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Rational design of allosteric regulation of homoserine dehydrogenase by a nonnatural inhibitor l -lysine
Publikationstyp
Journal Article
Date Issued
2014-01-06
Sprache
English
Author(s)
Institut
TORE-URI
Journal
Volume
4
Issue
2
Start Page
126
End Page
131
Citation
ACS Synthetic Biology 2 (4): 126-131 (2015)
Publisher DOI
Scopus ID
Publisher
ACS
Allosteric proteins, which can sense different signals, are interesting biological parts for synthetic biology. In particular, the design of an artificial allosteric enzyme to sense an unnatural signal is both challenging and highly desired, for example, for a precise and dynamical control of fluxes of growth-essential but byproduct pathways in metabolic engineering of industrial microorganisms. In this work, we used homoserine dehydrogenase (HSDH) of Corynebacterium glutamicum, which is naturally allosterically regulated by threonine and isoleucine, as an example to demonstrate the feasibility of reengineering an allosteric enzyme to respond to an unnatural inhibitor l-lysine. For this purpose, the natural threonine binding sites of HSD were first predicted and verified by mutagenesis experiments. The threonine binding sites were then engineered to a lysine binding pocket. The reengineered HSD only responds to lysine inhibition but not to threonine. This is a significant step toward the construction of artificial molecular circuits for dynamic control of growth-essential byproduct formation pathway for lysine biosynthesis.
Subjects
allosteric regulation
dynamic flux control
homoserine dehydrogenase
lysine
protein design
DDC Class
570: Biowissenschaften, Biologie
600: Technik
More Funding Information
Funding sponsor: Deutsche Forschungsgemeinschaft (DFG) AOBJ 588373, Deutsche Forschungsgemeinschaft (DFG) ZE 542/6.