title: Activity Enhancement Based on the Chemical Equilibrium of Multiple-Subunit Nitrile Hydratase from Bordetella petrii.
abstract: The maturation mechanism of nitrile hydratase (NHase) of Pseudomonas putida NRRL-18668 was discovered and named as "self-subunit swapping." Since the NHase of Bordetella petrii DSM 12804 is similar to that of P. putida, the NHase maturation of B. petrii is proposed to be the same as that of P. putida. However, there is no further information on the application of NHase according to these findings. We successfully rapidly purified NHase and its activator through affinity his tag, and found that the cell extracts of NHase possessed multiple types of protein ingredients including α, β, α2β2, and α(P14K)2 who were in a state of chemical equilibrium. Furthermore, the activity was significantly enhanced through adding extra α(P14K)2 to the cell extracts of NHase according to the chemical equilibrium. Our findings are useful for the activity enhancement of multiple-subunit enzyme and for the first time significantly increased the NHase activity according to the chemical equilibrium.
authors.affiliation: Key Laboratory of Food and Biotechnology, School of Food and Biotechnology, Xihua University, Chengdu, 610039, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Key Laboratory of Food and Biotechnology, School of Food and Biotechnology, Xihua University, Chengdu, 610039, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Liu
authors.affiliation: Key Laboratory of Food and Biotechnology, School of Food and Biotechnology, Xihua University, Chengdu, 610039, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Lin
authors.affiliation: Key Laboratory of Food and Biotechnology, School of Food and Biotechnology, Xihua University, Chengdu, 610039, China.
authors.forename: Yueqin
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Food and Biotechnology, School of Food and Biotechnology, Xihua University, Chengdu, 610039, China.
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Zhong
authors.affiliation: Key Laboratory of Food and Biotechnology, School of Food and Biotechnology, Xihua University, Chengdu, 610039, 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, 610039, China. sunweifeng1024@163.com.
authors.forename: Weifeng
authors.initials: W
authors.lastname: Sun
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D001884:Bordetella / Q000201:enzymology*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006836:Hydro-Lyases / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008954:Models, Biological; D010957:Plasmids / Q000378:metabolism; D021122:Protein Subunits / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D021122:Protein Subunits; D011994:Recombinant Proteins; D006836:Hydro-Lyases; C048831:nitrile hydratase
keywords: Activity; Bordetella petrii; Chemical equilibrium; NHase; Self-subunit swapping