| 引用本文: | 龙传南,蒋凤姣,邬小兵,刘健,龙敏南.东方肉座菌EU7-22木聚糖酶和木糖苷酶基因克隆及生物信息学研究[J].生物信息学,2013,11(1):58-64. |
| LONG Chuan-nan,JIANG Feng-jiao,WU Xiao-bing,LIU Jian,LONG Min-nan.Gene cloning and bioinformatics analysis of Xylanases and Xylosidase from Hypocrea orientalis[J].Chinese Journal of Bioinformatics,2013,11(1):58-64. |
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| 摘要: |
| 东方肉座菌EU7-22具有高产半纤维素酶的能力。根据已报道的同属里氏木霉及绿色木霉木聚糖酶,木糖苷酶相关基因序列,设计PCR引物扩增出东方肉座菌内切木聚糖酶(XYNⅠ,XYNⅡ)及β-木糖苷酶(β-BXL)基因。序列经NCBI Blast分析:东方肉座菌xynⅠ基因与里氏木霉xyn1基因(X69573.1)的同源性最高达到91%；xyn Ⅱ基因与绿色木霉xyn2基因(EF079061)同源性最高达到93%；β-bxl 基因与里氏木霉β-bxl1基因(Z69257.1)的同源性最高达到94%。生物信息学分析表明内切木聚糖酶Ⅰ和Ⅱ均属于糖基水解酶家族11,N末端前19个氨基酸均为信号肽。内切木聚糖酶Ⅰ分子量为24.13kD,等电点为7.87,含有3个糖基化位点；内切木聚糖酶Ⅱ分子量为24.44kD,等电点为4.86,含有1个糖基化位点；β-木糖苷酶属于糖基水解酶家族3,分子量为87.27kD,等电点为5.49,N 末端前20个氨基酸为信号肽,含有8个糖基化位点。利用SWISS-Model对木聚糖酶,木糖苷酶蛋白质三级结构进行了预测和模拟。对木聚糖酶和木糖苷酶基因及其编码蛋白质的生物信息学分析,为进一步研究这些基因的表达与调控、构建高效利用纤维素组份的工程菌株奠定基础。 |
| 关键词: 东方肉座菌,木聚糖酶,木糖苷酶,基因克隆,生物信息学 |
| DOI：10.3969/j.issn.1672-5565.2013-01.20130110 |
| 分类号: |
| 基金项目:国家重点基础研究发展计划(973计划)(NO.2010CB732201); 中央高校基本科研业务费专项资金(NO.201112G026); 国家自然科学基金(NO. 31170067)。 |
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| Gene cloning and bioinformatics analysis of Xylanases and Xylosidase from Hypocrea orientalis |
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LONG Chuan-nan1,JIANG Feng-jiao1,WU Xiao-bing2,LIU Jian1,LONG Min-nan1,2
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(1. School of Energy Research, Xiamen University, Xiamen Fujian 361005, China; ;2. School of Life Sciences, Xiamen University, Xiamen Fujian 361005, China)
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| Abstract: |
| Hypocrea orientalis EU7-22had a high potential to yield hemicellulase. Two endo-xylanases genes (xyn Ⅰ, xyn Ⅱ) and one β-xylosidase gene (β-bxl) were successfully cloned by PCR, according to the reported xylanases and β-xylosidase gene sequences of Trichoderma reesei and Trichoderma viride. The xyn I and β-bxl gene from H. orientalis showed the highest nucleotide homology of 91% and 94% with the corresponding gene from T. reesei, respectively. While the xyn II gene from H. orientalis showed the highest of 93% nucleotide homology with the corresponding gene from T. viride. The bioinformatics analysis indicated that the enzyme XYN I and XYN II belonged to the glycosyl hydrolase family 11with the molecular weight of 24.13kD and 24.44kD, respectively. The first 19AA of N-terminal of XYN I and XYN II were the signal peptide sequence. The enzyme XYN I and XYN II contained three and one N-glycosylation site, respectively. The isoelectric point of enzyme XYN I and XYN II were identified as 7.87and 4.86, respectively. The enzyme β-BXL belonged to glycosyl hydrolase family 3with molecular weight of 87.27kD and isoelectric point of 5.49. The first 20AA of N-terminal of β-BXL belonged to signal peptide sequence. The enzyme contained 8N-glycosylation sites. By using SWISS-Model, the tertiary structure of the three enzyme proteins were predicted and simulated. These genes cloning and bioinformatics analysis would help to the further research on mechanism of expression and regulation of hemicellulase. |
| Key words: Hypocrea orientalis,Xylanases, Xylosidase, Gene cloning, Bioinformatics |
