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Journal : JURNAL BIOLOGI INDONESIA

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)
A NEW INDIGENOUS CYANOMETHANE-DEGRADING BACTERIUM ISOLATED FROM GOLD MINING WASTE WATER Sulistinah, Nunik; Munandar, Hendra; Sunarko, Bambang
JURNAL BIOLOGI INDONESIA Vol 15, No 2 (2019): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v15i2.3807

Abstract

ABSTRACT The gold mining wastewater effluent is potential source for isolating cyanides-degrading bacteria, since cyanide is commonly used in gold extraction process in the mining industry. An indigenous bacterial strain LP3, capable of growing on and utilizing of a high concentration of cyanomethane (up to 1.0 Molar), could be isolated from Cikotok gold mine effluent. Based on 16S rDNA sequence, the strain was identified as Rhodococcus pyridinivorans. During the growth on cyanomethane (CH3CN), ethanamide (CH3ONH2) and ethanoic acid (CH3COOH) were detected in the growth media, indicating that nitrile hydratase and amidase involved in the metabolism of the substrate. The involvement of both enzymes on the conversion of cyanomethane was also proved by our study on cyanomethane biodegradation using whole cells of R. Pyridinivorans LP3. Besides cyanomethane, the R. pyridinivorans LP3 could also utilize various aliphatic, aromatic, heterocyclic nitriles and amides as growth substrates. Base on these results, R. pyridinivorans LP3 is expected to be used as a potential candidate for biological treatment for cyanide-containing wastes, although further research is still needed, before being applied on a field scale.  Keywords: biocatalyst, cyanide degrading bacteria, gold mining, Rhodococcus pyridinivorans LP3
A NEW INDIGENOUS CYANOMETHANE-DEGRADING BACTERIUM ISOLATED FROM GOLD MINING WASTE WATER Sulistinah, Nunik; Munandar, Hendra; Sunarko, Bambang
JURNAL BIOLOGI INDONESIA Vol 15, No 2 (2019): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v15i2.3807

Abstract

ABSTRACT The gold mining wastewater effluent is potential source for isolating cyanides-degrading bacteria, since cyanide is commonly used in gold extraction process in the mining industry. An indigenous bacterial strain LP3, capable of growing on and utilizing of a high concentration of cyanomethane (up to 1.0 Molar), could be isolated from Cikotok gold mine effluent. Based on 16S rDNA sequence, the strain was identified as Rhodococcus pyridinivorans. During the growth on cyanomethane (CH3CN), ethanamide (CH3ONH2) and ethanoic acid (CH3COOH) were detected in the growth media, indicating that nitrile hydratase and amidase involved in the metabolism of the substrate. The involvement of both enzymes on the conversion of cyanomethane was also proved by our study on cyanomethane biodegradation using whole cells of R. Pyridinivorans LP3. Besides cyanomethane, the R. pyridinivorans LP3 could also utilize various aliphatic, aromatic, heterocyclic nitriles and amides as growth substrates. Base on these results, R. pyridinivorans LP3 is expected to be used as a potential candidate for biological treatment for cyanide-containing wastes, although further research is still needed, before being applied on a field scale.  Keywords: biocatalyst, cyanide degrading bacteria, gold mining, Rhodococcus pyridinivorans LP3
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 | 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
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
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 semiâ??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)