Enhanced overexpression of secreted enzymes by discrete repeat promoters in Streptomyces lividans

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Abstract
Streptomyces lividans is an efficient host for extracellular overproduction of recombinant proteins. To enhance the overexpression strength of S. lividans, we designed several kinds of expression plasmid with different positioning of repeat promoters. The effect of repeat promoters was evaluated by measuring the accumulated amounts of a stable transglutaminase or an unstable carboxypeptidase that was secreted into the medium. Successive tandem position of repeat promoter upstream the normal promoter did not enhance the expression of transglutaminase. Discrete position of repeat promoters both upstream and downstream the normal promoter enhanced the expression of transglutaminase to two-fold; and the downstream ones also enhanced the expression of carboxypeptidase to 1.7-fold. On the hand, there were still some constructs of plasmids with discrete repeat promoters that did not promote the expression of the target enzymes, indicating the complexity of the mechanisms of repeat promoters working on gene expression.
Funding Information
  • Japan Society for the Promotion of Science (26420805)

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