• Archives of Pharmacal Research
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• v.11 no.4
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• pp.285-291
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• 1988

2-mercaptoethanol, thioglycolic acid, glutathione, 3-mercapto-1, 2-propanediol and 3-mercapto-2-butanol showed catalytic activities on the hydrolysis of $\alpha$-amino-phenylacetonitrile to phenylglycineamide at the rate of 12.19 $\times$ $10^{-2}$, 8.03 $\times$ $10^[-2}$, 6.83 $\times$ $10^{-2}$, 8.60 $\times$ $10^{-2}$ and 6.04 $\times$ $10^{-2}$ mM $min^{-1}$, respectively. hte hydrolysis rate was faster in buffer than in water. The hydrolysis of the nitrile compound to phenylglycine was limited.

• Microbiology and Biotechnology Letters
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• v.51 no.1
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• pp.69-82
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• 2023

Nitrilases are a hydrolase group of enzymes that catalyzes nitrile compounds and produce industrially important organic acids. The current objective is to optimize nitrilase production using statistical methods assisted with artificial intelligence (AI) tool from novel nitrile degrading isolate. A nitrile hydrolyzing bacteria Bacillus subtilis AGAB-2 (GenBank Ascension number- MW857547) was isolated from industrial effluent waste through an enrichment culture technique. The culture conditions were optimized by creating an orthogonal design with 7 variables to investigate the effect of the significant factors on nitrilase activity. On the basis of obtained data, an AI-driven support vector machine was used for the fitted regression, which yielded new sets of predicted responses with zero mean error and reduced root mean square error. The results of the above global optimization were regarded as the theoretical optimal function conditions. Nitrilase activity of 9832 ± 15.3 U/ml was obtained under optimized conditions, which is a 5.3-fold increase in compared to unoptimized (1822 ± 18.42 U/ml). The statistical optimization method involving Plackett Burman Design and Response surface methodology in combination with an AI tool created a better response prediction model with a significant improvement in enzyme production.

• Microbiology and Biotechnology Letters
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• v.34 no.3
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• pp.204-210
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• 2006

Ethyl (S)-4-chloro-3-hydroxybutyrate is a useful intermediate for the synthesis of Atorvastatin, a chiral drug to hypercholesterolemia. In this research, two 4-chloro-3-hydroxybutyro-nitrile-degrading strains were isolated from soil sample. They were identified as Rhodococcus erythropolis strains by 16S rRNA analysis. The nitrile-degrading enzyme(s) were suggested to be nitrile hydratase and amidase rather than nitrilase from the result of thin layer chromatography analysis. The corresponding genes were obtained by PCR cloning method. The predicted protein sequences had identities more than 96% with nitrile hydratase ${\alpha}-subunit$, nitrile hydratase ${\beta}-subunit$, and amidase of R. erythropolis. The 4-chloro-3-hydroxybutyronitrile-hydrolyzing activities in both strains were increased dramatically by ${\varepsilon}-caprolactam$ which was known as good inducer for nitrile hydratase. Both intact cells and cell-free extract could hydrolyze the nitrile compound. So, the intact cell and the enzymes could be used as potential biocatalyst for the production of 4-chloro-3-hydroxybutyric acid.

• Fashion & Textile Research Journal
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• v.18 no.6
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• pp.889-896
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• 2016

This study aims to evaluate the hydrolytic activity of a commercial nitrilase and optimize nitrilase treatment conditions to apply eco-friendly finishing on acrylic fabrics. To assess the possibility of hydrolyzing nitrile bonds in acrylic fabric using a commercial nitrilase, the amounts of hydrolysis products, ammonia and carboxylate ions, were measured. The treatment conditions were optimized via the amount of ammonia. The formation of carboxylate ions on the fabric surface was detected by X-ray photoelectron spectroscopy and wettability measurements. After nitrilase treatment, ammonia was detected in the treatment liquid; thus, nitrilase hydrolyzed the nitrile bonds in acrylic woven fabric. The largest amount of ammonia was released into the treatment liquid under the following conditions: pH 8.0, $40^{\circ}C$, and a treatment time of 5 h. The formation of carboxylate ions on the acrylic woven fabric surface by nitrilase hydrolysis was proven by the increased O1s content measuring of XPS analysis. From comparison of the results of nitrilase and alkaline hydrolysis, the white index and strength of the alkali-hydrolyzed acrylic fabric decreased, whereas those of the nitrilase-hydrolyzed samples were maintained. The nitrilase hydrolysis improved the sensitivity of acrylic fabrics to basic dye similarly to alkaline hydrolysis without the drawbacks of yellowing and decreased strength caused by alkaline hydrolysis.

• Journal of Microbiology and Biotechnology
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• v.24 no.7
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• pp.936-942
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• 2014

Using racemic (R,S)-2-(4-chlorophenyl)-3-methylbutyramide, an intermediate for the chiral pyrethroid insecticide Esfenvalerate, as a sole nitrogen source in a minimal medium, several strains with high enatioselectivity (${\geq}98%$) were isolated by enrichment techniques. One of the strains, LG 31-3, was identified as Burkholderia multivorans, based on physiological and morphological tests by a standardized Biolog station for carbon source utilization. A novel amidase was purified from B. mutivorans LG 31-3 and characterized. The enzyme exhibited (S)-selective amidase activity on racemic (R,S)-2-(4-chlorophenyl)-3-methylbutyramide. Addition of the racemic amide induced the production of the enantioselective amidase. The molecular mass of the amidase on SDS-PAGE analysis was shown to be 50 kDa. The purified amidase was subjected to proteolytic digestion with a modified trypsin. The N-terminal and internal amino acid sequences of the purified amidase showed a high sequence homology with those deduced from a gene named YP_366732.1 encoding indole acetimide hydrolase from Burkholderia sp. 383.