Options
Exploring the allosteric mechanism of dihydrodipicolinate synthase by reverse engineering of the allosteric inhibitor binding sites and its application for lysine production
Publikationstyp
Journal Article
Date Issued
2012-05-29
Sprache
English
Author(s)
Institut
TORE-URI
Volume
97
Issue
5
Start Page
1963
End Page
1971
Citation
Applied Microbiology and Biotechnology 5 (97): 1963-1971 (2013)
Publisher DOI
Scopus ID
Publisher
Springer
Dihydrodipicolinate synthase (DHDPS, EC 4.2.1.52) catalyzes the first committed reaction of l-lysine biosynthesis in bacteria and plants and is allosterically regulated by l-lysine. In previous studies, DHDPSs from different species were proved to have different sensitivity to l-lysine inhibition. In this study, we investigated the key determinants of feedback regulation between two industrially important DHDPSs, the l-lysine-sensitive DHDPS from Escherichia coli and l-lysine-insensitive DHDPS from Corynebacterium glutamicum, by sequence and structure comparisons and site-directed mutation. Feedback inhibition of E. coli DHDPS was successfully alleviated after substitution of the residues around the inhibitor's binding sites with those of C. glutamicum DHDPS. Interestingly, mutagenesis of the lysine binding sites of C. glutamicum DHDPS according to E. coli DHDPS did not recover the expected feedback inhibition but an activation of DHDPS by l-lysine, probably due to differences in the allosteic signal transduction in the DHDPS of these two organisms. Overexpression of l-lysine-insensitive E. coli DHDPS mutants in E. coli MG1655 resulted in an improvement of l-lysine production yield by 46 %. © 2012 Springer-Verlag.
Subjects
Allosteric binding site
Corynebacterium glutamicum
Dihydrodipicolinate synthase (DHDPS)
Escherichia coli
l-Lysine overproduction
DDC Class
570: Biowissenschaften, Biologie
More Funding Information
Chinese Academy of Sciences (project KSCX2-EW-G-14-1)
Deutsche Forschungsgemeinschaft (DFG) (project ZE 542/6-1)