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Addition of Co2+ to culture medium decides the functional expression of a recombinant nitrile hydratase in Escherichia coli

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Abstract

A nitrile hydratase (NHase) gene from Aurantimonas manganoxydans, cloned and expressed in Escherichia coli, gave an enzyme that efficiently hydrated 3-cyanopyridine to nicotinamide with high thermal stability. We have now found that adding Co2+ at 0.1 mM to LB medium was essential for production of an active enzyme. However, ≥0.3 mM Co2+ inhibited the growth of host cells in LB medium and decreased the production of the recombinant NHase. Furthermore, β-mercaptoethanol promoted regeneration of the Co2+-defective apoenzyme in vitro possibly by breaking a key disulfide bond thereby promoting the incorporation of Co2+ into the apoenzyme.

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (No. 21206024), and Zhejiang Provincial Natural Science Foundation of China (No. LQ12B06007).

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Authors and Affiliations

  1. Center for Biomedicine and Health, College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou, 311121, China

    Xiaolin Pei, Qiuyan Wang, Chenglu Li, Xiaopu Yin, Rong Chen & Tian Xie

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  1. Xiaolin Pei
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  2. Qiuyan Wang
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  3. Chenglu Li
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  4. Xiaopu Yin
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  5. Rong Chen
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  6. Tian Xie
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Correspondence to Xiaolin Pei.

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Pei, X., Wang, Q., Li, C. et al. Addition of Co2+ to culture medium decides the functional expression of a recombinant nitrile hydratase in Escherichia coli . Biotechnol Lett 35, 1419–1424 (2013). https://doi.org/10.1007/s10529-013-1215-5

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  • DOI: https://doi.org/10.1007/s10529-013-1215-5

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