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Engineering a lysine-ON riboswitch for metabolic control of lysine production in corynebacterium glutamicum
Publikationstyp
Journal Article
Date Issued
2015-09-01
Sprache
English
Author(s)
Institut
TORE-URI
Journal
Volume
4
Issue
12
Start Page
1335
End Page
1340
Citation
ACS Synthetic Biology 12 (4): 1335-1340 (2015-12-18)
Publisher DOI
Scopus ID
Publisher
ACS
Riboswitches are natural RNA elements that regulate gene expression by binding a ligand. Here, we demonstrate the possibility of altering a natural lysine-OFF riboswitch from Eschericia coli (ECRS) to a synthetic lysine-ON riboswitch and using it for metabolic control. To this end, a lysine-ON riboswitch library was constructed using tetA-based dual genetic selection. After screening the library, the functionality of the selected lysine-ON riboswitches was examined using a report gene, lacZ. Selected lysine-ON riboswitches were introduced into the lysE gene (encoding a lysine transport protein) of Corynebacterium glutamicum and used to achieve dynamic control of lysine transport in a recombinant lysine-producing strain, C. glutamicum LPECRS, which bears a deregulated aspartokinase and a lysine-OFF riboswitch for dynamic control of the enzyme citrate synthase. Batch fermentation results of the strains showed that the C. glutamicum LPECRS strain with an additional lysine-ON riboswitch for the control of lysE achieved a 21% increase in the yield of lysine compared to that of the C. glutamicum LPECRS strain and even a 89% increase in yield compared to that of the strain with deregulated aspartokinase. This work provides a useful approach to generate lysine-ON riboswitches for C. glutamicum metabolic engineering and demonstrates for the first time a synergetic effect of lysine-ON and -OFF riboswitches for improving lysine production in this industrially important microorganism. The approach can be used to dynamically control other genes and can be applied to other microorganisms.
Subjects
Corynebacterium glutamicum
lysine production
metabolic control
synthetic lysine-ON riboswitch
tetA dual genetic selection
DDC Class
570: Biowissenschaften, Biologie
600: Technik