Construction and co-expression of polycistronic plasmids encoding bio-degumming-related enzymes to improve the degumming process of ramie fibres
Construction and co-expression of polycistronic plasmids encoding bio-degumming-related enzymes to improve the degumming process of ramie fibres
复制标题
DOI：
10.1007/s10529-016-2204-2
复制
发表时间：
2016-12-01
影响因子：
2.7
通讯作者：
Wang, Ruijun
中科院分区：
文献类型：
--
作者：
Cheng, Yi;Liu, Zhengchu;Wang, Ruijun
为了研究三种脱胶相关酶共表达的内在特性，选育出更强大的脱胶菌株，在大肠杆菌BL21(DE3)中共表达并鉴定了脱胶过程中必需的果胶裂解酶基因、木聚糖酶基因和内切-1，4-β-甘露聚糖酶基因的6个串联多聚体。当多聚体中的XYL基因被XY91基因取代时，XY91基因与DCE-01中的内切-1，4-β-甘露聚糖酶和果胶裂解酶具有协同作用。将重组pET-PXM(91X)转化原始脱胶菌株DCE-01，提高了脱胶酶活力。此外，新型工程菌pET-PXM(91X)/DCE-01处理的样品失重率、还原糖和COD值分别提高到22.5%、460mgml(-1)和4.9，脱胶相关酶基因的共表达可应用于工业试验，为生物脱胶研究提供了新的方向。
To research the inherent properties of the co-expression of three types degumming-related enzymes and breed more powerful degumming strains.Six tandem multimers of the pectate lyase gene, the xylanase gene, and the endo-1,4-beta-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-beta-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.The co-expression of degumming-related enzyme genes may be applied in industrial tests and represents a novel direction for bio-degumming research.