Chaperones-assisted soluble expression and maturation of recombinant Co-type nitrile hydratase in Escherichia coli to avoid the need for a low induction temperature

Journal of Biotechnology
Xiaolin PeiJianping Wu

Abstract

Nitrile hydratase (NHase) is an important industrial enzyme that biosynthesizes high-value amides. However, most of NHases expressed in Escherichia coli easily aggregate to inactive inclusion bodies unless the induction temperature is reduced to approximately 20°C. The NHase from Aurantimonas manganoxydans has been functionally expressed in E. coli, and exhibits considerable potential for the production of nicotinamide in industrial application. In this study, the effects of chaperones including GroEL/ES, Dnak/J-GrpE and trigger factor on the expression of the recombinant Co-type NHase were investigated. The results indicate that three chaperones can significantly promote the active expression of the recombinant NHase at 30°C. The total NHase activities reached to 263 and 155U/ml in shake flasks when the NHase was co-expressed with GroEL/ES and DnaK/J-GrpE, which were 52- and 31-fold higher than the observed activities without chaperones, respectively. This increase is possibly due to the soluble expression of the recombinant NHase assisted by molecular chaperones. Furthermore, GroEL/ES and DnaK/J-GrpE were determined to promote the maturation of the Co-type NHase in E. coli under the absence of the parental activator gene. The...Continue Reading

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Citations

May 12, 2020·Frontiers in Bioengineering and Biotechnology·Zhongyi ChengZhemin Zhou
Jun 16, 2019·Molecular Biology Reports·K SupreethaS Kiran
Jul 2, 2019·International Journal of Biological Macromolecules·Fatemeh VahdaniReza Khodarahmi
Sep 2, 2021·Journal of Agricultural and Food Chemistry·Shilei SunMiaomiao Sheng

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