Expression and Characterization of a Novel Nitrilase from Hyperthermophilic Bacterium Thermotoga maritima MSB8 | |
Chen, Zhi1; Chen, Huayou1,2; Ni, Zhong1; Tian, Rui1; Zhang, Tianxi1; Jia, Jinru1; Yang, Shengli1 | |
2015-10-01 | |
Source Publication | JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
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ISSN | 1017-7825 |
Volume | 25Issue:10Pages:1660-1669 |
Abstract | The present study describes the gene cloning, overexpression and characterization of a novel nitrilase from hyperthermophilic bacterium Thermotoga maritima MSB8. The nitrilase gene consisted of 804 base pairs, encoding a protein of 268 amino acid residues with a molecular mass of 30.07 kDa after SDS-PAGE analysis. The optimal temperature and pH of the purified enzyme were 45 degrees C and 7.5, respectively. The enzyme demonstrated good temperature tolerance, with 40% residual activity after 60 min of heat treatment at 75 degrees C. The kinetic constants V-max and K-m of this nitrilase toward 3-cyanopyridine were 3.12 mu mol/min/mg and 7.63 mM, respectively. Furthermore, this novel nitrilase exhibited a broad spectrum toward the hydrolysis of the aliphatic nitriles among the tested substrates, and particularly was specific to aliphatic dinitriles like succinonitrile, which was distinguished from most nitrilases ever reported. The catalytic efficiency k(cat)/K-m was 0.44 /mM/s toward succinonitrile. This distinct characteristic might enable this nitrilase to be a potential candidate for industrial applications for biosynthesis of carboxylic acid. |
Keyword | Nitrilase Hyperthermophilic Bacterium Thermotoga Maritima Temperature Tolerance Aliphatic Dinitriles |
Subtype | Article |
WOS Headings | Science & Technology ; Life Sciences & Biomedicine |
DOI | 10.4014/jmb.1502.02032 |
Indexed By | SCI |
Language | 英语 |
WOS Keyword | ALCALIGENES SP ECU0401 ; (R)-(-)-MANDELIC ACID ; BIOCHEMICAL-CHARACTERIZATION ; ENANTIOSELECTIVE NITRILASE ; HETEROLOGOUS EXPRESSION ; SUBSTRATE-SPECIFICITY ; EFFICIENT PRODUCTION ; IMINODIACETIC ACID ; PSEUDOMONAS-PUTIDA ; ESCHERICHIA-COLI |
WOS Research Area | Biotechnology & Applied Microbiology ; Microbiology |
WOS Subject | Biotechnology & Applied Microbiology ; Microbiology |
Funding Organization | Open Funding Project of National Key Laboratory of Biochemical Engineering ; National Key Basic Research Program of China (973 Program)(2011CBA00800) ; Key Agriculture Support Project of Jiangsu Province, China(BE2013400) |
WOS ID | WOS:000365443800012 |
Citation statistics | |
Document Type | 期刊论文 |
Identifier | http://ir.ipe.ac.cn/handle/122111/19897 |
Collection | 生化工程国家重点实验室 |
Affiliation | 1.Jiangsu Univ, Inst Life Sci, Zhenjiang 212000, Peoples R China 2.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China |
Recommended Citation GB/T 7714 | Chen, Zhi,Chen, Huayou,Ni, Zhong,et al. Expression and Characterization of a Novel Nitrilase from Hyperthermophilic Bacterium Thermotoga maritima MSB8[J]. JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY,2015,25(10):1660-1669. |
APA | Chen, Zhi.,Chen, Huayou.,Ni, Zhong.,Tian, Rui.,Zhang, Tianxi.,...&Yang, Shengli.(2015).Expression and Characterization of a Novel Nitrilase from Hyperthermophilic Bacterium Thermotoga maritima MSB8.JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY,25(10),1660-1669. |
MLA | Chen, Zhi,et al."Expression and Characterization of a Novel Nitrilase from Hyperthermophilic Bacterium Thermotoga maritima MSB8".JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY 25.10(2015):1660-1669. |
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Expression and Chara(2908KB) | 期刊论文 | 出版稿 | 限制开放 | CC BY-NC-SA | Application Full Text |
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