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Penguncilan Gen Penyandi Enzim Nitrilase Enam Isolat Bakteri Unggulan Riffiani, Rini; Sulistinah, Nunik; Sunarko, Bambang
JURNAL BIOLOGI INDONESIA Vol 11, No 1 (2015): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (3206.948 KB) | DOI: 10.14203/jbi.v11i1.2155

Abstract

Indonesia sebagai negara tropis memiliki biodiversitas yang sangat tinggi. Keanekaragaman hayati ini diperkirakan mencerminkan keanekaragaman kimiawi sekaligus keragaman genetik yang dapat dimanfaatkan untuk mencari biokatalis baru. Enam isolat bakteri yaitu GLB5, LP3, TPIK, MICC, 23A2, dan 23A2 telah diisolasi dari berbagai limbah industri dan mempunyai potensi sebagai pendegradasi nitril.  Pengucilan, identifikasi dan purifikasi gen penyandi enzim nitrilase dari keenam isolat bakteri tersebut  telah dilakukan. Dari kegiatan penelitian ini 3 isolat bakteri unggulan, yaitu GLB5, LP3, dan TPIK teridentifikasi sebagai Rhodococcus pyridinivorans, sedangkan  MICC teridentifikasi sebagai Bacillus substilis, 23A2 teridentifikasi sebagai Brevibacillus brevis, dan 26A2 teridentifikasi sebagai Microbacterium oxydans. Peta untaian basa nukleutida dari gen penyandi enzim nitrilase dari ketiga isolat yaitu GLB5, LP3, dan TPIK telah terpetakan dengan ukuran gen nitrilase sebesar 960 bp. Hasil analisis dengan BLASTN memperlihatkan bahwa fragmen gen nitrilase yang diamplifikasi dengan primer Nit1101F dan Nit1101R mempunyai homologi yang tinggi terhadap Rhodococcus rhodochrous strain tg1-A6 nitrilase gen dengan persentase kesamaan sebesar 96% . Kata Kunci: Gen, isolasi, nitril, degradasi, enzim 
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 | Full PDF (339.945 KB) | 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
Karakteristik Fisiologis Enzim Nitril Hidratase dan Amidase dalam Sel Corynebacteriurn sp. D5 Sulistinah, Nunik; Kaban, Joseva Sudiati; Sunarko, Bambang
JURNAL BIOLOGI INDONESIA Vol 3, No 4 (2002): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (616.571 KB) | DOI: 10.14203/jbi.v3i4.3323

Abstract

ABSTRACTPhysiological Characteristics of Nitrile-Hydratase and Amidase From Corynebacteriumsp. D5. Nitrile hydratase (NH-ase) of Corynebacterium sp. D5 is inductive enzyme, butamidase is constitutive enzyme.The best inducer for Nitril hydratase is 2% (vlv) acetonitrille.Nitril hydratase and amidase enzymes showed to be capable of degrading low moleculeweight of aliphatic nitriles and amides. The optimum condition of NH-ase ofCorynebacteriurn sp. D5 were found out at pH 6,6 and 30°C while amidase at pH 7,2 & 50 Crespectively. The inhibitor of both enzymes seemed to be ~ gand H~*Key words : Nitrile hydratase, bioconversion, Corynebacterium sp. D5, amidase, acetonitrile,aliphatic nitrile
Karakterisasi Enzim Nitril Hidratase dan Amidase dari Pseudomonas sp. BP3 dalam Biokonversi Adiponitril menjadi Asam Adipat Sunarko, Bambang; Sulistinah, Nunik
JURNAL BIOLOGI INDONESIA Vol 5, No 2 (2008): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (450.693 KB) | DOI: 10.14203/jbi.v5i2.3196

Abstract

ABSTRACTCharacterization of Nitrile Hydratase and Amidase of Pseudomonas sp BP3 in Bioconversionof Adiponitrile to Adipic Acid. Adipic acid is a commercially important compound, primarilyused as precursor for the production of nylon 6.6. It is also used for plasticizer, fibers, and foodadditive. Synthesis of adipic acid by chemical means requires large amount of energy andconcentrated acid. It also produces N2O as by product, which is very toxic and suspectedcauses depletion of the ozone layer. The purpose of this research was to study thebioconversion of adiponitrile by Pseudomonas sp. BP3 and to characterize the involved enzymesin the whole cell. Pseudomonas sp. BP3 was able to utilize adiponitrile as the sole source ofcarbon and nitrogen. It’s doubling time (td) and growth rate constant (?) during the growth inadiponitrile were 2 hours and 0.346/h, respectively. The optimum pH and temperature of nitrilehydratasewere pH 7.0 and 30°C, respectively, while those of amidase were pH 6 and 50°C.Vmax and Ks of nitrile hydratase were 8.3 nM/ml.min. and 55.56 mM, respectively, and ofamidase were 5,9 nM/ml.min and 50 mM. The rate of adiponitrile consumption was 0.245 mM/h and of adipic acid formation was 0.181 mM/h. The yield of bioconversion of adiponitrile andadipamide were about 50 % and 25%, respectively.Key words: Bioconversion, adiponitrile, adipic acid, Pseudomonas sp. BP3, nitrile hydratase,amidase
Pengembangan Sistem Deteksi Senyawa Sianogen dalam Ubi Kayu (Manihot esculenta Crantz) dengan Pendekatan Enzimatis Sulistinah, Nunik; Riffiani, Rini; Sunarko, Bambang
JURNAL BIOLOGI INDONESIA Vol 10, No 1 (2014): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v10i1.332

Abstract

Picrate paper test kit for the semi–quantitative determination of cyanogenic potential was developed in thisexperiment. The method is relatively simple, easy to use and might be applicable in the field by unskilled person.Paper test was attached on tubes containing sample (100 mg) in aquadest (0,5 mL) and then was immediatelycovered tightly and incubated overnight at room temperature. The colour of picrate paper test changed graduallytowards reddish brown, and its colour was compared with standart colour chart which included 0-800 ppm cyanidethat was also developed in this study. The reddish brown colour of paper test was correlated with cyanideconcentration on the sample. In order to obtain a more accurate detection of cyanogenic compound the paper testwas eluted with 5 mL water or aquadest and the absorbance was measured at 510 nm.Keywords: cyanogenic potential, picrate paper test, semi-quantitative method, simple method, cassava (Manihotesculenta Cranz)
Penapisan Mikroba Laut Perombak Senyawa Nitril dan Protein yang Diisolasi Dari Spons di Perairan Ternate Riffiani, Rini; Sulistinah, Nunik
JURNAL BIOLOGI INDONESIA Vol 6, No 3 (2010): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (262.562 KB) | DOI: 10.14203/jbi.v6i3.3143

Abstract

ABSTRACTScreening of Nitrile and Protein-Degrading Marine Bacteria Isolated from Sponge in TernateSea Water. Thirty three marine bacteria have been isolated from marine sponge in Ternate byenrichment culture. Screening bacteria-degrading nitrile was done by microtitter plate methodbased on growth ability tested by Iodonitrotetazolium chloride. Product of nitrile degradationwas determined by Gas Chromatography (GC) and the potential bacteria-degrading proteinwas also screened by using selected media which contained casein. The results showed thattwenty one isolates were able to show the clearing zone in selected media. Five isolatescapable of utilizing acetamide as the sole source of carbon and nitrogen. Acetate and ammoniaproduced for hydrolysis acetonitrile by using resting cell of Lysobacter sp.Key words: Nitrile, bacterium, sponge, Ternate
AKTIVITAS ENZIM 2,4-D MONOOKSIGENASE DARI BERBAGAI MIKROBA [2,4-D monooxygenase Activity of Some Microorganisms] Sulistinah, Nunik
BERITA BIOLOGI Vol 4, No 5 (1999)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (230.887 KB) | DOI: 10.14203/beritabiologi.v4i5.1242

Abstract

Nine cultures both from fungi and bacteria have been selected for testing their 2,4-D monooxygenase activity to degrade 2,4-Dtohlorophenoxyacetic acid (2,4-D). The results showed that all the cultures which were tested grows at 1000 ppm 2,4-D. Three cultures (Trichoderma viride. Asperoillus niqer and Isolat E (unidentified)) of the nine cultures are able to grow at 4000 ppm. T. viride grows well on Minimal Basal Media which contained glucose and 2000 ppm 2,4-D and produced the highest biomass (0.8660 g/l) than the others. The biomass of T. viride grew on MBM (without glucose) and added with 2000 ppm 2,4-D is 0,6520g/l. This indicated that the culture is tolerant to 2,4-D and able to use 2,4-D compound as energy and carbon sources for its growth. But we failed to prove the 2,4-D monooxygenase activity of supernatant of T. viride by measuring the changing of pH-value in the 2,4-D breakdown reaction.
POTENSI Rhodococcus pyridinovorans GLB5 SEBAGAI BIOKATALIS DALAM KONVERSI SENYAWA METHIL SIANIDA DAN PHENIL SIANIDA [Potential of Rhodococcus pyridinovrans GLB5 as Biocatalistin Methyl and Phenyl Cyanide Conversion] Sulistinah, Nunik; Riffiani, Rini; Sunarko, Bambang
BERITA BIOLOGI Vol 15, No 1 (2016)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (3384.218 KB) | DOI: 10.14203/beritabiologi.v15i1.2856

Abstract

Nitrile and amide bioconversion have received increasing attention due to their ability to provide a range of commercially important chemicals. The experiment was conducted to investigate the potential of bacterial isolate GLB5 to convert methyl cyanide and phenyl cyanide. The samples were collected from various industrial waste. Selection of isolates to utilize  these substrates as a sole source of energy, carbon and nitrogen was conducted on 96 whell microtitter plates, based on the growth ability using INT (Iodo nitrotetrazolium chloride) reagent. Based on the growth  pattern, it showed that the bacterial isolate GLB5 grew well and it was capable of utilizing  methyl and phenyl cyanide compound as the sole source of carbon and nitrogen.  The isolate GLB5 was isolated from industrial waste of Batik factory in Cirebon, and  identified as Rhodococcus pyridinovorans. Bioconversion of methyl cyanide using whole cells of R. pyridinovorans GLB5 showed that ethanamide (C2H5NO) and ethanoic acid (C2H4O2) were detected. Formation of ethanamide and ethanoic acid as the product of bioconversion, indicated that the nitrile hydratase and amidase enzymes  involved in the bioconversion process. Phenyl carboxamide (C7H7NO) as the product of phenyl cyanide bioconversion was also detected,  although  in  low  concentration. In this study, R. pyridinovorans GLB5 was capable of completely converting 300 mM methyl cyanide to ±  140 mM ethanoic acid in relatively short times (<60 minutes).
KERAGAMAN BAKTERI PENGHASIL ENZIM PENGHIDROLISIS NITRIL DI PULAU ENGGANO BENGKULU [Diversity of Nitrilase Producing Bacteria in Enggano Island, Bengkulu] Riffiani, Rini; Sulistinah, Nunik
BERITA BIOLOGI Vol 16, No 3 (2017)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (3647.412 KB) | DOI: 10.14203/beritabiologi.v16i3.2243

Abstract

Potential nitrile degrading microbes have been isolated from marine sponge, marine water and soil in Enggano Island. Nitrilase enzyme has a function in degrading nitrile compund. Nitrilases are important industrial enzymes because of their ability to produce biologically active to degrade enantiomers, such as S-(+)-1-(4’-isobutylphenyl) propionic acid (S-(+)-ibuprofen) and R-(-) mandelic acid. Mandelic acids, which are important as pharmaceutical intermediates, can be produced in enantiomerically pure form by the hydrolysis of their corresponding nitrile. The aim of the study was to explore the diversity of nitrile degrading bacteria in Enggano Island, and their ability to utilize nitrile as a substrate growth. Screening of such microbes were carried out by using microtitter plate method based on growth ability tested by INT (Iodonitrotetrazoliumchloride). Degradation product was determined by High Performance Liquid Chromatography (HPLC). Seventy nine bacteria were able to grow on acetamide, acetonitrile, benzonitrile, adiponitrile, mandelonitrile, succinonitrile, lactonitrile, dan benzilcyanide as the sole source of carbon and nitrogen. Two isolates, YIM 56238 and PO69, have shown to enantioselectively hydrolyze racemic mandelonitrile to mandelic acid. Based on 16S rRNA gene identification, these bacteria have the highest sequence similarity to Microccous endophyticus strain YIM 56238 and Rhodococcus pyridinivorans strain PO69.
PERFORMA BAKTERIPADATANAH TERCEMAR PESTISIDA Rahmansyah, Maman; Sulistinah, Nunik
BERITA BIOLOGI Vol 9, No 6 (2009)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (447.45 KB) | DOI: 10.14203/beritabiologi.v9i6.841

Abstract

Preliminary study on bacterial survive in soil containing pesticide has been carried out. Soil samples collected from Lembang and Dieng. The soil deprive from agriculture area that intensively using pesticide, and compared to other samples gathered from forest soil. All samples examined for total bacteria, denitrification bacteria, phosphate solubilizing bacteria, soil induce respiration, urease and phosphatase activities. Pattern of whole parameters in each soil sample configured similarly, but the value performed differently in the same parameters. Total bacterial population in soil samples also inspected after the samples amended with some certain pesticides. Survival bacteria subjected to media amended with insecticide (Propoxur, Diazinon, and Chlorpyrifos), and herbicides (Bromacil and 2,4-D), and correlation of bacterial growth between sample location were varied. Bacterial degrading pesticide particularly isolated from the soil samples containing 1000 ppm Curzate (fungiside) and 500 ppm 2,4-D.The isolates then cultured in the medium containing insecticide and herbicide, and the response on growth observed in 7 days incubation. Bacterial perform were meaningful to reference of soil degrading pesticide residue in agriculture soil; and it would make representative reference in an effort to use bacteria throughout biofertilizer improvement.