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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 PublicationJOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
ISSN1017-7825
Volume25Issue: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.

KeywordNitrilase Hyperthermophilic Bacterium Thermotoga Maritima Temperature Tolerance Aliphatic Dinitriles
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
DOI10.4014/jmb.1502.02032
Indexed BySCI
Language英语
WOS KeywordALCALIGENES SP ECU0401 ; (R)-(-)-MANDELIC ACID ; BIOCHEMICAL-CHARACTERIZATION ; ENANTIOSELECTIVE NITRILASE ; HETEROLOGOUS EXPRESSION ; SUBSTRATE-SPECIFICITY ; EFFICIENT PRODUCTION ; IMINODIACETIC ACID ; PSEUDOMONAS-PUTIDA ; ESCHERICHIA-COLI
WOS Research AreaBiotechnology & Applied Microbiology ; Microbiology
WOS SubjectBiotechnology & Applied Microbiology ; Microbiology
Funding OrganizationOpen 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 IDWOS:000365443800012
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/19897
Collection生化工程国家重点实验室
Affiliation1.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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