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SELEKSI DAN IDENTIFIKASI BAKTERI ENDOFIT POTENSIAL PENGHASIL ENZIM PROTEASE DARI TAMAN NASIONAL GUNUNG HALIMUN - (The Selection and Identification of Potential Endophyte Bacteria as Protease Enzyme Producer from Halimun Mount National Park) Melliawati, Ruth; Rohmattusolihat, Rohmattusolihat; Nuryati, Nuryati; Rahmani, Nanik; Yopi, Yopi
Biopropal Industri Vol 7, No 2 (2016)
Publisher : Balai Riset dan Standardisasi Industri Pontianak

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (416.906 KB)

Abstract

Endophytic bacteria have an equal chance to bacteria that live outside the plant tissue as potential bacteria. The selection has done towards 326 bacterial endophyte isolates. This research aimed to find and identify proteolytic potential isolates. The proteolytic selection of endophytic bacteria had done using solid skim milk. The capability of endophytic bacteria to agglomerate milk was tested using liquid skim milk which incubated for 7 days at room temperature. Enzyme production of four selected isolates was made through fermentation in GYS medium. The results showed that 86 isolates have proteolytic potential. Isolate HL.29B.63 had highest protease enzymes activity (65.918 U/mL). Medium optimization was able to increase the enzyme activity into 89.94% (125.04 U/mL). The analysis used 16s rDNA showed that isolate HL.29B.63 was Bacillus amyloliquefacient subs. plantarum strain FZB42.Keywords: endophytic bacteria, fermentation, identification, protease, selection ABSTRAKBakteri endofit mempunyai peluang yang sama dengan bakteri yang hidup diluar jaringan tanaman sebagai bakteri potensial. Seleksi dilakukan terhadap 326 isolat bakteri endofit. Tujuan penelitian ini adalah mencari isolat yang berpotensi proteolitik dan mengidentifikasinya. Seleksi proteolitik terhadap bakteri endofitik menggunakan skim milk padat. Uji kemampuan bakteri endofitik dalam menggumpalkan susu menggunakan medium skim milk cair yang diinkubasi selama 7 hari pada suhu ruang. Produksi enzim terhadap empat isolat terseleksi dilakukan melalui fermentasi dalam medium GYS. Hasilnya menunjukkan bahwa 86 isolat mempunyai potensi proteolitik. Isolat HL.29B.63 mempunyai aktif enzim protease tertinggi (65,918 U/mL). Optimasi medium dapat meningkatkan aktivitas enzim sebesar 89,94% (125,04 U/mL). Analisis menggunakan 16s rDNA menunjukkan bahwa isolat HL.29B.63 adalah Bacillus amyloliquefaciens subs. plantarum strain FZB42.Kata kunci: bakteri endofit, fermentasi, identifikasi, protease, seleksi
Pertumbuhan Bakteri Laut Shewanella indica LBF-1-0076 dalam Naftalena dan Deteksi Gen Naftalena Dioksigenase - (The Growth of Marine Bacteria Shewanella indica LBF-1-0076 in Naphthalene and Naphthalene dioxygenase Gene Detection) Farini, Nuzul; Thontowi, Ahmad; Yetti, Elvi; Suryani, Suryani; Yopi, Yopi
Biopropal Industri Vol 8, No 1 (2017)
Publisher : Balai Riset dan Standardisasi Industri Pontianak

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Abstract

Crude oil exploitation which often occured offshore can cause water pollution in the sea since its contains naphthalene which is a hazardous compounds. This research used marine bacteria LBF-1-0076 that have ability in naphthalene degradation. This research aimed to study the parameter effect of naphthalene and cell concentration toward marine bacteria LBF-1-0076. This research also identified isolate LBF-1-0076 and detected the encode gene of naphthalene dioxygenase. Based on growth test result, the optimum naphthalene degradationby isolate LBF-1-0076 occured in 75 ppm naphthalene concentration with 15cell concentration. The result of 16S rDNA gene analysis showed that LBF-1-0076 was identified as Shewanella indica strain 0102 with identical value 99%. The result of naphthalene dioxygenase gene detection using Polymerase Chain Reaction (PCR) showed that the isolate contained naphthalene dioxygenase gene with size ±377 bp. Therefore, LBF-1-0076 potential as bioremediation agent to solve crude oil contamination in the sea.Keywords:   crude oil, marine bacteria, naphthalene, naphthalene dioxygenase, Shewanella indicaABSTRAKEksploitasi minyak bumi yang sering terjadi di laut mengakibatkan adanya pencemaran minyak di laut. Naftalena merupakan salah satu senyawa dominan berbahaya yang terkandung dalam minyak bumi dan dapat mengakibatkan pencemaran perairan. Penelitian ini menggunakan bakteri laut LBF-1-0076 yang memiliki kemampuan untuk mendegradasi naftalena. Tujuan dari penelitian ini adalah mempelajari pengaruh parameter konsentrasi naftalena dan konsentrasi sel terhadap bakteri laut pendegradasi naftalena LBF-1-0076. Penelitian ini juga bertujuan untuk mengidentifikasi isolat LBF-1-0076 dan mendeteksi gen pengkode naftalena dioksigenase. Berdasarkan hasil uji pertumbuhan, degradasi naftalena yang optimal oleh isolat LBF-1-0076 terjadi pada konsentrasi naftalena 75 ppm dengan konsentrasi sel 15. Hasil analisis gen 16S rDNA menunjukkan isolat LBF-1-0076 teridentifikasi sebagai Shewanella indica strain 0102 dengan nilai keidentikan 99%. Hasil deteksi gen naftalena dioksigenase dengan menggunakan Polymerase Chain Reaction (PCR) menunjukkan bahwa isolat tersebut mempunyai gen naftalena dioksigenase dengan ukuran ±377 bp. Oleh karena itu, isolat LBF-1-076 berpotensi sebagai agen bioremediasi untuk mengatasi masalah pencemaran minyak bumi di laut.Kata kunci: bakteri laut, minyak bumi, naftalena, naftalena dioksigenase, Shewanella indica
Pertumbuhan Optimal Bakteri Laut Pseudomonas aeruginosa LBF-1-0132 dalam Senyawa Piren Safitriani, Safitriani; Thontowi, Ahmad; Yetti, Elvi; Suryani, Suryani; Yopi, Yopi
JURNAL BIOLOGI INDONESIA Vol 13, No 1 (2017): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v13i1.3100

Abstract

ABSTRACTPyrene is a high molecular weight chemical compound belongs to polycyclic aromatic hydrocarbon (PAHs) group that are difficult to degrade by environment. Biodegradation techniques using indigenous marine bacteria are used to be as an effort to reduce pollutants that are carsinogenic. The objectives of this research are to screen of 18 marine bacteria isolates qualitatively by sublimation method and quantitatively by growth test and to optimize degradation activity of marine bacteria isolates by pyrene concentration and cell concentration. Identification by 16S rDNA and phylogenetic tree analysis were conducted to determine the molecular basis of bacterial identity. The result of sublimation showed that 15 isolates were positive result for pyrene degradation and classified to 3 groups. The first group consisted of 5 isolates that can produce clear zone, while the second group are 5 isolates with isolate color changes. The third group have both of activities. Growth test showed that isolate LBF-1-0132 has high potency to degrade pyrene compound. Isolate LBF-1-0132 is capable of degrading pyrene compounds optimally at concentration of 600 ppm and optimum cell concentration of 20. Based on 16S rDNA gene analysis, isolate LBF-1-0132 is Pseudomonas aeruginosa with 98% identity.Keywords :pyrene, marine bacteria, optimization, 16S rDNA identification
Isolats Bakteri Indigenous Penghasil Milk-Clotting Protease untuk Fermentasi Keju Rahmani, Nanik; Sari, Yana Nurita; Palupi, Nurheni Sri; Yopi, Yopi
JURNAL BIOLOGI INDONESIA Vol 9, No 2 (2013): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (316.729 KB) | DOI: 10.14203/jbi.v9i2.166

Abstract

The aims of this research is to isolation of bacteria that potential to produce of milk clotting protease enzymes fromfermented food that will be used as a substitute for rennet in cheese making. There are five food fermentations suchas tauco, tempeh, red oncom, sticky tape, and pickled mustard greens that are used as a source for isolation of bacteriathat could produce milk clotting protease. The results obtained four isolates proteolytic bacteria from two fermentedfood samples, three isolates bacteria from tauco (TCN 1, TCN 2, TCN 3) and one isolate from pickledmustard greens (DSN 1). Based on 16S rDNA, these isolates were identified as Bacillus sp. Bacterial isolate TCN 1has a milk clotting activity of 29.17 U/mL, whereas bacteria isolates of TCN 2, TCN 3 and DSN 1 have activities of70 U/mL achieved at the 24 hours incubation, respectively. The proteolytic activities of bacteria isolates TCN 1,TCN 2, TCN 3 and DSN 1 at the 24 hours fermentation process were 0.0117 U/mL, 0.0021 U/mL, 0.0150 U/mL,and 0.200 U/mL, respectively. The ratio of milk clotting protease activity and the proteolytic activity for bacteriaisolates TCN 1, TCN 2, TCN 3 and DSN respectively were 5402, 175000, 7292, and 3333. This showed that theenzyme from bacterial isolates TCN 2 can be used as an alternative to rennin in cheese making.Keywords: milk clotting protease, cheese, calf rennet, fermentation food
Pemurnian Parsial dan Karakterisasi Amilase dari Bakteri Laut Arthrobacter arilaitensis LBF-003 Rahmasari, Dianti; Wijanarka, Wijanarka; Pujiyanto, Sri; Rahmani, Nanik; Yopi, Yopi
JURNAL BIOLOGI INDONESIA Vol 12, No 1 (2016): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v12i1.2323

Abstract

Starch is an abundant carbon source in nature, and ?-amylase (1, 4-?-D-glucanohydrolase; EC 3.2.1.1), which hydrolyzes ?-1, 4-glucosidic linkage in starch-related molecules. Microbe ?-amylase production is a hydrolytic enzyme and one ofinterest in its microbial production has increased dramatically due to its wide spread use in food, textile, baking anddetergent industries in recent years. Here we report ?-amylase from marine bacterium which was purified andcharacterized, as well as analyzed its hydrolysis product on starch. The enzyme of Arthrobacter arilaitensis partiallypurified by acetone precipitation with 90% and ion exchange chromatography produced specific activity 0.25 U/mg and0.38 U/mg, and it’s purity rate increased until 1.14 fold compared with former crude extract. Purifed extracelluler amilasehad an optimum activity at temperature 50°C and pH 9.0. An apparent molecular mass was between 50-75 kDa, asestimated by zimogram electrophoresis. Hydrolysis products of this enzyme on starch were maltose, maltotriose andmaltoheptaose.Keywords: alfa amylase, marine bacterium, Arthrobacter arilaitensis, purification, charaterization
Karakterisasi Biodegradasi Senyawa Poliaromatik Dibenzothiophene Oleh Bakteri Laut Novosphingobium mathurense LBF-1-0061 Tanjung, Puspasari Noerwan; Yetti, Elvi; Thontowi, Ahmad; Suprihadi, Agung; Purwantisari, Susiana; Yopi, Yopi
JURNAL BIOLOGI INDONESIA Vol 12, No 2 (2016): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1056.931 KB) | DOI: 10.14203/jbi.v12i2.2894

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ABSTRACTDibenzothiophene is one of polycyclic aromatic hydrocarbon (PAH) compound containing sulfur element. This compound has toxicity, mutagenic and quiet persistent in environment. From sreening test, it was known that isolate LBF-1-0061 was potential to degrade dibenzothiophene. The objectives of this study are to study dibenzotiophene degrading capability by marine bacteria isolate LBF-1-0061 using screening test; analysis of dibenzothiophene residue by GC/MS and identifiy the isolate by molecular identification. The result of this research shown that LBF-1-0061 isolate could grow up to 100 ppm of dibenzotiophene. This isolate also presented degrading capability approximately 37.5% of dibenzotiophene in 14 days incubation. Based on partial 16S rRNA gene analysis, LBF-1-0061 was identified 99% as Novosphingobium mathurense strain SM117.Keywords: sea bacteria, biodegradation, dibenzotiofen, hydrocarbon aromatic polisiclic
Keragaman Bakteri Laut Pendegradasi Alkana dan Poliaromatik Hidrokarbon di Pulau Pari Jakarta Thontowi, Ahmad; Yopi, Yopi
JURNAL BIOLOGI INDONESIA Vol 9, No 1 (2013): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (185.283 KB) | DOI: 10.14203/jbi.v9i1.154

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Minyak mentah merupakan salah satu sumber pencemaran di lingkungan laut. Degradasi oleh bakteri memegangperanan penting dalam bioremediasinya. Sejumlah 66 bakteri laut dari Pulau Pari, Kepulauan Seribu Jakarta telahdiisolasi, dianalisa berdasarkan gen 16S rDNA, dan diuji kemampuannya dalam mendegradasi minyak. Berdasarkananalisis gen 16S rDNA diperoleh lima kelompok bakteri pendegradasi minyak, yaitu α-proteobakteria (43.6%), γ-proteobakteria (48.5 %), Flavobakteria (4.5 %), Aktinobakteria (1,5 %), dan Bacillales (1,5%). Bakteribakteritersebut mampu mendegradasi komponen minyak (senyawa alkana dan poliaromatik hidrokarbon). γ-Proteobakteria dan α-proteobakteria mempunyai peran penting dalam bioremediasi minyak di kawasan lingkunganlaut di Pulau Pari. Dari hasil tersebut membuktikan bahwa bakteri pendegradasi minyak dari Pulau Pari sangatberagam.Kata kunci: minyak, laut, bakteri, bioremediasi, alkana, poliaromatik hidrokarbon
Evaluation of Non-Saccharomyces Cerevisiae Strains Isolated from Sea Water Against Inhibitory Compounds for Ethanol Production Thontowi, Ahmad; Putra, Filemon Jalu Nusantara; Yopi, Yopi
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 12, No 2 (2017): August 2017
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v12i2.284

Abstract

An important parameter in industrial bioethanol fermentation is the resistance of yeast to osmotic pressure and inhibitor compounds. Aureobasidium pullulans LBF-3-0074 and Schwanniomyces etchellsii LBF-3-0034 are reported capable to produce ethanol. LBF-3-0034 and LBF-3-0074 are yeast strains isolated from Bali and Lombok sea water. This study aimed to evaluate characteristics of both LBF-3-0034 and LBF-3-0074 strains under the effects of glucose and inhibitor compounds. Both strains were allowed to consume glucose up to 120 mM. Then, these strains were grown with the present of several inhibitors, i.e. 5-hydroxymethyl-2-furaldehyde (5-HMF), furfural, acetic acid, formic acid, and levulinic acid. Results showed that the two yeast strains studied could grow and ferment the sugars under both osmotic and inhibitor stress conditions. As conclusion, Schwanniomyces etchellsii LBF-3-0034 and Aureobasidium pullulans LBF-3-0074 are potential for direct fermentation of lignocellulosic hydrolysate to ethanol.
Production of Manooligomannan from Palm Kernel Cake by Mannanase Produced from Streptomyces Cyaenus Purnawan, Awan; Yopi, Yopi; Irawadi, Tun Tedja
Biosaintifika: Journal of Biology & Biology Education Vol 9, No 1 (2017): April 2017
Publisher : Department of Biology, Faculty of Mathematics and Sciences, Semarang State University . Ro

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15294/biosaintifika.v9i1.9201

Abstract

The increase of public attention to health has prompted researchers to look for new sources of functional food. Palm Cake Kernel (PKC) waste was abundant in Indonesia, Oligosaccharide has an important benefit for human health. Recently oligosaccharide is not only important as an artificial sweetener, but also as a functional food component. This study was aimed to produce oligo-mannan enzymatically from PKC waste using mannanase derived from of Streptomyces cyaenus isolates of indigenous Indonesia. The enzyme concentration was determined by enzyme activity assay while oligo-mannan content in the PKC was analyzed using TLC and HPLC. Mannanase enzyme activity of 1706 U/ml on the second day of agitation 200 rpm at a temperature of 30C Hydrolysis of mannooligomannan by using mannanase produced by streptomyces cyaenus. The optimum mannanase enzyme activity obtained on day 2 with the value of the activity as much of 0.702 U/mL. The protein content of the 2nd day at an agitation speed of 150 rpm, 200 rpm, and 250 rpm, respectively, were 1783, 1950 and 2283 ppm. Streptomyces cyaenus is Indonesian original isolates potentially producing mannanase that can produce mannooligomannan.
Optimization of Substrate and Starter Cell Concentrations for Dibenzothiopene Biodegradation by Indigeneous Marine Bacteria Mauricauda olearia LBF-1-0009, Alcanivorax xenomutants LBF-1-0018, and Stakelama pacifica LBF-1-0031 Yetti, Elvi; Thontowi, Ahmad; Yopi, Yopi
ANNALES BOGORIENSES Vol 21, No 2 (2017): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (646.12 KB) | DOI: 10.14203/ab.v21i2.300

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Dibenzothiophene (DBT) and its derivatives have been widely used as model organic sulfur compounds in petroleum, included their biodegradation process. The abilities of microorganisms to degrade pollutants are significantly influenced by various factors such as microbial species, nutrients and environmental parameters. In this research, we carried out further study to determine optimal condition for DBT biodegradation regarding with substrate and strains cell concentration by several indigenous marine bacteria from Indonesia. These three isolates were belong to Mauricauda olearia, Alcanivorax xenomutants, and Stakelama pacifica, with homology result 99% each. Optimal dibenzothiophene as substrate reached by all isolates is 100 ppm, while cell concentration or microbial numbers that gave highest growth for all isolates is 20 based on conversion of OD600 nm measurement. Â