Activity Enhancement Based on the Chemical Equilibrium of Multiple-Subunit Nitrile Hydratase from Bordetella petrii

• Published in: Applied biochemistry and biotechnology 2016-09, Vol.180 (1), p.3-9
• Main Authors: Liu, Yi, Liu, Ping, Lin, Lu, Zhao, Yueqin, Zhong, Wenjuan, Wu, Lunjie, Zhou, Zhemin, Sun, Weifeng
• Format: Article
• Language: English
• Subjects: Bacterial Proteins - metabolism; Biochemistry; Biotechnology; Bordetella - enzymology; Cellular biology; Chemistry; Chemistry and Materials Science; Chromatography, Affinity; Electrophoresis, Polyacrylamide Gel; Enzymes; Hydro-Lyases - isolation & purification; Hydro-Lyases - metabolism; Kinetics; Models, Biological; Plasmids - metabolism; Protein Subunits - metabolism; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-016-2079-7

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, α 2 β 2 , 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.