Screening and application of inducible promoters in Ruminiclostridium papyrosolvens.

Abstract

Ruminiclostridium papyrosolvens is a promising candidate for producing renewable green chemicals from cellulose due to its cellulolytic and ethanologenic capabilities. It is of significance to screen effective, and convenient-to-use inducible promoters that can be used for regulating the gene expression in R. papyrosolvens. We characterized two endogenous inducible promoters and investigated another two exogenous ones on the adaptability in R. papyrosolvens. Both of the endogenous xylan-inducible promoter Pxyl and exogenous lactose-inducible promoter Plac are found of high specificity and stringency. Pxyl has a short time to be induced while Plac has a low concentration of inducer. With these findings, a mazF-based counter selectable system has been constructed for promoting the efficiency of mutant screening via plasmid curing. The inducible gene expression systems provided novel tools for enhancing the capability of genetic manipulation in engineering R. papyrosolvens. SIGNIFICANCE AND IMPACT OF THE STUDY: Four inducible promoters from Clostridia were characterized in R. papyrosolvens. Xylan-inducible promoter Pxyl was found of a short time while lactose-inducible promoter Plac needs a low concentration of inducer to induce. Employing them, we successfully construct a mazF-based counter selectable system, which would be used to increase the mutant screening efficiency via induction of plasmid curing. The inducible gene expression systems provided novel tools for enhancing the capability of genetic manipulation in engineering R. papyrosolvens.