The authors have no conflicts of interest to declare.
Isolation of fully synthetic promoters for high-level gene expression in Corynebacterium glutamicum
Article first published online: 3 JUN 2013
© 2013 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 110, Issue 11, pages 2959–2969, November 2013
How to Cite
Yim, S. S., An, S. J., Kang, M., Lee, J. and Jeong, K. J. (2013), Isolation of fully synthetic promoters for high-level gene expression in Corynebacterium glutamicum. Biotechnol. Bioeng., 110: 2959–2969. doi: 10.1002/bit.24954
- Issue published online: 21 SEP 2013
- Article first published online: 3 JUN 2013
- Accepted manuscript online: 1 MAY 2013 05:15AM EST
- Manuscript Accepted: 22 APR 2013
- Manuscript Revised: 26 MAR 2013
- Manuscript Received: 9 JAN 2013
- Ministry of Education, Science and Technology of Korea. Grant Number: ABC-2011-0031363
- Brain Korea 21 Project
- Corynebacterium glutamicum;
- synthetic promoter;
Corynebacterium glutamicum is an important industrial organism that is widely used in the production of amino acids, nucleotides and vitamins. To extend its product spectrum and improve productivity, C. glutamicum needs to undergo further engineering, including the development of applicable promoter system. Here, we isolated new promoters from the fully synthetic promoter library consisting of 70-bp random sequences in C. glutamicum. Using green fluorescent protein (GFP) as a reporter, highly fluorescent cells were screened from the library by fluorescent activated cell sorting (FACS). Twenty potential promoters of various strengths were isolated and characterized through extensive analysis of DNA sequences and mRNA transcripts. Among 20 promoters, 6 promoters which have different strengths were selected and their activities were successfully demonstrated using two model proteins (antibody fragment and endoxylanase). Finally, the strongest promoter (PH36) was employed for the secretory production of endoxylanase in fed-batch cultivation, achieving production levels of 746 mg/L in culture supernatant. This is the first report of synthetic promoters constructed in C. glutamicum, and our screening strategy together with the use of synthetic promoters of various strengths will contribute to the future engineering of C. glutamicum. Biotechnol. Bioeng. 2013;110: 2959–2969. © 2013 Wiley Periodicals, Inc.