Native promoters of Corynebacterium glutamicum and its application in l-lysine production
To identify useful native promoters of Corynebacterium glutamicum for fine-tuning of gene expression in metabolic engineering.
Sixteen native promoters of C. glutamicum were characterized. These promoters covered a strength range of 31-fold with small increments and exhibited relatively stable activity during the whole growth phase using β-galactosidase as the reporter. The mRNA level and enzymatic activity of the lacZ reporter gene exhibited high correlation (R 2 = 0.96) under the control of these promoters. Sequence analysis found that strong promoters had high similarity of the -10 hexamer to the consensus sequence and preference of the AT-rich UP element upstream the -35 region. To test the utility of the promoter library, the characterized native promoters were applied to modulate the sucCD-encoded succinyl-CoA synthetase expression for l-lysine overproduction.
The native promoters with various strengths realize the efficient and precise regulation of gene expression in metabolic engineering of C. glutamicum.
KeywordsCorynebacterium glutamicum Gene expression l-Lysine production Metabolic engineering Native promoters Promoters
This work was supported by grants from National Hi-Tech Research and Development Program of China (2014AA021203), the Science and Technology Service Network Initiative of the Chinese Academy of Sciences (KFJ-EW-STS-078) and National Natural Science Foundation of China (31100074).
Supplementary Table 1—Strains and plasmids used.
Supplementary Table 2—Primers used.
Supplementary Fig. 1—Sequence analysis of the sixteen promoters.
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Conflict of interest
The authors declare that they have no conflict of interest.
Research involving human and animal participants
The research performed did not involve human participants and/or animals.