Native promoters of Corynebacterium glutamicum and its application in l-lysine production
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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.
Compliance with ethical standards
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.