title: The Stability Enhancement of Nitrile Hydratase from Bordetella petrii by Swapping the C-terminal Domain of β subunit.
abstract: The thermal stability of most nitrile hydratases (NHase) is poor, which has been enhanced to some extent by molecular modifications in several specific regions of the C-terminal domain (C-domain) of β subunit of NHase. Since the C-domain could be present as a naturally separate domain in a few NHases, the whole C-domain is proposed to be related to the NHase stability. The chimeric NHase (SBpNHase) from the thermal-sensitive BpNHase (NHase from Bordetella petrii) and the relatively thermal-stable PtNHase (NHase from Pseudonocardia thermophila) was constructed by swapping the corresponding C-domains. After 30 min incubation at 50 °C, the original BpNHase nearly lost its activity, while the SBpNHase retained 50 % residual activity, compared with the melting temperature (Tm) (50 °C) of the original BpNHase, that of the SBpNHase was 55 °C. The SBpNHase with higher thermal stability would be useful for the thermal stability enhancement of NHase and for the understanding of the relationship between the stability of NHase and its structure.
authors.affiliation: Key Laboratory of Food and Biotechnology, School of Food and Biotechnology, Xihua University, Chengdu, Sichuan, 610039, People's Republic of China.
authors.forename: Weifeng
authors.initials: W
authors.lastname: Sun
authors.affiliation: School of Biochemical Engineering, Anhui Polytechnic University, Anhui, 241000, China.
authors.forename: Longbao
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Food and Biotechnology, School of Food and Biotechnology, Xihua University, Chengdu, Sichuan, 610039, People's Republic of China.
authors.forename: Xianggui
authors.initials: X
authors.lastname: Chen
authors.affiliation: Key Laboratory of Food and Biotechnology, School of Food and Biotechnology, Xihua University, Chengdu, Sichuan, 610039, People's Republic of China.
authors.forename: Lunjie
authors.initials: L
authors.lastname: Wu
authors.affiliation: Key Laboratory of Industrial Biotechnology, School of Biotechnology, Jiangnan University, Wuxi, 214122, China.
authors.forename: Zhemin
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Food and Biotechnology, School of Food and Biotechnology, Xihua University, Chengdu, Sichuan, 610039, People's Republic of China. puter_scholar@163.com.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence / Q000235:genetics; D001884:Bordetella / Q000201:enzymology*; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D006836:Hydro-Lyases / Q000737:chemistry* / Q000235:genetics; D000072417:Protein Domains; D021122:Protein Subunits / Q000737:chemistry; D013696:Temperature
chemical_list: D021122:Protein Subunits; D006836:Hydro-Lyases; C048831:nitrile hydratase
keywords: Bordetella petrii; Enzyme thermal stability; NHase; Pseudonocardia thermophila