1. Computer-aided engineering of adipyl-CoA synthetase for enhancing adipic acid synthesis.
- Author
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Yang J, Wei Y, Li G, Zhou S, and Deng Y
- Subjects
- Adipates chemistry, Catalysis, Cephalosporins pharmacology, Escherichia coli genetics, Hydrolases chemistry, Ligases chemistry, Mutagenesis, Site-Directed, Substrate Specificity, Adipates metabolism, Computer-Aided Design, Hydrolases biosynthesis, Ligases biosynthesis
- Abstract
Objective: To enhance adipic acid production, a computer-aided approach was employed to engineer the adipyl-CoA synthetase from Thermobifida fusca by combining sequence analysis, protein structure modeling, in silico site-directed mutagenesis, and molecular dynamics simulation., Results: Two single mutants of T. fusca adipyl-CoA synthetase, E210βN and E210βQ, achieved a specific enzyme activity of 1.95 and 1.84 U/mg, respectively, which compared favorably with the 1.48 U/mg for the wild-type. The laboratory-level fermentation experiments showed that E210βN and E210βQ achieved a maximum adipic acid titer of 0.32 and 0.3 g/L. In contrast, the wild-type enzyme yielded a titer of 0.15 g/L under the same conditions. Molecular dynamics (MD) simulations revealed that the mutants (E210βN and E210βQ) could accelerate the dephosphorylation process in catalysis and enhance enzyme activity., Conclusions: The combined computational-experimental approach provides an effective strategy for enhancing enzymatic characteristics, and the mutants may find a useful application for producing adipic acid.
- Published
- 2020
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