• Archives of Pharmacal Research
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• 제11권4호
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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.

• 한국미생물·생명공학회지
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• 제51권1호
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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.

• 한국미생물·생명공학회지
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• 제34권3호
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• pp.204-210
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• 2006

각종 니트릴 화합물은 키랄 의약품의 합성에 사용되는 유용한 중간체이다. 본 연구에서는 토양 분리균 중에서 4-chloro-3-hydroxy butyronitrile(CHBN)기질로부터 고지혈증 치료제인 Atorvastatin을 합성하는 데에 필요한 4-chloro-3-hydroxy butyric acid(CHBAc)를 생성하는 균주 2종류를 선발하였다. 16S rRNA 분석을 통해서 균 동정을 수행한 결과, 모두 Rhodococcus erythropolis에 속하는 것으로 밝혀졌으며, TLC 분석 결과로부터 CHBN 기질을 분해하는 효소는 니트릴 히드라타아제(NHase)와 아미다아제(amidase)인 것으로 추정되었다. 분리균의 CHBN 분해효소는 ${\varepsilon}$-카프로락탐에 의해서 발현이 유도되었으며, 균체와 세포 추출액에서 모두 기질 분해활성을 나타났다. 기존에 보고된 효소의 유전자 염기서열로부터 프리머를 제조하고 PCR을 수행함으로써 분리균으로부터 니트릴 히드라타아제와 아미다아제 유전자를 확보하게 되었다. 발굴된 유전자의 염기서열을 분석한 결과, 이미 보고된 Rhodococcus erythropolis의 니트릴 히드라타아제 ${\alpha}$-서브유니트과 ${\beta}$-서브유니트 및 아미다아제와 96% 이상의 상동성을 보였다. 따라서 CHBN기질은 분리균의 니트릴 히드라타아제와 아미다아제 효소에 의해서 아미(CHBAm)를 거쳐 산(CHBAc)으로 전환되는 것을 알게 되었다.

• 한국의류산업학회지
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• 제18권6호
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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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• 제24권7호
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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.