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Medium Chain and Long Chain Alkanes Hydroxylase Producing Whole Cell Biocatalyst From Marine Bacteria Thontowi, Ahmad; Yetti, Elvi; Yopi, Yopi
ANNALES BOGORIENSES Vol 22, No 1 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ab.v22i1.329

Abstract

Alkanes are  major component of crude oil that could be hydrolyzed by the enzyme of alkane hydroxylase. The are three types of alkane hydroxylase based on the chain length of alkane such as short-chain length/SCL (C2-C4), medium-chain length/MCL (C5-C17), and long-chain length/LCL (C>18). The aims of this study were to characterize and identify alkanes-degrading bacteria from these bacteria. The 30 strains from marine were grown on MCL (Pentane-C5H12, Decane-C10H22, and Pentadecane-C15H32) and LCL (n-Paraffin-C12H19C17 and branch of Pristane-C19H40). The study showed twenty-nine isolates have the ability to degrade alkanes compounds, whereas 14 isolates have grown ability on MCL and LCL medium, 11 isolates have the ability to grow on MCL and n-LCL, 3 isolates have the ability only to grow on MCL medium and 1 isolate has the ability only grow on n-LCL medium. The growth test result indicated that 29 isolates have medium-chain alkane monooxygenase and long-chain alkane hydroxylase. Based on 16S rDNA gene analysis, we obtained twenty nine of oil- degrading bacteria, namely a-proteobacteria (57 %), g-proteobacteria (30 %), Flavobacteria (7 %), Bacilli (3%) and Propionibacteriales (3 %). g-Proteobacteria and a-proteobacteria which seems to play an important role in the alkane biodegradation.
Penapisan dan Optimasi Pertumbuhan Bakteri Laut yang Berpotensi sebagai Hidrokarbonoklastik PAH Fenotiazin Yetti, Elvi; Thontowi, Ahmad; Yopi, Yopi
Jurnal Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 11, No 2 (2016): Desember 2016
Publisher : Balai Besar Riset Pengolahan Produk dan Bioteknologi Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/jpbkp.v11i2.297

Abstract

Fenotiazin merupakan senyawa dari kelompok hidrokarbon aromatik polisiklik atau polycyclic aromatic hydrocarbon (PAH) yang terkandung di dalam minyak mentah. Fenotiazin bersifat persisten dan mudah terbakar di lingkungan. Selain itu fenotiazin juga menyebabkan iritasi kulit, hepatitis, dan anemia terhadap manusia. Bakteri laut memiliki kemampuan untuk mendegradasi senyawa PAH. Tujuan penelitian ini adalah menyelek si bakteri laut yang berpotensi sebagai hidrokarbonoklastik fenotiazin dan melakukan optimasi konsentrasi fenotiazin untuk studi biodegradasinya. Seleksi isolat dilakukan pada media padat dengan metoda sublimasi dan media cair dengan uji pertumbuhan. Hasil seleksi awal dengan metoda sublimasi menunjukkan 32 isolat sebagai kandidat bakteri pendegradasi fenotiazin. Isolat-isolat ini dikelompokkan menjadi 3 kelompok berdasarkan indikatornya dalam seleksi dengan media padat yaitu isolat yang dapat mengubah warna media, membentuk zona bening, dan isolat yang memiliki kemampuan keduanya. Seleksi dengan menggunakan uji pertumbuhan menunjukkan bahwa isolat LBF-1-0057 yang teridentifikasi sebagai Pseudomonas aeruginosa strain MCCB102 dan isolat LBF-1-0126 memiliki pertumbuhan terbaik dari kelompok isolat yang mengubah warna media. Isolat LBF-1-0102 yang teridentifikasi sebagai Pseudomonas balerica BerOc6 dan isolat LBF-1-0133 yang teridentifikasi sebagai P. aeruginosa ATCC10145 merupakan isolat terbaik dari kelompok zona bening, sedangkan isolat LBF-1-0115 adalah isolat dengan pertumbuhan tertinggi di kelompok berindikator keduanya. Konsentrasi optimum fenotiazin untuk P. aeruginosa strain MCCB102, P. aeruginosa ATCC10145, dan isolat LBF-1-0115 adalah 500 ppm; sedangkan 250 ppm merupakan konsentrasi optimum untuk isolat LBF-1-0126 dan P. balerica BerOc6.
PURIFICATION AND PROPERTIES OF MANNANASE FROM Aspergillus ustus BL5 Thontowi, Ahmad; Rahmani, Nanik; Andriani, Ade; Yopi, -
Teknologi Indonesia Vol 37, No 1 (2014)
Publisher : LIPI Press

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (16.051 KB) | DOI: 10.14203/jti.v37i1.214

Abstract

Strain of BL5 was reported as a mannanase producer. The purposes of this study are identification, purification and characterization of mannanase from BL5. The Internal Transcribed Spacer (ITS) regions analysis showed that BL5 strain have 93% similarity with Aspergillus ustus isolate UOA/HCPF 9236. An extracellular mannanase from the culture supernatant of a fungus A. ustus BL5 was purified. SDS-PAGE of the purified enzyme showed a single protein band of molecular mass 50 to 51 kDa. The mannanase exhibited optimum catalytic activity at pH 7.0 and 55C. The metal ions Ca2+, Cu2+ and SDS inhibited complete enzyme activity. The metal ion Mg2+ and EDTA increased complete enzyme activity. The value of Vmax = 5,88 ?mol mannose/min/ml and Kmax = 0.64 mg/ml. Mannanase of A. ustus BL5 be able to hydrolyzed porang mannan.
OPTIMIZATION PRODUCTION OF XYLANASE BY Bacillus pumilus IN EMPTY FRUIT BUNCH USING 3 L BIOREACTOR Wijaya, Hans; Thontowi, Ahmad; Yopi, Yopi
Teknologi Indonesia Vol 40, No 1 (2017)
Publisher : LIPI Press

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Indigenous biomasses, such as empty fruit bunch, contain high xylan and are useful to produce raw xylanase, where this biomasses are very abundant in Indonesia. The focus of this research is to scale up the fermentation product into 3 litter feeds and to record the highest enzyme activity. Fermentation process has been done by using stirred fermentor MBI Winpact One Fermentation System. The isolate marine bacterium Bacillus pumilus has been used to degrade empty fruit bunch to produce raw xylanase. The result for different agitation speed showed that at 150 rpm has the highest xylanase activity. The effect of different aeration flows gives at a rate of 4 VVM achieved the highest enzyme activity and equal to 7.61 U/mL with cell growth equal to 2.06. The Sigma Antifoam C has been successfully being used as defoamer in the production fermentor and can be tolerated by Bacillus pumilus. The comparisons of different medium to generate different enzyme by Bacillus pumilus has achieved xylanase, mannanase, and cellulase, where xylanase has the highest enzyme activity among all of the enzymes produced.
Efek Sumber Karbon Berbeda terhadap Produksi â-Glukan oleh Saccharomyces Cerevisiae pada Fermentor Air Lift Kusmiati, Kusmiati; Thontowi, Ahmad; Nuswantara, Sukma
Jurnal Natur Indonesia Vol 13, No 2 (2011)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (94.458 KB) | DOI: 10.31258/jnat.13.2.138-145

Abstract

The need of â-glucan is increasing in food, medicine and cosmetic industry, because it becomes anticancer,antitumor and antiaging, increases immunosystem, and decreases cholesterol content in blood. The cell walls ofS. cerevisiae contain 80-90% polysaccharides that posses â-glucan. This research was aimed to obtain appropriatecarbon sources to increase the production of â-glucan. The carbon sources used were glucose, glucose commercial,sucrose and molases. The fermentation process was done by using air lift fermentor. The steps of fermentatonincluded regeneration of S. cerevisiae strain, preculture, fermentor preparation and running fermentor for 84hours. Sampling of S. cerevisiae culture was determined the cell growth by optical density (OD) usingspectrophotometer UV/VIS at ë 550 nm. The protein content was determined by Lowry method at ë 755 nm and thetotal glucose was measured by phenol sulphate method at ë 490 nm. The measurement result of cell growthshowed that the high intensity of S. cerevisiae in medium contain molases, but it did not show significant effectwhen compare to other carbon sources. The protein and carbohydrate contain in medium tended to decrease. Theresult of â-glucan on glucose, sucrose, glucose commercial and molases were 933,3, 1100, 1000, and966,7 mg/l. It can be concluded that sucrose and glucose commercial can replace the glucose to produce of â-glucan, because they are cheaper and easier to get. Beside that, molases can be used as an alternative carbonsource because it can produce of â-glucan as well as glucose.
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
PENCIRIAN PRODUKSI AMILASE OLEH SACCAROMYCES CEREVISIAE W303A REKOMBINAN Thontowi, Ahmad; Puspaningsih, Ni Nyoman Tri; Hadi, Sofjan; Purkan, Purkan; Ni'mahtuzahroh, Ni'mahtuzahroh; Irawan, Bambang
JURNAL BIOLOGI INDONESIA Vol 3, No 3 (2002): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v3i3.3464

Abstract

ABSTRACTCharacterization of Amylase Production by Saccharomyces cerevisiae W303A Recombinants. Cloning of amylase gene from Endomycopsis fibuligera ITB.R.cc.64 into S. cerevisiae W303a can effectively increase the yeast function to digest starch directly into ethanol. Production of amylase by S. cerevisiae W303a recombinants (I and P) were done by growing in yeast peptone starch (YPS) medium. The result showed that the recombinants could be produced of amylase by gave clear zone after staining by iodium vapor. The optimum condition of production of amylase by S. cerevisiae W303a recombinants were pH 7.0, 40?C temperature incubation, and gave maximum activity after 36 hours incubation. Amylase activity of I was higher than P recombinant for these condition respectively.Key words: Characterization, amylase, S. cerevisiae W303a
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 | DOI: 10.14203/jbi.v12i2.2894

Abstract

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
METABOLISME BENZONITRIL OLEH FLAVOBACTERIUM SP. NUB 1 Sulistinah, Nunik; Sunarko, Bambang; Thontowi, Ahmad
JURNAL BIOLOGI INDONESIA Vol 3, No 3 (2002): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v3i3.3472

Abstract

ABSTRACTMetabolism of Benzonitriles by Flavobacterium sp. NUB 1. Flavobacterium sp. NUB 1 was isolated from industrial waste of PT. Petrokimia Gresik. The bacterium was able to utilize benzonitrile and acetonitrile and propionitril as the sole source of carbon and nitrogen. Growth on benzonitrile gave higher growth rate and biomass yield than growth on acetonitrile and propionitrile. When Flavovobacterium sp. NUB1 grew on benzonitril 15 mM , the doubling time is 9 hours 54 minutes and the specific growth rate (?) was 0,07 h-1. Whole cell of Flavobacterium sp. NUB 1 could hydrolyzed aromatic and aliphatic nitriles. The bacteria isolate has ability in metabolism of acetonitrile greater than benzonitrile. Activity of nitrile hydratase and amidase are more dominant than nitrilase in metabolism of benzonitrile.Key words: Biodegradation, benzonitril, Flavobacterium sp. NUB 1, nitrile-hydratase,amidase, nitrilase
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 | DOI: 10.14203/jbi.v9i1.154

Abstract

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