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
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国家研发计划项目(2017YFC1600605);北京自然科学基金项目(6172003);国家自然科学基金项目(31601408)


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

    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xynGH11全长729 bp;分子质量:24.003 ku;理论等电点p I: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。

    Abstract:

    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.

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熊科;熊苏玥;柳佳芸;高思宇;邓蕾;裴鹏钢;.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[J].中国食品学报,2019,19(10):56-67

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  • 在线发布日期: 2019-11-21
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