ii) milk fermented by L. plantarum sa28k (F1), (iii) milk fermented by L. acidophilus FNCC116 (F2), and (iv) milk fermented by L. casei FNCC262 (F3). After 30 days, rats received the fermented milk had lower (P<0.05) serum cholesterol levels than did the non fermented milk rats. Serum cholesterol levels for control, F1, F2, and F3 rats were 67.5 mg/dl; 48.4 mg/dl; 51.0 mg/dl and 52.67 mg/dl respectively. Results indicate that indigenous strains of lactic acid bacteria isolated from Indonesian fermented foods can be considered as probiotic strains that have beneficial effect in reducing serum cholesterol levels.

¬¬ Key words: lactic acid bacteria, probiotic, cholesterol

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Produksi Massal Sel Rhizobium dengan Teknologi Bioproses

Mass Production of Rhizobium Using Bioprocess Technology

RASTI SARASWATI1, KHASWAR SYAMSU2, DWI NINGSIH SUSILOWATI1, BADRIYATUL LAILA2 & RENGGANIS SANTIKA ANDHAYANI2

1Balai Penelitian Bioteknologi Tanaman Pangan dan Sumberdaya Genetik Pertanian, Jalan Tentara Pelajar 3A, Bogor 16111
2Jurusan Teknologi Industri Pertanian, Institut Pertanian Bogor, Kampus Darmaga, Bogor 16680

Mass Production of Rhizobium Using Bioprocess Technology. Efficient mass production of Rhizobium inoculant has been studied by applying bioprocess technology under laboratory and pilot scale. The mass production of Rhizobium strain RIFCB1 … RIFCB7 has been optimized by batch system under pilot scale with maximum population of 2.5 x 109 sel/l after 48 h and maximum specific growth rate of 0.129 h-1 for 0-24 h, while under fed batch system the highest cell number was 2.3 x 109 sel/l after 24 h cultivation with maximum specific growth rate of 0.135 h-1 for 0-24 h. The mass production were decreased after 24 h and increased at 78 h to the maximum peak at 102 h with cell number of 1.7 x 109 cell/l. Fed batch system is much better than batch system due to the continuation of mass production. The addition of fresh medium could increase the cell number.

Key words: Rhizobium, bioproses

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Isolasi dan Identifikasi Mikroorganisme Termofil Isolat Kawah Wayang

Isolation and Identification of Thermophilic Microorganism from Wayang Crater

INDRAJAYA, FIDA MADAYANTI WARGANEGARA & AKHMALOKA*

Departemen Kimia, FMIPA, Institut Teknologi Bandung, Jalan Ganesha 10, Bandung 40132

Research on thermophilic microorganism has extensively been carried out, in this report we presented the isolation and identification of thermophilic microorganism from geothermal region around Bandung. Isolation had been performed from Wayang crater that has extreme environment with pH 1-2 and temperature 75-92oC. The microorganism was cultivated at 68oC in the modified Thermus medium by addition of mineral supplements. The 16S rRNA gene from one of the isolate was amplified by universal primers (27F and 1492R) resulting 1.5 kb DNA fragment. The fragment was cloned and sequenced. The phylogenetic analysis of the sequences suggested that the Wayang isolate is close to Geobacillus thermoleovorans.

Key words: thermophilic microorganism, 16S rRNA, isolation, identification, phylogenetic

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Isolasi dan Identifikasi Mikroorganisme Termofil Isolat Kawah Wayang

Isolation and Identification of Thermophilic Microorganism from Wayang Crater

INDRAJAYA, FIDA MADAYANTI WARGANEGARA & AKHMALOKA*

Departemen Kimia, FMIPA, Institut Teknologi Bandung, Jalan Ganesha 10, Bandung 40132

Research on thermophilic microorganism has extensively been carried out, in this report we presented the isolation and identification of thermophilic microorganism from geothermal region around Bandung. Isolation had been performed from Wayang crater that has extreme environment with pH 1-2 and temperature 75-92oC. The microorganism was cultivated at 68oC in the modified Thermus medium by addition of mineral supplements. The 16S rRNA gene from one of the isolate was amplified by universal primers (27F and 1492R) resulting 1.5 kb DNA fragment. The fragment was cloned and sequenced. The phylogenetic analysis of the sequences suggested that the Wayang isolate is close to Geobacillus thermoleovorans.

Key words: thermophilic microorganism, 16S rRNA, isolation, identification, phylogenetic

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Karakterisasi Protease Ekstraseluler

Clostridium bifermentans R14-1-b Characterization of Extracellular Protease from Clostridium bifermentans R14-1-b

JULIA ENGGEL1, ANJA MERYANDINI1 & LILY NATALIA2

1Departemen Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144
2Balai Penelitian Veteriner, Jalan Martadinata No. 30, Bogor 16114

Clostridium are anaerobic bacteria, Gram positif, rods shaped and form endospores. This bacteria are commonly found in soil, marine and fresh sediments, in the intestinal tract of man and animals. Samples were collected from intestinal contents of cattle, buffaloes and sheep from a slaughter house in Bogor, fresh sediments and soil mangrove from West Nusa Tenggara. From these 44 samples, 72 isolates Clostridium were screened for their ability to produce protease. The highest activivity was recorded from an isolate identified as C. bifermentans R 14-1-b and has a proteolitic index of 9.0. This bacteria was isolated from intestinal contents of sheep from slaughter house Bogor. C. bifermentans R 14-1-b showed maximum protease activity after 21 hours of cultivation in liquid media. Protease of C. bifermentans R 14-1-b displayed maximum activity at pH 7.5 and 50 oC with casein as substrate. Ions such as Cu2+, Co2+, and Zn2+ (5 mM) inhibited protease activity, whereas Zn2+ (5 mM) showed the strongest inhibition (87.50%). The protease was activated by Fe2+ ion at concentration 5 mM (322.50%). Mg2+ and Ca 2+ ions (5 mM) increase the protease activity slightly. The presence of chelating agent, such as Na2EDTA (1 mM and 5 mM) demonstrated the inhibitory effect of this enzyme, whereas Na2EDTA at concentration 5 mM showed the strongest inhibition (91.68%).

Key words: Clostridium, protease

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Karakterisasi Protease Ekstraseluler

Clostridium bifermentans R14-1-b Characterization of Extracellular Protease from Clostridium bifermentans R14-1-b

JULIA ENGGEL1, ANJA MERYANDINI1 & LILY NATALIA2

1Departemen Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144
2Balai Penelitian Veteriner, Jalan Martadinata No. 30, Bogor 16114

Clostridium are anaerobic bacteria, Gram positif, rods shaped and form endospores. This bacteria are commonly found in soil, marine and fresh sediments, in the intestinal tract of man and animals. Samples were collected from intestinal contents of cattle, buffaloes and sheep from a slaughter house in Bogor, fresh sediments and soil mangrove from West Nusa Tenggara. From these 44 samples, 72 isolates Clostridium were screened for their ability to produce protease. The highest activivity was recorded from an isolate identified as C. bifermentans R 14-1-b and has a proteolitic index of 9.0. This bacteria was isolated from intestinal contents of sheep from slaughter house Bogor. C. bifermentans R 14-1-b showed maximum protease activity after 21 hours of cultivation in liquid media. Protease of C. bifermentans R 14-1-b displayed maximum activity at pH 7.5 and 50 oC with casein as substrate. Ions such as Cu2+, Co2+, and Zn2+ (5 mM) inhibited protease activity, whereas Zn2+ (5 mM) showed the strongest inhibition (87.50%). The protease was activated by Fe2+ ion at concentration 5 mM (322.50%). Mg2+ and Ca 2+ ions (5 mM) increase the protease activity slightly. The presence of chelating agent, such as Na2EDTA (1 mM and 5 mM) demonstrated the inhibitory effect of this enzyme, whereas Na2EDTA at concentration 5 mM showed the strongest inhibition (91.68%).

Key words: Clostridium, protease

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Kemampuan Rizobia Bangkuang dari Berbagai Daerah di Indonesia dalam Menambat Nitrogen dan Meningkatkan Pertumbuhan Bangkuang

The Ability of Rhizobia Isolated from Yam Bean from Different Regions of Indonesia to Fix AtmosphericNitrogen and Increase the Growth of Yam Bean

ISWANDI ANAS & DINI MUFRIAH

Departemen Tanah, Faperta, Institut Pertanian Bogor, Kampus Darmaga, Bogor 16680

Fifty-four yam bean (Pachyrhizus erosus) cultivated soils collected from different regions of Indonesia were used as media to grow a local variety (Bogor) of yam bean. Root nodules of yam bean only were formed at 38 (70%) soils. Seventy rhizobia isolates were collected from root nodules and root of yam bean. Four (5.7%) out of 70 rhizobia isolates belonged to Bradyrhizobium spp. Twenty-six rhizobia isolates were further selected for their ability to increase the growth of yam bean and to fix atmospheric nitrogen. Twenty-two (84.6%) of rhizobia isolates tested were able to improve the growth (height, upper part biomass and root biomass) of local variety of yam bean and the rest four rhizobia isolates (15.4%) were not. The increase in yam bean growth was due to the increase in atmospheric nitrogen fixation by rhizobia isolates. Three rhizobia isolates (B-26/NS, B-27a/NS, and B-55/CJ) were able to increase plant growth (73% to 141%) and to fix atmospheric nitrogen (161% to 256%) higher than the treatment received 70 ppm N fertilizer (Kontrol + N).

Key words: Yam bean (Pachyrhizus erosus), rhizobia, nitrogen fixation, Acetylene Reduction Assay (ARA)

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Produksi dan Karakterisasi Parsial Protease Alkali Termostabil Bacillus thermoglucosidasius AF-01

Production and Partial Characterization of Thermostable Alkaline Protease from Bacillus thermoglucosidasius AF-01

ASRUL MUHAMAD FUAD1, RINI RAHMAWATI2 & NISA RACHMANIA MUBARIK2

1Pusat Penelitian Bioteknologi, Lembaga Ilmu Pengetahuan Indonesia, Jalan Raya Bogor Km. 46, Cibinong 16911
2Departemen Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144

Bacillus thermoglucosidasius AF-01, isolated from a traditional tofu waste at Ciampea – Bogor, produced a potentially thermostable alkaline protease. Maximum activity reached 132 U/ml using glucose or starch as carbon source in shake flask fermentation. Its activity was higher than those produced by reference isolates such as B. licheniformis (113 U/ml) or B. stearothermophilus (84 U/ml). Protease production using Durham’s media reached its maximal production at 124.6 U/ml after 27 hours of fermentation in a 1l fermentor under controlled pH and temperature (pH 9.0 and 45 oC). Protease from AF-01 might be a metalloprotease. Its activity was inhibited by the presence of EDTA but was not inhibited by other protease inhibitors such as PMSF or PCMB. It was revealed that some metal ions such as Cu2+, Ca2+, Mg2+, or Zn2+ increased the enzyme activity, whereas some others such as K+, Na+, dan Mn2+ decreased its activity. The activity was not affected by Co2+ ion and the presence of SDS up to 0.4% (w/v). However, enzyme activity was reduced in the presence of 0.5% (w/v) of SDS. The optimum pH and temperature of the enzyme activity were 9.0 and 80 oC respectively. Thermostability assay revealed that the enzyme was relatively stable at 80 oC up to 12 hours in which the enzyme retained 60% of its initial activity. Bacterial identification showed that the isolate AF-01 was belong to Bacillus. Further identification employing the BIOLOG system revealed that the isolate had similarity (0.269) to B. thermoglucosidasius.

Key words: Alkaline protease, thermostable, metalloprotease, Bacillus thermoglucosidasius

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Uji Kepekaan Helicobacter pylori Isolat Jakarta terhadap Berbagai Jenis Antibiotik

Susceptibility Test of Jakarta Isolates Helicobacter pylori Against Multiple Antibiotics

WIDYASARI KUMALA1, AZIZ RANI2 & BAMBANG HANDANA3

1Bagian Mikrobiologi, FK, Universitas Trisakti, Jalan Kyai Tapa No. 250, Jakarta 11440
2Bagian Penyakit Dalam Sub Bagian Gastroenterologi, FK, Universitas Indonesia, Jalan Diponegoro No. 71, Jakarta 10430
3Bagian Endoskopi Rumah Sakit Graha Medika, Jalan Raya Perjuangan Kav. 8, Jakarta 11530

Biopsies were done to eighty patients visiting Cipto Mangunkusumo National Centre Hospital and four private hospitals in Jakarta. Susceptibility test had been done to sixty isolates of Helicobacter pylori (H. pylori) which were taken from gastric mucous biopsy. Susceptibility test had been done to all of the isolates using Kirby Bauer method, which is recommended by National Committee for Clinical Laboratory Standards (NCCLS). The results showed that among the sixty isolates tested, there were 98% sensitivity to gatifloxacin, 95% to ciprofloxacin, 80% to amoxcycillin, 68% to clarithromycin, 63% to chloramphenicol, 60% to tetracyclin 58% to erithromycin, and 53% to ampicillin. There was no sensitivity to metronidazol. It appears that many H.pylori isolates begin to become resistant to antibiotics, especially to standardized used medical antibiotics. On the other hand quinolon group shows a satisfying high sensitivity, which can be used for further medical treatment and eradication of H. pylori.

Key words: Helicobacter pylori, susceptibility test, antibiotics

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Pembentukan Magnetosom pada Bakteri

Magnetosome Formation in Bacteria

ARIS TRI WAHYUDI

Departemen Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144
Tel./Fax. +62-251-345011, E-mail: aristri2003@yahoo.com

Magnetic bacteria orient and navigate along geomagnetic field lines and are widely distributed in freshwater and marine habitat. The ability of these bacteria to respond to magnetic fields is based on the presence of intracellular magnetosome, i.e. membrane-bound magnetic particle of either magnetite (Fe3O4) or greigite (Fe3S4). The magnetosome formation is achieved by tightly controlled processes which involves the accumulation of iron and deposition of the mineral particles at a specific location in the cell. Biomineralization of magnetosome may involve intricate processes, but the exact mechanisms is still poorly understood. This review focuses on the current knowledge about biochemical, physiological as well as molecular biological aspects of the biomineralization process of magnetic bacteria, especially in the magnetosome formation.

Key words: Magnetic bacteria, magnetosome, biomineralization

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Potensi Isolat Bakteri Penghasil IAA dalam Peningkatan Pertumbuhan Kecambah Kacang Hijau pada Kondisi Hidroponik

The Potency of IAA Producing Bacteria Isolates on Promotion The Growth of Mungbean Sprout in Hydroponic Condition

I NYOMAN P. ARYANTHA*, DIAN P. LESTARI & NURMI PURI DWI PANGESTI

Kelompok Penelitian dan Pengembangan Ilmu Hayati, LPPM Institut Teknologi Bandung,
Gedung Litbang ITB Lt. VI, Jalan Ganesha 10, Bandung 40132

Indole-3-acetic acid (IAA) is a key hormone for various aspects of plant growth and development. Liquid and powder products of five IAA producing bacteria (D2, D3 from bacilli group and KB, LE, LC from actinomycetes group) were investigated in semi in vivo assay towards mungbean (Vigna radiata) growth. Liquid fermentation product without cell separation was diluted 20, 40, and 60 times in sterilized water and dried powder was suspended in four concentrations i.e 0.01, 0.10, 1.00, and 3.00 g per 100 ml of water. Biological assay was conducted towards 3-day-old mungbean seedlings with hydroponic method in 20 ml tubes at room temperature and light intensity of 40 lux. The length of seedlings and root branching were assessed over 4 days. The data were analyzed by ANOVA. The powder product of KB at concentration rate of 0.01 g/100 ml (IAA = 0.021 ?g/ml) gave the highest seedling length (28.1 cm) that was significantly different (P < 0.05) compared with other treatments and control. For root branching, the liquid product of LC with 20 times dilution (IAA = 1.82 ?g/ml) gave the highest number (24.25) of branches that was significantly different (P < 0.05) compared with other treatments and control. These results indicate that these IAA-producing bacteria are potential to be used for promoting the growth of mungbean plant.

Key words: indole-3-acetic acid (IAA), Vigna radiata, Bacillus, Actinomycetes, soil bacteria, microbial phytohormone

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Pemanfaatan Lumpur Instalasi Pengolahan Air Limbah: Studi Pendahuluan terhadap Pertumbuhan Vegetatif Jagung Manis dan Mikrob Tanah

Utilization of Sludge from Waste Water Treatment: Preliminary Study on the Vegetative Growth of Sweet Corn and Soil Microbes

REGINAWANTI HINDERSAH1, MARTHIN KALAY2 & BARTI SETIANI MUNTALIF3

1Jurusan Tanah, Faperta, Universitas Padjadjaran, Jalan Raya Jatinangor Km. 21, Bandung 40600
2Jurusan Budi Daya Pertanian, Faperta, Universitas Pattimura, Kampus Poka, Ambon 97233
3Departemen Teknik Lingkungan, Fakultas Teknik Sipil dan Perencanaan, Institut Teknologi Bandung, Jalan Ganesha 10, Bandung 40132

Waste water treatment plant in Bandung faced a problem of dry sludge accumulation. This sludge contains some plant nutricents. However, its utilization as media growth for crops production is limited by high concentration of heavy metals (Pb and Cd). Preliminary study has been carried out to asses its influence on the vegetative growth of sweet corn, the growth of total soil bacteria and N2 fixing bacteria Azotobacter sp., and the colonization of vesicular-arbuscular mycorrhiza (VAM). The pot experiment demonstrated that adding dry sludge until 75% increased sweet corn vegetative growth, however, the 50% addition increased the soil concentration of Pb and Cd more than Inceptisols. The soil condition depresed the growth of either total soil bacteria or Azotobacter sp., and decreased the ability of soil fungi to form VAM.

Key words: dry sludge, sweet corn, soil microbes

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Isolasi dan Uji Kepekaan Isolat Klinis ORSA dan nonORSA terhadap Vankomisin dan Antibiotik Lainnya

Isolation and Susceptibility Tests Clinical Isolates of ORSA and nonORSA to Vancomycin and Other Antibibitics

HERA NOVIANA

Bagian Mikrobiologi, Fakultas Kedokteran, Unika Atma Jaya, Jalan Pluit Raya No. 2, Jakarta 14440 Tel. +62-21-6694366 pes 254, Fax. +62-21-6606123, E-mail: hnoviana@mail.com

Oxacillin-resistant Staphylococcus aureus (ORSA) or methicillin-resistant Staphylococcus aureus (MRSA) has become an important nosocomial pathogen in many countries. Vancomycin is currently the treatment of choice for serious infections caused by ORSA. However, with the emergence of these resistant isolates to vancomycin and other glycopeptides underscores the need other drugs that can provide an alternative antimicrobial agents for treatment of multi-drug-resistant S. aureus. The aim of the research is to monitor the effectiveness of Vancomycin and other antibiotics for infection caused by S. aureus. During May 18, 2000 to September 22, 2003 specimens were collected from patients at Atmajaya Hospital. The bacterial isolates were identified with biochemical reaction using Microbact and susceptibility tests were performed by a standard NCCLS method. Sixty-one S. aureus were isolated from many specimen and restricted into several categories: (i) ORSA (29 strains), the group included ORSA-VRSA (26 strains) and ORSA-VSSA (3 strains); (ii) nonORSA, the group included nonORSA-VRSA (30 strains) and nonORSA-VSSA (2 strains). ORSA-VRSA were multi-resistant to antibiotics. However, only meropenem was the choice for the infection caused by these group of ORSA-VRSA bacteria. ORSA-VSSA also were multi-resistant to antibiotics and vancomycin was sensitively intermediate instead. Both nonORSA and VRSA, as well as VSSA, are multi-sensitive to antibiotics especially to cephalosporin. In conclusion, if S. aureus have already resistant to oxacillin and vancomycin (ORSA-VRSA), they also will have developed other resistance to other antimicrobial drugs, so it will more difficult to treat infection caused by ORSA-VRSA.

Key words: ORSA (oxacillin-resistant S. aureus), VRSA (vancomycin-resistant S. aureus), VSSA (vancomycin-sensitive S. aureus)

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Pectinesterase Synthesis by Aspergillus niger Using Sugar Alcohol and Organic Acid Substrates

TRIWIBOWO YUWONO* & KOMARYATI DIAT ANGGERAWATI

Laboratory of Microbiology, Faculty of Agriculture, Gadjah Mada University, Bulaksumur, Yogyakarta 55281

A study has been conducted to determine the mode of pectinesterase synthesis in Aspergillus niger using non-carbohydrate carbon sources. The carbon sources used were sugar alcohol (mannitol and sorbitol) and organic acids (fumaric acid, citric acid, and malonic acid). The study demonstrated that pectinesterase was not synthesised when mannitol and sorbitol were used as carbon sources, suggesting that the mode of pectinesterase synthesis in A. niger is inductive. This was evidenced by the lack of methanol as a result of pectin hidrolysis by the filtrates following HPLC analysis. The alcohol substrates, however, were utilisable as carbon sources for growth. Organic acids, on the other hand, were used as carbon sources for growth as well as for inducing the synthesis of pectinesterase. The level of pectinesterase synthesis using organic acids as carbon sources was lower than the level of synthesis using pectin.

Key words: Aspergillus niger, pectinesterase, sugar alcohol, organic acids

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Kemampuan Bakteri Galur Lokal Tunggal dan Galur Campuran untuk Merombak Metidation

The Capacity of a Local Isolate and the Mixed Strains of Bacteria for Methidation Degradation

EDDY JUSUF

Pusat Penelitian Bioteknologi, Lembaga Ilmu Pengetahuan Indonesia, Jalan Raya Bogor Km. 46, Cibinong 16911
Tel. +62-21-8754587, Fax. +62-21-8754588, E-mail: eddy-jusuf@indo.net.id

The work was performed for testing the potency of a local isolate to degrade methidation, for bioremediation of the polluted soil. To evaluate its capacity, the mixed cultures of five bacterial strains obtained from culture collection of UPLB: Pseudomonas diminuta, P. putida strain 1503, P. putida strain 1504, P. putida strain 1506, and Bacillus cereus strain 1337 was used. Inoculants of single and mixed culture were made in composted manure and introduced into 4 kg of agricultural soil from Cibinong area containing 1200 ppm of methidation and incubated for 21 days. Analysis of the methidation residu from the treated soil was performed using HPLC with UV detector at 254 nm wavelength. The result showed that indigenous bacteria detected have played an important role in the degradation process by decreasing of 25.1% methidation in non-inoculated soil. Natural soil inoculated by the mixed cultures showed higher capacity by decreasing 50.6% of methidation whereas single local isolate 3b resulted 47.6%, but inoculation of sterilized soil by both types of cultures didn’t show any degradation activity. Either single isolate or mixed strains raised the capacity of methidation degradation in the soil so as indigenous microbes.

Key words: methidation degaradation , single and mixed cultures, soil bioremediation

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Produksi Senyawa Antimikrob dari Mobe dan Aplikasinya sebagai Bahan Pengawet Pangan

Production of Antimicrobial Compound from Mobe and Its Application as Food Preservatives

SEDARNAWATI YASNI1, YENNY ELISABETH2, ELVIRA SYAMSIR1 & ADOLF PARHUSIP3

1Departemen Teknologi Pangan dan Gizi, Fateta, Institut Pertanian Bogor, Kampus Darmaga, Bogor 16680
2Pusat Penelitian Kelapa Sawit, Jalan Brig. Jend. Katamso No. 5, Medan 20164
3Jurusan Teknologi Pertanian, Faperta, Unika St. Thomas, Jalan Setia Budi No. 479F Tj. Sari, Medan 20132

Mobe (Ficus sp.) is a spacy, that is popular in North Sumatera. The effect of mobe as antimicrobial on food pathogen bacteria was evaluated. Degree of maturity of the mobe fruit did not affect the antimicrobial activity against some tested bacteria and molds. Ethyl acetate and methanol extraction of mobe using maceration technique resulted in high antimicrobial activities. Ethyl acetate extract of mobe could inhibit the tested bacteria at lower concentration for less than or equal to 0.25% (w/v), except for B. cereus which required higher concentration of 0.75% (w/v). MIC value of the methanol extract was less than or equal to 0.75% w/v for all tested bacteria and molds, except for B. cereus which required higher MIC (Minimum Inhibitory Concentration) at concentration more than 0.75% w/v. The effectiveness of the application of mobe extract was determined by analyzing the TPC (Total Plate Count), TVN (Total Volatile Nitrogen), and TMA (Trimethylamine) contents of the fish fillet during storage. Application of the mobe extract on fish fillet by dipping was the most effective method while dispersing method was not effective and spraying was effective only for six hours of chill storage.

Key words: Mobe (Ficus sp.), extract, fraction, minimum inhibitory concentration (MIC), antimicrobial activity

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Deteksi Dini Infeksi Virus Nipah dengan Uji Enzyme Linked Immunoassay pada Babi di Indonesia

Early Detection of Nipah Virus Infection Using Enzyme Linked Immunoassay in Pigs in Indonesia

INDRAWATI SENDOW1, CHRIS MORRISSY2, TATTY SYAFRIATI1, DARMINTO1 & PETER DANIELS2

1Research Institute for Veterinary Science, P.O. Box. 151, Bogor
2Australian Animal Health Laboratory, Private Bag 3, Geelong, Victoria, Australia

Nipah is an exotic, zoonotic and contagious viral disease which infect pigs and humans. Using ELISA test, serological survey was conducted to gain the information of the presence of Nipah infection in pig in Indonesia. A total of 1471 pig sera collected from North Sumatera, Riau, North Sulawesi and Java were tested using inactivated Nipah antigen and the results indicated that no antibodies against Nipah virus was detected. In order to validate and to confirm the results recorded, 384 pig sera were send to Australian Animal Health Laboratory (AAHL) and were tested using serum neutralization test. Based on the same ELISA results and standardised ELISA techniques in both laboratories, it was concluded that Nipah ELISA test can be done at Balitvet laboratory and applied in Indonesia. The test is needed as a part of technological transfer, as well as to support the Government policy in order to anticipate the entering of Nipah virus infection.

Key words: virus nipah, serology, ELISA test, pigs

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Stabilitas Protease Logam Netral Mikrob Mirip Termolisin terhadap Panas

Thermostabily of Thermolysin Like Microbial Neutral Metaloprotease

BUDIASIH WAHYUNTARI

Bidang Teknologi Biokatalis, Deputi Bidang Teknologi Agroindustri dan Bioteknologi, Badan Pengkajian dan
Penerapan Teknologi, BPP Teknologi, Jalan M.H. Thamrin 8, Gedung BPPT II, lantai 15, Jakarta 10340
Tel. +62-21-3169509/7560536, Fax. +62-21-3169510/7560536, E-mail: budiasih@webmail.bppt.go.id

Termolysin is a neutral metaloprotease produced by Bacillus thermoproteolyticus. The enzyme is made up of a single chain of polypeptides consists of 300-316 amino acid residues that forms two domains. N end domain (amino acid residue 1-157) is mainly in the form of ?-pleated sheet and C end domain (amino acid residues 155-316) is ? helix shape. Both domains are connected by ? helix chain (amino acid residues 136-152) where some of the residues are important for the catalytic action of the enzyme. All neutral metaloproteases have one zinc atom that bound to their active sites and 2-4 calcium ions in each enzyme molecule that play important role on their thermostability. C end domain hardly plays role on the enzyme thermostability, on the other hand, amino acid Phe63 and Pro69 in N domain which is on the surface region which exposed to solvent might undergo early unfolding. Unfolding protein chain leads to its autolysis which responsible to thermal inactivation of the enzyme.

Key words: neutral metaloprotease, thermolysin, thermostability

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Stabilitas Protease Logam Netral Mikrob Mirip Termolisin terhadap Panas

Thermostabily of Thermolysin Like Microbial Neutral Metaloprotease

BUDIASIH WAHYUNTARI

Bidang Teknologi Biokatalis, Deputi Bidang Teknologi Agroindustri dan Bioteknologi, Badan Pengkajian dan
Penerapan Teknologi, BPP Teknologi, Jalan M.H. Thamrin 8, Gedung BPPT II, lantai 15, Jakarta 10340
Tel. +62-21-3169509/7560536, Fax. +62-21-3169510/7560536, E-mail: budiasih@webmail.bppt.go.id

Termolysin is a neutral metaloprotease produced by Bacillus thermoproteolyticus. The enzyme is made up of a single chain of polypeptides consists of 300-316 amino acid residues that forms two domains. N end domain (amino acid residue 1-157) is mainly in the form of ?-pleated sheet and C end domain (amino acid residues 155-316) is ? helix shape. Both domains are connected by ? helix chain (amino acid residues 136-152) where some of the residues are important for the catalytic action of the enzyme. All neutral metaloproteases have one zinc atom that bound to their active sites and 2-4 calcium ions in each enzyme molecule that play important role on their thermostability. C end domain hardly plays role on the enzyme thermostability, on the other hand, amino acid Phe63 and Pro69 in N domain which is on the surface region which exposed to solvent might undergo early unfolding. Unfolding protein chain leads to its autolysis which responsible to thermal inactivation of the enzyme.

Key words: neutral metaloprotease, thermolysin, thermostability

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Potensi Isolat Bakteri dari Kepiting Batu untuk Menghasilkan Minyak Kelapa secara Fermentasi

Bacterial Isolates of Mud-Crab and Their Potential to Produce Coconut Oil

DWI SURYANTO*, SITI KHADIJAH NASUTION & YURNALIZA

Jurusan Biologi, FMIPA, Universitas Sumatera Utara, Jalan Bioteknologi No. 1, Padang Bulan, Medan 20155

Nine bacteria isolated from the mud-crab (Grapsus sp.) from Pancur Batu, Deli Serdang, North Sumatra were identified and tested to produce coconut oil fermentatively. The isolates were grown individually in coconut milk cream media. Isolate SKN06 was able to produce coconut oil comparable to Saccharomyces cerevisiae and fleshy grinded of mud-crab, with the oil yields were 38.5 ml, 40 ml, and 37.5 ml, respectively. This means that isolate SKN06 has similar potential in producing coconut oil fermentatively compared to that of S. cerevisiae. The fatty acid index and water contents of the oil ranged from 0.56-1.23/g oil and 0.07-0.42%, respectively.

Key words: mud-crab (Grapsus sp.), coconut oil, fermentation

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Populasi Kapang Pascapanen dan Kandungan Aflatoksin pada Produk Olahan Kacang Tanah

Population of Storage Mold and Aflatoxin Content of Processed Peanut Products

LILIEANNY1, OKKY SETYAWATI DHARMAPUTRA1,2 & ASMARINA SETYANINGSIH RAHAYU PUTRI2

1Departemen Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144
2SEAMEO Biotrop, Jalan Raya Tajur Km. 6, Kotak Pos 116, Bogor 16001

This study was a survey to observe moisture content, population of storage mold, and aflatoxin content of peanut processed products, together with physical quality of peanut kernels. A total of 88 samples of roasted peanuts with skin pod, flour-coated peanuts, roasted peanuts without skin pod, bumbu pecel (dry peanut sauce), and enting-enting gepuk (peanut sweet) were collected from several factories, supermarkets, and traditional markets in Bogor, Malang, Pati, and Yogyakarta. The moisture contents of roasted peanuts with skin pod, flour-coated peanuts, roasted peanuts without skin pod and enting-enting gepuk were about 3% those were lower than that of bumbu pecel which was 10.5%. The percentages of intact kernels, shrivelled and damaged kernels in roasted peanuts with skin pod and roasted peanuts without skin pod were 79.1, 13.1, 7.4% and 83.6, 4.5, 11.8%, respectively. The percentages of intact kernels and shrivelled kernels in flour-coated peanuts were 93.1 and 6.8%, respectively. Damaged kernels were not found in flour-coated peanuts. The most often isolated molds in processed peanut products were as follows: Penicillium citrinum in roasted peanuts with skin pod was 70.2% of the samples; Cladosporium cladosporioides in flour-coated peanuts was 63.6%; Aspergillus penicillioides, C. cladosporioides, and Pestalotiopsis guepinii in roasted peanuts without skin pod were 66.7, 66.7, and 66.7%, respectively; both A. flavus and Eurotium chevalieri in bumbu pecel were 50%, respectively; both A. flavus and P. citrinum in enting-enting gepuk were 100%, respectively. The mold population in roasted peanuts with skin pod, flour-coated peanuts, roasted peanuts without skin pod, bumbu pecel and enting-enting gepuk were 30, 26, 14, 8, and 48 cfu/g dry weight, respectively. Roasted peanuts with skin pod, flour-coated peanuts, bumbu pecel and enting-enting gepuk had aflatoxins B1 and B2; B1 and G1; B1, B2 and G1; and aflatoxins B1 and B2, respectively. Their total aflatoxins contents were 1.8, 5.2, 41.6, and 20.8 ppb, respectively. Aflatoxin was not detected in roasted peanuts without skin pod.

Key words: aflatoxin, mold, peanuts

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Karakterisasi Kitosanase Ekstraseluler dari Bakteri Asal Air dan Tanah di Kota Matsue, Jepang dan Dibandingkan dengan Kitosanase dari Matsuebacter chitosanotabidus 3001

Characterization of Extracellular Chitosanase of Bacteria from Water and Soil at Matsue City, Japan Compared with Matsuebacter chitosanotabidus 3001

MARIA ENDO MAHATA1, YUN CHOONG SOO2, ABDI DHARMA3, IRSAN RYANTO1, YOSE RIZAL1 & MAKOTO KAWAMUKAI2

1Program Studi Ilmu-Ilmu Pertanian Pemusatan Peternakan, Pascasarjana, Universitas Andalas, Kampus Unand Limau Manis, Padang 25163;
2Department of Biochemistry and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Japan 690-8504;
3Program Studi Ilmu Kimia, Pascasarjana, Universitas Andalas, Kampus Unand Limau Manis, Padang 25163

One hundred twenty isolates of bacteria as chitosanase producer were isolated from water and soil around Matsue city, Japan. Four isolates were analyzed for chitosanase activity, and 2 strains among them (97 and 99) have shown high chitosanase activity compared with Matsuebacter chitosanotabidus 3001, and then their enzymes were characterized. Chitosanases from 97 and 99 like M. chitosanotabidus 3001 were stable on temperature between 20-50 oC, and the chitosanase from 99 and M. chitosanotabidus 3001 showed optimum temperature on 50 oC while that from 97 on 40 oC. Optimum pH value of chitosanase from 97 and 99 were 4 and 5, respectively and from M. chitosanotabidus 3001 was observed at pH 7. Chitosanase from 99 and M. chitosanotabidus 3001 were stable on pH range between 4-8 and from 97 on pH between 4-9. The enzyme of strain 97 likes M. chitosanotabidus 3001 was a chitosanase that can digest only chitosan, while that from strain 99 can hidrolyze chitosan and colloidal chitin.

Key words: chitosanase, characterization, water and soil bacteria

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Identifikasi Bakteri Bukan Penghasil Asam Laktat yang Berasosiasi dengan Tempoyak (Durian Fermentasi)

Identification of Non-Lactic Acid Bacteria Associated with Tempoyak (Fermented Durian)

NETI YULIANA

Jurusan Teknologi Hasil Pertanian, Faperta, Universitas Lampung, Jalan Sumantri Brojonegoro 1, Bandar Lampung 35145; Tel. +62-721-747105, Fax. +62-721-783682, E-mail: netiyuliana@yahoo.com

The presence of Bacillus sp. as food contaminant in tempoyak (fermented durian) has been previously indicated, although none of the detail species was really known. Biochemical assay and microscopic observation, including scanning electron microscopy were employed to pinpoint the associated Bacillus. We identified for the first time that Bacillus megaterium may be associated with lactic acid bacteria and therefore, being involved in the fermentation process.

Key words: tempoyak, Bacillus megaterium

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Identifikasi Geminivirus yang Menginfeksi Tomat Berdasarkan pada Teknik Polymerase Chain Reaction-Restriction Fragment Length Polymorphism

Identification of Geminivirus Infecting Tomato Based on Polymerase Chain Reaction-Restriction Fragment Length Polymorphism

NOOR AIDAWATI1,3, SRI HENDRASTUTI HIDAYAT1, RUSMILAH SUSENO1, PURNAMA HIDAYAT1 & SRIANI SUJIPRIHATI2

1Departemen Proteksi Tanaman;
2Departemen Agronomi dan Holtikultura, Faperta, Institut Pertanian Bogor, Kampus Darmaga, Bogor 16680
3Jurusan Hama dan Penyakit Tumbuhan, Faperta, Universitas Lambung Mangkurat, Jalan Jenderal A. Yani Km. 36, Banjarbaru 70714

Geminivirus is an important virus that can cause severe damage on various crop. Infection of geminiviruses, mediated through its insect vector Bemisia tabaci Genn., has been reported to cause significant loss on tomato industry in South America. Availability of detection and identification method with high accuracy and sensitivity is very critical for early warning system of the geminivirus disease. This research is conducted to develop an identification method to differentiate isolates of geminivirus based on PCR-RFLP technique. Using specific primers, PAL 1v 1978 and PAR 1c 715, DNA of geminivirus was successfully amplified from Central Java (Sawangan and Boyolali), DIY (Kaliurang and Kulonprogo), and West Java (Barusireum). Amplified viral DNA with the size of ? 1.6 kb was then subjected to restriction enzyme (BamHI, EcoRI, HindIII, and PstI). Based on the restriction pattern, geminivirus isolates from Java can be compared. Isolates from Sawangan were similar with those from Kaliurang. Both isolates were different from those of Boyolali, Kulonprogo, and Barusireum, whereas the last three isolates were different from each other. Using NTSYS PC 2.1 it can be concluded that the geminivirus infecting tomato in Java can be differentiated into two groups.

Key words: geminivirus, PCR-RFLP

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Isolasi Virus Palyam dari Sapi Sentinel di Timor Barat, Indonesia

Isolation of a Palyam Group Viruses from Sentinel

INDRAWATI SENDOW

Bagian Virologi, Balai Penelitian Veteriner, P.O. Box. 151, Jalan R.E. Martadinata No. 30, Bogor 15114 Tel. +62-251-331048, E-mail: i.sendow@balitvet.org

Palyam is one of arboviral disease which infected pregnant cow and caused abortion. To obtain the Palyam virus isolate, a group of sentinel cattle was established at Kupang, West Timor. A weekly heparinised blood was collected from Vena jugular for viral isolation. A total of 486 heparinesed blood were inoculated into baby hamster kidney (BHK-21) monolayer in roller tubes. Three times blind passages in BHK-21 was carried out and cythopathic effect (CPE) was observed daily before considered as negative and discarded. When CPE was observed, the inoculum considered containing viral isolate. Six isolates produced CPE, and further identification using agar gel immunodiffusion test against Palyam antisera was conducted. The result indicated that two isolates reacted with Palyam antisera. The isolates were further confirmed at Onderstepoort Laboratory, South Africa.

Key words: Palyam, isolation, cattle, identification

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Optimasi Media Tumbuh untuk Peningkatan Aktivitas dan Isolasi Enzim Superoksida Dismutase dari Biomassa Spirulina platensis

Growth Medium Optimization to Increase Activity and Isolation of Superoxide Dismutase from Biomass of Spirulina platensis

TRI PANJI1,2, MARINI WIJAYANTI3, SUHARYANTO1 & MARIA BINTANG4

1Balai Penelitian Bioteknologi Perkebunan Indonesia, Jalan Taman Kencana No. 1, Bogor 16151
2Departemen Kimia, FMIPA, Universitas Pakuan, Jalan Pakuan P.O. Box. 452, Bogor 16161
3Sekolah Pascasarjana, Institut Pertanian Bogor, Kampus Darmaga, Bogor 16680
4Departemen Kimia, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144

Spirulina platensis is a blue-green microalga or cyanobacteria producing several bioactive compounds such as superoxide dismutase (SOD) which is important in pharmaceutical and cosmetic industries as an antioxidant for free radical scavenger. To produce SOD in S. platensis biomass efficiently, it is necessary to define an optimum medium composition consisting of mineral salts and organic complex from serum latex. The alga was cultured on medium containing latex serum with and without mineral suplementation, and on a synthetic medium of Aiba and Ogawa as a reference. The culture was illuminated with 20 W TL lamp at room temperature (28-32 oC) for two months. The growth was observed by measuring absorbance at ? 480 nm, while SOD activity was determinated by measuring red color of azo dye stuff at ? 540 nm equivalent to SOD inhibition of hydroxylamine oxidation. Research result showed that the best growth was obtained on serum latex medium, followed by serum latex with mineral supplementation, and synthetic medium. A maximum SOD activity in biomass was achieved in S. platensis culturing on serum latex medium illuminated with 20 W TL lamp at room temperature (28-32 oC) for 28-days period. The SOD activity of the microalga was the same as SOD activity in imported Spirulina biomass. The SOD from S. platensis was possibly Zn-SOD isoform type since it is quite sensitive against cyanide and its molecule contains higher concentration of Zn compared to that of Mn, Fe, and Cu. Sensitivity to cyanide was higher in the purified product compared to that in crude extract.

Key words: microalga, Spirulina platensis, serum latex, superoxide dismutase

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Tingkat Kedinian Infeksi Acaulospora tuberculata dan Gigaspora margarita pada Bibit Kelapa Sawit

Earliest Level of Acaulospora tuberculata and Gigaspora margarita Infection on Oil Palm Seedling

HAPPY WIDIASTUTI1,2, NAMPIAH SUKARNO1, LATIFAH KOSIM DARUSMAN1, DIDIEK HADJAR GOENADI2, SALLY SMITH3 & EDI GUHARDJA1

1FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144
2Balai Penelitian Bioteknologi Perkebunan Indonesia, Jalan Taman Kencana No. 1, Bogor 16151
3School of Earth and Environmental Sciences, The University of Adelaide, Australia

Earliest level of infection of Acaulospora tuberculata and Gigaspora margarita in oil palm Elaeis guineensis, Jacq seedling was studied. A glasshouse experiment was conducted using polybag (10 x 10 cm) containing sterilized sand as medium. Inoculation was conducted by inoculating sorghum before the oil palm seedling. Four treatments assessed were combination of AM fungi species (A. tuberculata and G. margarita) and elimination of endosperm (with or without) with four replications each. Endosperm elimination was conducted in oil palm with two leaves. Observation of AM fungal infection was done every week during 14 weeks. The result showed that A. tuberculata and G. margarita infection in oil palm seedling could be observed after seven week of inoculation. The infection process was shortened one week in the oil palm seedling inoculated with G. margarita eliminated endosperm in all type of root. Infection of AM fungi in secondary root was higher compared to those in primary root. AM fungal structure observed was apresorium, internally hyphae, coiled hyphae, and arbuscle. Vesicle was observed only in oil palm root inoculated with A. tuberculata.

Key words: Elaeis guineensis, earliest level of infection, Acaulospora tuberculata, Gigaspora margarita

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Karakterisasi Protease Ekstraseluler Clostridium litusburense Me1-3 Asal Danau Meraran dari Nusa Tenggara

Characterization of Extracellular Protease Produced by Clostridium Me1-3 from Meraran Lake in East Nusa Tenggara

ANJA MERYANDINI

Departemen Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144 Tel. +62-251-377169, E-mail: ameryandini@yahoo.com

Clostridium lituseburense Me1-3 isolated from Meraran lake in East Nusa Tenggara was identified based on its 16S rRNA sequence analysis. This isolate produced maximum protease activity after 21 hours of cultivation in liquid media. The protease displayed maximum hydrolitic activity at 30 oC and pH 7.5. Divalent ion Co2+ at 5 mM concentration increased enzyme activity to approximately 300%.

Key words: Clostridium lituseburense, protease

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Analisis Zimogram a-Amilase Mikrob Toleran Basa

Zymogram Analysis of a-Amylases from Alkaline Tolerant Microbes

NISA RACHMANIA MUBARIK

Departemen Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144
Tel. +62-251-345011, E-mail: riknisa@telkom.net

Amylolyticalkaline tolerant microbes identified as Aspergillus sydowii K10, A. versicolor L30, and Bacillus firmus KH.9.4, were isolated from tapioca waste liquid in Bogor, produced high a-amylase activity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the enzyme produced by each of the microbes yields several bands of proteins. Zymographic analysis showed that the enzyme of A. sydowii K10 had two bands of a-amylases, those from A. versicolor L30 and B. firmus KH.9.4 had one band, respectively. The molecular weight of a-amylases from A. sydowii K10 were 16.2 kD and 56.5 kD, while those from A. versicolor L30 and B. firmus KH.9.4 were 23.5 kD and 63.8 kD, respectively.

Key words: a-amylase, zymogram, alkaline tolerant, Aspergillus sydowii, Aspergillus versicolor, Bacillus firmus

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Stimulasi Glutamat terhadap Produksi Lovastatin oleh Aspergillus terreus

Glutamate Stimulation on the Production of Lovastatin by Aspergillus terreus

1Balai Pengkajian Bioteknologi, Badan Pengkajian dan Penerapan Teknologi, Gedung 630, Kawasan Puspiptek Serpong, Tangerang 15310; 2Pusat Pengkajian dan Penerapan Teknologi Bioindustri, Badan Pengkajian dan Penerapan Teknologi, Jalan M.H. Thamrin, Jakarta 10320; 3Departemen Teknologi Industri Pertanian, Fateta, Institut Pertanian Bogor, Kampus Darmaga, Bogor 16680

The influence of nitrogen sources for lovastatin production by Aspergillus terreus employing glucose-skim milk as a carbon source was studied. Nitrogen sources used in these experiments were KNO3, urea, soy flour, and monosodium glutamate. Experiments using all media combination were conducted in 250 ml Erlenmeyer flask, incubated at 280C under shaking at 330 rpm for nine days. The results showed that nitrogen sources had significant effect on lovastatin production by Aspergillus terreus. Highest lovastatin production (772 ppm) was achieved when monosodium glutamate at 10 g l-1used as nitrogen source, in combination with glucose 45 g l-1 and skim milk 20 g l-1.

Key words: lovastatin, nitrogen sources, Aspergillus terreus

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Deteksi Xanthomonas axonopodis pv. glycines pada Kedelai dengan Teknik ELISA

Detection of Xanthomonas axonopodis pv. glycines on Soybean Employing ELISA Technique

1Departemen Proteksi Tanaman, Faperta, Institut Pertanian Bogor, Kampus Darmaga, Bogor 16680
2Departemen Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144

Detection of Xanthomonas axonopodis pv. glycines, the causal of bacterial pustule disease in soybean, is usually conducted by close examination of disease symptoms and culturing the pathogen, which are laborius and relatively slow. The objective of this study was to produce and evaluate polyclonal antisera using ELISA technique for detection of X. axonopodis pv. glycines from isolated bacterial culture and diseased soybean leaves. A polyclonal antisera (designated as APYR32 and APJA4) was produced by immunizing intravenously a three month-old rabbit with inactivated cell of X. axonopodis pv. glycines YR32 and JA4 at concentration of 108 cell ml-1. The antiserum was tested for its sensitivity, specifity and aplicability to diagnose the presence of X. axonopodis pv. glycines on soybean employing inderect-ELISA technique. The results showed that indirect ELISA technigue was sensitive in detecting as few as 103 cell ml-1 at a titre of 1:2000. Polyclonal antisera YR32 reacted with all X. axonopodis pv. glycines isolates from culture with different both location and host variety, although it also showed some cross reactivity with X. campestris pv. campestris at 104 cell ml-1. This technique would be useful to detect bacterial pustule disease, since APYR32 only reacted with natural or artificially infected soybean leaves but not with healthy leaves or leaves infected Pseudomonas syringae pv. glycinea.

Key words: Xanthomonas axonopodis pv. glycines, polyclonal antisera, ELISA

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Perkembangan Spora Acaulospora foveata

The Development of Acaulospora foveata’s Spore

1Herbarium Bogoriense, Pusat Penelitian Biologi, Lembaga Ilmu Pengetahuan Indonesia, Bogor 16122 2Departemen Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144

The development of Acaulospora foveata from trap culture and monospecific culture which was inoculated with seedlings of Pueraria phaseoloides from iwul (Orania sylvicola) rhizosphere is presented. A. foveata was found mojarity associated with iwul which is a dominant species in Dungus Iwul Nature Reserve, a remnant lowland rainforest in West Java. The result from trap culture and monospecific culture showed the spore development of a young spore which was produced from the neck of the hyphal terminus up to a mature spore. Trap culture also showed the development of dual spores produced from the neck of a hyphal terminus, but it was not observed in the monospecific culture.

Key words: Acaulospora foveata, spore

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Analisis Fragmen Gen HA1 Virus Avian Influenza yang Menginfeksi Sebuah Breeding Farm pada Awal Tahun 2004 dan 2005 di Kabupaten Sukabumi

Analisys of HA1 Gene Fragment of Avian Influenza Virus that Infected Breeding Farm in Early 2004 and 2005 in Sukabumi District

Balai Penelitian Veteriner, Jalan R.E. Martadinata No. 30, Bogor 15114
Tel. +62-251-331048, Fax. +62-251-336425, E-mail: nlpdharmayanti@yahoo.com

In early 2004 and 2005, H5N1 influenza viruses emerged among chickens in Breeding Farm in Sukabumi. Breeding farm is one of area target to eradicate of AI in Indoensia. Therefore, we isolated the virus in the field and conducted the homology of the viruses that infected this breeding in fragment HA1 gene. Homology and phylogenetic analyses showed that isolate from breeding farm in 2004 (A/Chicken/West Java/Smi-BF/2004) has homology 92% and there were defferences if compared with isolete from 2005 that infected the breeding farm (A/Chicken/West Java/Smi-27M/2005).

Key words: HA1 gene fragment, avian influenza, breeding farm

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Konstruksi Pustaka Genom Magnetospirillum magneticum AMB-1 dan Penapisan Gen yang Terlibat dalam Sintesis Magnetosom

Construction of Genomic Library of Magnetospirillum magneticum AMB-1 and Screening of Genes Involved in Magnetosome Synthesis

Departemen Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144
Tel./Fax. +62-251-345011, E-mail: aristri2003@yahoo.com

Magnetospirillum magneticum AMB-1 is a fresh-water magnetic bacterium capable of synthesizing membrane-bound magnetic particle, called magnetosome, under microaerobic conditions. To screen and isolate genes involved in magnetosome synthesis, genomic library of M. magneticum AMB-1 was constructed using pJRD215 as a cosmid vector. The genomic DNA of AMB-1 was partially digested with Sau3AI and ligated into BamHI digestion of pJRD215. The ligated DNA was packaged into bacteriophage ??particle and subsequently infected into Escherichia coli NM554 as a host strain. A thousand and five colonies of NM554 library carrying the recombinant cosmids were obtained that grew on luria agar supplemented with kanamycin (25 ?g ml-1) and streptomycin (50 ?g ml-1). Screening of the library by colony hybridization revealed that the library carried all DNA sequences or genes involved in magnetosome synthesis used as probes. The average of the genome size inserted in the library was approximately 10-40 kb. These large fragments would facilitate complementation analysis of genes involved in magnetosome synthesis in this forming magnetic bacterium.

Key words: Magnetospirillum magneticum AMB-1, Genomic library, Cloning, Colony hybridization

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Kelimpahan Bakteri Filosfer pada Beberapa Sayuran Lalaban

The Abundance of Phyllosphere Bacteria Isolated from Some Indonesian Fresh Leafy Salad (Lalaban)

Departemen Biologi, FMIPA, Institut Pertanian Bogor, Jalan Raya Pajajaran, Bogor 16144

Several phyllosphere bacteria were investigated using plate count method. Pink-pigmented facultative methylotroph (PPFM), phyllosphere, streptomycin resistant, and endospore-forming bacteria were isolated and counted from four Indonesian lalaban such as poh-pohan, lettuce, basil, and bean sprout. This research shows the number of bacteria present on those leafy salads. The number of PPFM bacteria was approximately 8.75 x 102 to 6.62 x 104 cfu g-1. Total phyllosphere bacteria was 1.07 to 2.90 x 108 cfu g-1, while streptomycin resistant bacteria was 4.63 x 104 to 1.35 x 105 cfu g-1, and endospore-forming bacteria was 5.68 x 102 to 6.67 x 103 cfu g-1. The number of PPFM was relatively lower compared to the other group of bacteria analyzed. However, these phyllosphere bacteria might contribute to the quality of lalaban diet as source of pyrroloquinoline quinone (PQQ), a novel vitamin which can act as antistroke or antioxidant.

Key words: phyllosphere bacteria, PPFM, Indonesian leafy salad

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Preparasi Sel Kering Lactobacillus sp. Dad 13 dan Kestabilannya sebagai Bubuk robiotik

Preparation of Dried Cell of Lactobacillus sp. Dad 13 and It’s Stability as Probiotic Powder

SRI HARTATI1 & ENI HARMAYANI2

1Jurusan Teknologi Hasil Pertanian, Faperta, Universitas Veteran Bangun Nusantara
Jalan Letjend S. Humardani No. 1, Sukoharjo 57521
2Jurusan Teknologi Pangan dan Hasil Pertanian, Fateta, Universitas Gadjah Mada,
Jalan Sosio Yustisia, Bulaksumur, Yogyakarta 55281

Lactobacillus sp. Dad 13 is an indigenous bacterium isolated from buffalo milk fermented (“dadih�) which has potential properties to reduce cholesterol. For the developments of probiotic instant drinks, the application of dried Lactobacillus sp. Dad 13 had been studied. The purposes of the study were to prepare a dried Lactobacillus sp. Dad 13 and to study its properties to assimilate cholesterol and to deconjugate bile salts during storage. Preparation of dried cells was done by spray drying with various filler (skim, maltodextrin and rice flour). The dried cells were selected then stored at room temperature and 4-6oC using vacum and nonvacum packaging. The cell viability, cholesterol assimilation and bile salt deconjugation were analysed every week for two months. The resuls of the study indicated that the population cell of Lactobacillus in skim, maltodextrin and rice flour filler were not significantly different at the number of 1.76 x 1010, 6.3 x 1010, dan 2.62 x 1010 CFU g1 respectively. The highest yield of cell dried was obtained using maltodextrin. The cell viability and bile salt deconjugation of Lactobacillus in maltodextrin filter were stable after storage for two months in refrigerator temperature. The cholesterol assimilation of the dried cell was remain the same after one month storage at 4-6oC.

Key words: Lactobacillus sp., probiotic, dried cell, stability, cholesterol

=========================================================

Produksi Asam Hialuronat oleh Streptococcus zooepidemicus ATCC 39920

Production of Hyaluronic Acid by Streptococcus zooepidemicus ATCC 39920

ERLIZA NOOR1*, EFRIDA MARTIUS2, FAKHRINA FAHMA3 & ENDANG GUMBIRA SA’ID

1Departemen Teknologi Industri Pertanian, Fateta, Institut Pertanian Bogor, Kampus Darmaga, Bogor 16680
2Balai Pengkajian Bioteknologi, Badan Pengkajian dan Penerapan Teknologi,
Gedung Kawasan Puspiptek Serpong, Tangerang 15310
3Departemen Teknologi Industri, Universitas Negeri Sebelas Maret,
Jalan Ir. Sutami 36A, Surakarta 57126

Hyaluronic Acid (HA) produced for pharmaceuticals, medical, and cosmetic industries is a high molecular weight of linear polysaccharide. HA, the cementing substance in connective tissue can be extracted from rooster comb and umbilical cord. More recently, the HA production has been enhanced by microbial cultivation. The objective of this research was to optimize the HA production by using media containing different concentrations of Na2HPO4•2H2O; KH2PO4•7H2O; MgSO4•7H2O and amino acids. The effects of temperature (33, 37, and 40oC) and agitation were also observed in batch cultures. Result show that medium containing high concentration of Na2HPO4•2H2O and complete amino acids gave high viscosity. The HA production was relatively better in the agitated culture than that in without agitation. The highest production of 9.7 g l-1 was observed in the culture at 100 rpm and 37oC.

Key words: hyaluronic acid, Streptococcus zooepidemicus

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Perbaikan Pertumbuhan Bibit Sengon pada Tanah Mineral Masam dengan Inokulan Rhizobium

Improvement of Paraserianthes Seedling Growth on Acid Mineral Soils by Using Rhizobium Inocula

DENI ELFIATI1‡, ISWANDI ANAS1* & LUKMAN GUNARTO2

1Departemen Ilmu Tanah dan Sumberdaya Lahan, Faperta, Institut Pertanian Bogor
Kampus Darmaga, Bogor 16680
2Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian,
Jalan Tentara Pelajar No. 3A, Bogor 16111

The main problems using acid mineral soil for agriculture are low in fertility, high in exchangable Al and high P-retention. Soil acidity inhibits plant growth, Rhizobium growth and root nodule formation. Rhizobium were isolated from acid mineral soils and selected for their effectivities to increase the growth of Paraserianthes seedlings. The results of experiment showed that inoculation of Paraserianthes seedlings with acid tolerance Rhizobium improved the quality of the Paraserianthes seedling. The best improvement of Paraserianthes seedling growth planted on Ultisol and Inceptisol Darmaga soils was obtained through inoculation with Rhizobium isolates GR2-7 and GR3-4. The inoculation of the Paraserianthes seedling with these Rhizobium isolates significantly increased the height of Paraserianthes seedling by 67 and 64% on Ultisol and by 90 and 122% on Inceptisol, respectively. The diameter of the stamp were increased by 36 and 21% on Ultisol and by 37 and 39% on Inceptisol, respectively. The upperparts biomass of Paraserianthes seedlings were incresed by 167 and 130% on Ultisol. Inoculation of Rhizobium GR2-7 and GR3-4 increased the N-uptake by 353 and 258% on Ultisol and by 575 and 643% on Inceptisol, respectively.

Key words: acid mineral soil, Paraserianthes seedling, Rhizobium

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Peranan Kosubstrat terhadap Aktivitas Enzim dalam Metabolisme Xilosa untuk Produksi Xilitol oleh Candida shehatae WAY 08

Effect of Cosubstrates on the Key Enzymes in Xylose Metabolism for Xylitol Production by Candida shehatae WAY 08

WISNU ADI YULIANTO‡*, KAPTI RAHAYU KUSWANTO, TRANGGONO & RETNO INDRATI

Fakultas Teknologi Pertanian, Universitas Gadjah Mada, Jalan Sosio Yustisia, Bulaksumur, Yogyakarta 55281

The experiment investigated the influence of arabinose, glucose, galactose, and mannose, as a substrat and cosubstrate in xylose metabolism and xylitol production by Candida shehatae WAY 08. Activity of key enzymes involved in the xylose metabolism was measured under substrates containing 6% of each sugars and different ratio addition of 6% xylose with 1, 2, or 3% other sugars as cosubstrate. The enzyme activity on 6% xylose without cosubstrate was used as a control. The activity of xylose reductase (XR) and glucose-6-phosphate dehydrogenase (G6PDH) were induced by 1% of arabinose addition, while glucose, galactose, and mannose repressed the activities of both enzymes. The activity of xylitol dehidrogenase (XDH), acetaldehide dehydrogenase, and alcohol dehydrogenase were slightly repressed by cosubstrates. Specific growth rate of C. shehatae with hexose (glucose, galactose, and mannose) was higher than that of pentose (arabinose and xylose). Addition of 1% arabinosa to 6% xylose didn’t decrease specific growth rate, in fact it induced XR and G6PDH, however it slightly repressed XDH and produced optimum xylitol production. The ratio of XR:XDH:G6PDH in that condition was found to be 47:10:148.

Key words: Candida shehatae WAY 08, cosubstrates, xylose, xylitol

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Identifikasi Bakteri Diazotrof Endofit dari Tebu dengan Repetitive-PCR Sequence dan Sequencing 16S rDNA

Identification of Endophyte Diazotrophic Bacteria from Sugarcane by Using Repetitive-PCR Sequence and Sequencing of 16S rDNA

WIWIK E. WIDAYATI1*, LANGKAH SEMBIRING2 & J. SOEDARSONO3

1Pusat Penelitian Perkebunan Gula Indonesia, Jalan Pahlawan 25, Pasuruan 67126
2Fakultas Biologi, Universitas Gadjah Mada, Jalan Teknika Selatan, Yogyakarta 55281
3Fakultas Pertanian, Universitas Gadjah Mada, Jalan Bulaksumur, Yogyakarta 55281

The term endophyte refers to internal colonization of plant by microorganisms without any pathogenic effects on their hosts. Interaction between endophyte diazotrophic bacteria in sugarcane tissue represents a promising model system for monocot and an endophyte diazotrophic bacteria, as the ability of the diazotroph bacteria to enhance sugarcane growth has been well documented. Diversity of endophyte diazotrophic bacteria was observed and identified by using repetitive-PCR sequence method and sequencing of 16S rDNA. The results showed based on repetitive-PCR sequence methods, that 4 isolates (10.2.2, 10.2.3, 10.3.2.1, and 10.3.2.2) were shown to be very similar to one another. Therefore, isolate 10.2.3 was selected to represent the group for further study. Three isolates (10 K1, 10 K2, and NB12) were found to be different from each other. Phylogenetic analysis, based on 16S rDNA sequences, revealed that isolate 10.2.3, 10 K1, 10 K2, and NB12 were identified to be member of genus Klebsiella, Pseudomonas, Acinetobacter, and Bacillus, respectively.

Key words: endophyte, diazotrophic bacteria, repetitive-PCR, rDNA, sugarcane

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JURNAL MIKROBIOLOGI IPB

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