Reengineering substrate specificity of E. coli glutamate dehydrogenase using a position-based prediction method

Publikationstyp
Journal Article
Date Issued
2017-02-09
Sprache
English
Author(s)
Geng, Feng  
Ma, Cheng-Wei  
Zeng, An-Ping  orcid-logo
Institut
Bioprozess- und Biosystemtechnik V-1  
TORE-URI
http://hdl.handle.net/11420/3232
Journal
Biotechnology letters  
Volume
39
Issue
4
Start Page
599
End Page
605
Citation
Biotechnology Letters 4 (39): 599-605 (2017-04-01)
Publisher DOI
10.1007/s10529-017-2297-2
Scopus ID
2-s2.0-85011879314
Publisher
Springer Science + Business Media B.V
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.
Subjects
Enzyme design
Glutamate dehydrogenase
Homoserine
Position-based prediction method
1,3-Propanediol
Protein engineering
Substrate specificity
DDC Class
570: Biowissenschaften, Biologie
610: Medizin