链霉菌L10608来源木聚糖酶xyn-GH10/11基因的克隆与生物信息分析

Cloning and Bioinformatics Analysis of xyn-GH10/11 Gene from Streptomyces. sp L10608

基于菌株L10608所产天然酶水解木聚糖底物时具有良好的水解木聚糖底物特性,通过设计简并引物,经巢式PCR克隆得到L10608菌株木聚糖酶基因全长(xyn-GH10与xyn-GH11),并对其进行生物信息学分析。由分析可知酶xyn-GH10全长1 469 bp,分子质量:50.677 ku;理论等电点pI:7.98;预测该木聚糖酶的信号肽为MGIQALPRAAVRQKLRTPLPALAAGVLGLTAALVPPTNADA;该木聚糖酶蛋白序列具有明显的第10族糖苷水解酶的八叠桶型保守区域,且能编码一段由108个氨基酸组成的RicinB-lectin非催化结构域。木聚糖酶xyn-GH11全长729 bp;分子质量:24.003 ku;理论等电点pI:8.98;预测该木聚糖酶的信号肽为MHQDGSQQDRT QNPAPFGGLSRRGFLVGGTGAAALAAGSGLLLPGTAHAA。该木聚糖酶蛋白序列具有明显的第11族糖苷水解酶的“右手半握状”保守区域。xyn-GH10与xyn-GH11基因经原核表达后其中木聚糖酶xyn-GH10酶活为160 U/mL,木聚糖酶xyn-GH11酶活为55 U/mL。

The xylanase produced by strain L10608 has good hydrolysis properties when hydrolyzed substrate of xylan.The xylanase gene(xyn-GH10 and xyn-GH11)of L10608 was obtained by nested PCR.The xylanase gene(xyn-GH10 and xyn-GH11)was obtained by nested PCR,then the bioinformatics analysis was performed.The xyn-GH10 has a total length of 1 469 bp,molecular weight:50.677 ku;theoretical pI:7.98;It predicted that the xylanase signal peptide is MGIQALPRAAVRQKLRTPLPALAAGVLGLTAALVPPTNADA by Signal4.0;the xylanase protein sequence has distinct family 10 glycoside hydrolase sarcina barrel conserved region catalysis domain,and encoded a non-catalytic section RicinB-lectin domain consists of 108 amino acids.xyn-GH11 full length is 729 bp;molecular weight:24.003 ku;theoretical pI:8.98;It predicted that the xylanase signal peptide is MHQDGSQQDRTQNPAPFGGLSRRGFLVGGTGAAALAAGSGLLLPGTAHAA by signal 4.0.The xylanase protein sequence has a distinct“right-hand”conserved region of the family 11 of glycoside hydrolases.The xyn-GH10 activity was 160 U/mL,and the activity of xyl-GH11 was 55 U/mL.