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Clostridium thermocellum Nitrilase Expression and Surface Display on Bacillus subtilis Spores
Chen, Huayou1,2; Zhang, Tianxi1; Sun, Tengyun1; Ni, Zhong1; Le, Yilin1; Tian, Rui1; Chen, Zhi1; Zhang, Chunxia1
2015
Source PublicationJOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY
ISSN1464-1801
Volume25Issue:6Pages:381-387
Abstract

Nitrilases are an important class of industrial enzymes. They require mild reaction conditions and are highly efficient and environmentally friendly, so they are used to catalyze the synthesis of carboxylic acid from nitrile, a process considered superior to conventional chemical syntheses. Nitrilases should be immobilized to overcome difficulties in recovery after the reaction and to stabilize the free enzyme. The nitrilase from Clostridium thermocellum was expressed, identified and displayed on the surface of Bacillus subtilis spores by using the spore coat protein G of B. subtilis as an anchoring motif. In a free state, the recombinant nitrilase catalyzed the conversion of 3-cyanopyridine to niacin and displayed maximum catalytic activity (8.22 units/mg protein) at 40 degrees C and pH 7.4. SDS-PAGE and Western blot were used to confirm nitrilase display. Compared with the free enzyme, the sporeimmobilized nitrilase showed a higher tolerance for adverse environmental conditions. After the reaction, recombinant spores were recovered via centrifugation and reused 3 times to catalyze the conversion of 3-cyanopyridine with 75.3% nitrilase activity. This study demonstrates an effective means of nitrilase immobilization via spore surface display, which can be applied in biological processes or conversion. (C) 2015 S. Karger AG, Basel

KeywordNitrilase Niacin Bacillus Subtilis Clostridium Thermocellum Spore Surface 3-cyanopyridine
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
DOI10.1159/000441642
Indexed BySCI
Language英语
WOS KeywordENZYMES ; CLONING ; SYSTEM ; ACID
WOS Research AreaBiotechnology & Applied Microbiology ; Microbiology
WOS SubjectBiotechnology & Applied Microbiology ; Microbiology
Funding OrganizationOpen Funding Project of the National Key Laboratory of Biochemical Engineering in China ; National KeyBasic Research Program of China (973 Program)(2011CBA00800) ; Key Agriculture Support Project of Jiangsu Province, China(BE2013400)
WOS IDWOS:000370260200003
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/20210
Collection管理及支撑部门
Affiliation1.Jiangsu Univ, Inst Life Sci, 301 Xuefu Rd, Zhenjiang, Peoples R China
2.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing, Peoples R China
Recommended Citation
GB/T 7714
Chen, Huayou,Zhang, Tianxi,Sun, Tengyun,et al. Clostridium thermocellum Nitrilase Expression and Surface Display on Bacillus subtilis Spores[J]. JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY,2015,25(6):381-387.
APA Chen, Huayou.,Zhang, Tianxi.,Sun, Tengyun.,Ni, Zhong.,Le, Yilin.,...&Zhang, Chunxia.(2015).Clostridium thermocellum Nitrilase Expression and Surface Display on Bacillus subtilis Spores.JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY,25(6),381-387.
MLA Chen, Huayou,et al."Clostridium thermocellum Nitrilase Expression and Surface Display on Bacillus subtilis Spores".JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY 25.6(2015):381-387.
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