β-xylosidase analysis and expression levels of xyl gene in different strains of Volvariella volvacea
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摘要: 本研究首先对草菇β-木糖苷酶编码序列Xyl进行了生物信息学分析,然后以对硝基木糖苷为底物测定了不同菌株中的木糖苷酶活性,并进一步比较了木糖苷酶基因xyl在草菇不同菌株中的相对表达量。结果表明:Xyl属于糖苷水解酶第43家族,由534个氨基酸残基组成,预测分子量为58.82ku,等电点为4.99;系统进化树分析表明,Xyl的氨基酸序列与木腐菌相似性较高,最高值达到61%,且草菇与木腐菌黄孢原毛平革、毛韧革菌等亲缘关系较近,但与同为草腐菌的灰盖鬼伞遗传距离较远;草菇不同菌株的β-木糖苷酶活性测定结果显示,Vb1的β-木糖苷酶活性最高,0229的β-木糖苷酶活性与A8之间无显著性差异;Real-time PCR结果显示,Vb1的xyl基因相对表达量也最高,三个菌株的xyl基因相对表达量依次为Vb1>A8>0229。Abstract: This paper studied β-xylosidase activity and xyl gene expression in different strains of Volvariella volvacea. Bioinformatics traits of Xyl were analyzed. Xylosidase activities in different Volvariella volvacea strains were determined by 4-Nitrophenyl-β-D-xylopyranoside. The xyl gene expression was measured by real time-PCR using GAPDH as the reference gene. Results showed that Xyl belonged to glycoside hydrolase family 43 and consisted of 534 amino acid residues. The predicted molecular weight and isoelectric point of Xyl was 58.82 ku and 4.99, respectively. Phylogenetic analysis indicated that Xyl was similar with wood rot fungi at61% level. Xyl in Volvariella volvacea was much closer to that in wood rot fungus Phanerochaete chrysosporium or Stereum hirsutum than to the homologous sequence in grass rotting fungus Coprinopsis cinerea. Xylosidase activity in Vb1 was the highest;however, no significant difference of xylosidase activity was observed between0229 and A8. The real time-PCR analysis showed that relative expression of xyl gene in different strains of Volvariella volvacea was Vb1>A8>0229.
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Keywords:
- Volvariella volvacea /
- β-xylosidase /
- real-time PCR /
- bioinformatics
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