Construction and co-expression of polycistronic plasmids encoding bio-degumming-related enzymes to improve the degumming process of ramie fibres

Biotechnol Lett. 2016 Dec;38(12):2089-2096. doi: 10.1007/s10529-016-2204-2. Epub 2016 Sep 15.

Abstract

Objectives: To research the inherent properties of the co-expression of three types degumming-related enzymes and breed more powerful degumming strains.

Results: Six tandem multimers of the pectate lyase gene, the xylanase gene, and the endo-1,4-β-mannanase gene, which are essential for degumming process, were co-expressed and evaluated in Escherichia coli BL21(DE3). The xyl91 gene had a synergistic effect with endo-1,4-β-mannanase and pectate lyase from DCE-01, when xyl gene was replaced with xyl91 in the multimer. The recombinant pET-pxm(91x) was selected and transformed into the original degumming strain DCE-01, which led to an enzymatic activity improvement. Furthermore, the weight loss, reducing sugar and COD value of the sample treated with the new engineered strain pET-pxm(91x)/DCE-01 increased to 22.5 %, 460 mg ml-1 and 4.9, respectively.

Conclusions: The co-expression of degumming-related enzyme genes may be applied in industrial tests and represents a novel direction for bio-degumming research.

Keywords: BE-91; Bio-degumming; Co-expression; DCE-01; Ramie; Recombinant strains; xyl91.

MeSH terms

  • Biodegradation, Environmental
  • Biotechnology / methods
  • Escherichia coli / enzymology
  • Escherichia coli / metabolism
  • Mannosidases / genetics
  • Plasmids / genetics*
  • Polysaccharide-Lyases / genetics*
  • Xylosidases / genetics*

Substances

  • Mannosidases
  • Xylosidases
  • endo-1,4-beta-D-mannanase
  • Polysaccharide-Lyases
  • pectate lyase