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Polyaromatic Hydrocarbon Degradation and Dioxygenase Gene Detection from Alteromonas alvinellae Bt05 Thontowi, Ahmad; Rahmani, Nanik; Yopi, Yopi
ANNALES BOGORIENSES Vol 17, No 1 (2013): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/75

Abstract

Bt05 is marine bacterium which was isolated from the Jakarta Bay, Indonesia. The aim of this study was to characterize PAHs-degrading property, molecular identification by partial analysis of 16S rRNA gene and to partially analyze dioxygenase gene of Bt05 isolate. Our further study on this isolate revealed that it could degrade three PAHs (phenanthrene, dibenzothiophene, fluorene) between 60%–90% within 11 days at 100 ppm level. This finding indicated the potential of the isolate for bioremediation of PAHs. The isolate was identified as Alteromonas alvinellae by phylogenetic analysis of 16S rRNA gene sequence. Sequence analysis of the PCR product of PAH dioxygenase genes amplified using two primer set (iiDA and ppAH) of the isolate were identified 97% as naphthalene dioxygenase gene (phaAc) and 58% as 1,2-dioxygenase.
Isolation and Screening of Surfactant-producing Bacteria from Indonesian Marine Environments and Its Application on Bioremediation Susilaningsih, Dwi; Okazaki, Fumiyoshi; Yopi, Yopi; Widyastuti, Yantyati; Harayama, Shigeaki
ANNALES BOGORIENSES Vol 17, No 2 (2013): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (500.383 KB) | DOI: 10.1234/80

Abstract

Isolation and screening have been undertaken on oil-degrading microbes from Indonesian marine environments. During screening process it has been found many bacterial isolates capable of degrading crude oil. Hence, study has been focused on the biodiversity of biosurfactant-producing bacterial species in Indonesian marine environment and its function for remedial the pollutant in marine and soil areas. A total of 103 out of 463 isolates showed positive surfactant-degrading properties. By means of partial 16S rRNA gene analyses, it has been found that the majority of taxa are related to Alcanivorax, Pseudomonas, Bacillus, Bortetela, Brucella, Acenitobacter, Staphia, Lysobacter, and Talasosophira. Biosurfactant properties assay showed that they were capable of lowering the surface- and interfacial water tension from 74 mN/m to 40-65 mN/m and from 24 mN/m to 6-10 mN/m, respectively. In addition, most of the surfactants were capable of emulsifying hydrocarbon (crude oil) of 0.01 to 0.15 units, comparable to 0.08 units of synthetic surfactant (20% Tween). Further observation showed that the majority of the surfactants were able to degrade a long chain of alkane, but not branched alkane, with a recovering rate of 20-80%. The application of the surfactant towards oil polluted model beach was done in laboratory scale and showing the surfactant obtained from microbial broth cultures capable for recovering the oil pollutant significantly, compared to the control (without addition microbial broth).  
Alkane Degradation and Detection of Mono-xygenase Gene from Alcanivorax sp. from Jakarta Bay Thontowi, Ahmad; Yopi, Yopi
ANNALES BOGORIENSES Vol 15, No 2 (2011): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/48

Abstract

Alkanes is a major component of crude oil that can be hydrolized by enzyme alkane monooxygenase from bacteria. Nine oil-degrading bacteria were analyzed their capability to degrade alkanes (pristane and paraffin). The result of growth test on paraffin and pristane were showed that 9 isolates could be devided into two groups. First group (BL09, BL31 and BL45) could degrade both paraffin and pristane, and second group (BL01, BL06, BL44, BL057, BL058 and BL071) preferred to degrade paraffin than pristane. Three isolates (BL09, BL31 and BL45) have activity to decrease paraffin and pristane until less 50% remain. Based on homology analysis of 16S rRNA gene sequences showed that isolates No. BL09, BL31 and BL45 were identified as Alcanivorax sp. and the partial sequences of the alkB gene from those three isolates are showing 66-68% of identity compare with some mono-oxygenase gen from database of genbank.Keywords: biodegradation, alkane, monooxygenase, cloning, alkB
Medium Optimization for Antimicrobial Production By Newly Screened Lactic Acid Bacteria Rohmatussolihat, Rohmatussolihat; Lisdiyanti, Puspita; Yopi, Yopi; Widyastuti, Yantyati; Sukara, Endang
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.v22i1.322

Abstract

Lactic acid bacteria (LAB) are important for prevention of spoilage and pathogenic bacterial growth in foods due to their ability to generate antimicrobial substances. The objective of this study was to screen LAB for antimicrobial activity and to optimize culture medium for antimicrobial production using Response Surface Methodology (RSM) with Central Composite Design (CCD). Optimization of antimicrobial production of selected LAB was conducted with different combinations of glucose, NaCl, inoculum, and temperature. Our experimental results showed that from 129 LAB isolates, 55 showed significant inhibition against Bacillus subtilis, Escherichia coli, Micrococcus luteus, Staphylococcus aureus, Aspergillus niger, and Candida albicans. No isolates inhibited the growth of Aspergillus flavus. Lactobacillus plantarum LIPI13-2-LAB011 was selected for further study on culture medium optimization to inhibit the growth of C. albicans. From statistical analysis, the production of antimicrobial substances was significantly influenced by temperature, NaCl, and concentration of glucose. Furthermore, the optimum concentrations of glucose, concentration of inoculum, temperature, and NaCl were 1.63 %, 3.03%, 33.74°C, and 3.4%, respectively, with a maximum predicted inhibition index of 1.916, which increased 3.56-fold compared to that obtained in medium before optimization processes. The result was confirmed as when the optimum concentration of nutritions used, the inhibition index increased 3.12-fold.
Glucoamylase Production by Aspergillus awamori KT-11 In Solid State Fermentation Using Cassava Peel as Substrate Perwitasari, Urip; Nuryati, Nuryati; Melliawati, Ruth; Yopi, Yopi
ANNALES BOGORIENSES Vol 21, No 1 (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 (302.187 KB) | DOI: 10.14203/ab.v21i1.290

Abstract

In order to utilization of cassava peel waste this study tries to produce glucoamylase by solid state fermentation with Aspergillus awamori KT-11. Composition medium and drying technique are affecting the glucoamylase production. The highest glucoamylase activities were from cassava peel plus mineral medium. Activity glucomaylase in cassava peel plus mineral medium by oven drying was 365 U/mL and freeze dring was 452 U/mL.  It is conclud cassava peel plus mineral is a better substrate for glucoamylase production from A. awamori KT-11 in solid state fermentation. Powder of glucoamylase also proved capable of hydrolyzing starch-based biomass. 
Optimization of Culture Conditions for Production of β-Mannanase by Strain Nonomuraea sp. ID06-379 using Submerged Substrate Fermentation Ratnakomala, Shanti; Yopi, Yopi; Suhartono, Maggy T; Meryandini, Anja; Prasetya, Bambang
ANNALES BOGORIENSES Vol 18, No 2 (2014): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (685.031 KB) | DOI: 10.1234/96

Abstract

The objective of this study was to investigate the effect of media compositions on the production of β-mannanase by Nonomuraea sp.ID06-379. The study was focused on the influence of carbon, nitrogen,phosphorus and detergents on β-mannanase synthesis through manipulating media compositions on production medium. The results indicated that for carbon sources, locus bean gum (0.745 ± 0.036 U/ml) showed maximum mannanase activity. Malt extract was the best nitrogen source for producing β-mannanase (1.075 ± 0.006 U/ml),(NH4)2HPO4 as phosphate source (1.733 ± 0.026 U/ml) and Tween 80 (1.145 ± 0.003 U/ml) as surfactants effect on increasing permeability of bacterial cell membrane, enhancing membrane transport and excretion of extracellular enzymes into the production media. The results showed that 1% malt extract, 0.5% locus bean gum and 0.05% (NH4)2HPO4 were good substances for nitrogen source, carbon source and phosphate respectively. The highest production of β-mannanase by Nonomuraea sp. ID06-379 (5.33 U/mg) was reached in the medium optimization (Vogel’s minimal medium) contained the following ingredients: 0.5% locus bean gum, 1% malt extract and 0.05% (NH4)2HPO4, under submerged fermentation with shaking at 120 rpm and 28 C for 2 days incubation.
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.
The Investigation of Polycyclic Aromatic Hydrocarbon and Oil Degrading Bacteria Isolated from The Marina Port Ancol, Jakarta Bay Lisdiyanti, Puspita; Yopi, Yopi; Murniasih, Tutik
ANNALES BOGORIENSES Vol 15, No 2 (2011): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/45

Abstract

Polycyclic Aromatic Hydrocarbons (PAHs) as well as crude oil are widespread environmental pollutants. The contamination of air, soil, freshwater (surface water and groundwater), and marine environments by PAHs as well as crude oil has been reported. Of concern to public health is the fact that many PAHs or their metabolites are mutagenic, carcinogenic, or both. North Java coastal area such as Jakarta Bay is the polluted marine area in Indonesia as a result from anthropogenic wastes and the oil spill. Although evaporation and photo-oxidation play an important role in oil detoxification, ultimate and complete degradation is accomplished mainly by marine micro flora, and being dominant in this function. Certain bacteria are well-known could consume and degrade the PAHs as well as crude oil. Therefore investigating the potential PAH and oil degrading marine bacteria is important. In this study, we collected sample from oil polluted area in Marina Port Ancol, Jakarta Bay and isolated four PAH substrates and Arabian crude oil degrading marine bacteria using enrichment method and direct isolation method. As result, 223 strains could degrade PAHs, among these strains, 94 strains could degrade phenanthrene, 23 strains degrade fluoranthene, 92 strains could degrade dibenzotiophen, 14 strains could degrade phenotiazin and 106 isolates degrade crude oil.Key words: polycyclic aromatic hydrocarbons, crude oil, degrading bacteria, bioremediation.
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.
ENZYMATIC HYDROLYSIS OF THE FEC 25 AND ROTI CASSAVA STARCH (Manihot esculenta) VARIETIES BY α-AMYLASE FROM A MARINE BACTERIUM (BREVIBACTERIUM SP.) rahmani, nanik; Andriani, Ade; Hartati, Sri; Yopi, Yopi
Teknologi Indonesia Vol 39, No 3 (2016)
Publisher : LIPI Press

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jti.v39i3.295

Abstract

Cassava (Manihot esculenta) is one of the food sources that is familiar for Indonesian society. Carbohydrates of cassava can be enzymatically hydrolyzed into small oligosaccharides that can be used as a material for functional food components production. Starch from the FEC 25 and roti cassava starch have been hydrolyzed by α-amylase from marine bacterium, Brevibacterium sp. for maltooligosaccharides production. The best hydrolysis reaction condition of the FEC 25 cassava starch were starch concentration of 6.0% (w/v), the ratio of α-amylase and starch 1:1, 50 mM of sodium phosphate buffer pH 6.6, the reaction temperature at room temperature (30oC) and the reaction time of 8 hours with the highest reducing sugar value of 21.675 ppm. While the best hydrolysis of the Roti cassava starch were starch concentration of 6% (w/v), the ratio of α-amylase and starch 1:1, 50 mM of sodium phosphate buffer pH 6.6 and the reaction temperature at 50oC, the reaction time of 8 hours with the highest reducing sugar value of 13.278 ppm. The results of maltooligosaccharides analysis using thin layer chromatography (TLC) showed that the type of maltooligosaccharides formed on hydrolysis the FEC 25 cassava starch are glucose, maltose and maltotriosa, while Roti cassava starch are glucose, maltose, maltotriose, and maltopentaose. The formation of maltooligosaccharides showed that both of cassava starch can be hydrolyzed by α-amylase from marine bacterium Brevibacterium sp.