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- Title
Addition of Co(2+) to culture medium decides the functional expression of a recombinant nitrile hydratase in Escherichia coli.
- Authors
Pei, Xiaolin; Wang, Qiuyan; Li, Chenglu; Yin, Xiaopu; Chen, Rong; Xie, Tian
- 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 Co(2+) at 0.1 mM to LB medium was essential for production of an active enzyme. However, ≥0.3 mM Co(2+) inhibited the growth of host cells in LB medium and decreased the production of the recombinant NHase. Furthermore, β-mercaptoethanol promoted regeneration of the Co(2+)-defective apoenzyme in vitro possibly by breaking a key disulfide bond thereby promoting the incorporation of Co(2+) into the apoenzyme.
- Publication
Biotechnology letters, 2013, Vol 35, Issue 9, p1419
- ISSN
1573-6776
- Publication type
Journal Article
- DOI
10.1007/s10529-013-1215-5