|Publisher DOI:||10.1007/s10529-017-2297-2||Title:||Reengineering substrate specificity of E. coli glutamate dehydrogenase using a position-based prediction method||Language:||English||Authors:||Geng, Feng
|Keywords:||Enzyme design;Glutamate dehydrogenase;Homoserine;Position-based prediction method;1,3-Propanediol;Protein engineering;Substrate specificity||Issue Date:||9-Feb-2017||Publisher:||Springer Science + Business Media B.V||Source:||Biotechnology Letters 4 (39): 599-605 (2017-04-01)||Journal or Series Name:||Biotechnology letters||Abstract (english):||Objective: To re-engineer the active site of proteins for non-natural substrates using a position-based prediction method (PBPM). Results: The approach has been applied to re-engineer the E. coli glutamate dehydrogenase to alter its substrate from glutamate to homoserine for a de novo 1,3-propanediol biosynthetic pathway. After identification of key residues that determine the substrate specificity, residue K92 was selected as a candidate site for mutation. Among the three mutations (K92V, K92C, and K92M) suggested by PBPM, the specific activity of the best mutant (K92 V) was increased from 171 ± 35 to 1328 ± 71 μU mg−1. Conclusion: The PBPM approach has a high efficiency for re-engineering the substrate specificity of natural enzymes for new substrates.||URI:||http://hdl.handle.net/11420/3232||ISSN:||1573-6776||Institute:||Bioprozess- und Biosystemtechnik V-1||Type:||(wissenschaftlicher) Artikel|
|Appears in Collections:||Publications without fulltext|
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