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Construction and co-expression of polycistronic plasmids encoding bio-degumming-related enzymes to improve the degumming process of ramie fibres
- Cheng, Yi, Liu, Zhengchu, Zeng, Jie, Cheng, Lifeng, Yan, Zhun, Duan, Shengwen, Feng, Xiangyuan, Zheng, Ke, Zheng, Xia, Wang, Ruijun
- Biotechnology letters 2016 v.38 no.12 pp. 2089-2096
- Escherichia coli, chemical oxygen demand, degumming, enzyme activity, genes, pectate lyase, plasmids, ramie, reducing sugars, synergism, weight loss, xylanases
- 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.