| 热稳定高比活纤维素酶在Pichia pastoris中的表达研究 |
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| 引用本文： | 江均平,蔡丽君,龚霄,张杰,魏益民.热稳定高比活纤维素酶在Pichia pastoris中的表达研究[J].工业微生物,2010,40(5):27-31. |
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| 作者姓名： | 江均平 蔡丽君 龚霄 张杰 魏益民 |
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| 作者单位： | 中国农业科学院农产品加工研究所,北京,100094 |
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| 基金项目： | 北京市自然科学基金项目，国家高技术研究发展计划，中国农业科学院科技经费项目 |
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| 摘 要： | Bacillus amyoliquefaciens DL-3纤维素酶具有热稳定高比活多功能的酶学特性,本文根据该酶的氨基酸序列合成了其编码基因(cel),构建了pPIC9K-cel表达载体,并用P.pastoris进行了表达。工程菌株三角瓶发酵酶活性达0.50 U/mL,酶解滤纸的产物为低聚糖,表明人工合成的热稳定高比活纤维素酶基因在P.pastoris中可以正常表达、加工及分泌,重组酶的分子量由天然酶的53 kDa增加至68 kDa,糖基化严重。
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| 关 键 词： | 纤维素酶 解淀粉芽胞杆菌 毕赤酵母 糖基化 |
Expression of thermostable cellulase with high specific activity from Bacillus amyoliquefaciens DL-3 in Pichia pastoris |
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| JIANG Jun-ping,Cai Li-jun,Gong Xiao,Zhang Jie,Wei Yi-min.Expression of thermostable cellulase with high specific activity from Bacillus amyoliquefaciens DL-3 in Pichia pastoris[J].Industrial Microbiology,2010,40(5):27-31. |
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| Authors: | JIANG Jun-ping Cai Li-jun Gong Xiao Zhang Jie Wei Yi-min |
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| Institution: | ( Institute of Agro-Fcxxt Science and Tec｜moiogy, Chinese Academy of Agricultural Sciences, Beijing 100094, China ) |
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| Abstract: | A complete gene was synthesized according to the amino acid sequence of Bacillus amyoliquefaciens DL-3 cellulase and expressed extracellularly in Pichia pastoris. The CMC activity of the recombinant P. pastoris strain reached 0.5 U/mL in Erlenmeyer flask, and the hydrolytic products of paper by the recombined cellulase were oligosaccharides. The molecular mass of the recombinant enzyme was found to be about 68 kDa by SDS PAGE, which was much higher than 53 kDa of the nature one. Glycosylation was presumed to be the cause of low activity. |
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| Keywords: | cellulase Bacillus amyoliquefaciens Pichia pastoris glycosilation |
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