Development of bicistronic expression system for the enhanced and reliable production of recombinant proteins in Leuconostoc citreum

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The lactic acid bacteria (LAB) Leuconostoc citreum are non-sporulating hetero-fermentative bacteria that play an important role in the fermented food industry. In this study, for the enhanced and reliable production of recombinant proteins in L. citreum, we developed a bicistronic design (BCD) expression system which includes a short leader peptide (1st cistron) followed by target genes (2nd cistron) under the control of a single promoter. Using superfolder green fluorescent protein (sfGFP) as a reporter, the functionality of BCD in L. citreum was verified. Further, to improve the expression in BCD, we tried to engineer a Shine-Dalgarno sequence (SD2) for the 2nd cistron and a promoter by FACS screening of random libraries, and both strong SD2 (eSD2) and promoter (P-710V4) were successfully isolated. The usefulness of the engineered BCD with P-710V4 and eSD2 was further validated using three model proteins-glutathione-s-transferase, human growth hormone, and alpha-amylase. All examined proteins were successfully produced with levels highly increased compared with those in the original BCD as well as the monocistronic design (MCD) expression system.
Publisher
NATURE PUBLISHING GROUP
Issue Date
2018-06
Language
English
Article Type
Article
Citation

SCIENTIFIC REPORTS, v.8, pp.8852

ISSN
2045-2322
DOI
10.1038/s41598-018-27091-z
URI
http://hdl.handle.net/10203/244031
Appears in Collection
CBE-Journal Papers(저널논문)
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