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篇目详细内容 |
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【篇名】 |
Cloning of endo-β-glucanase I gene and expression in Pichia pastoris |
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【刊名】 |
Frontiers of Agriculture in China |
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【刊名缩写】 |
Front. Agric. China |
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【ISSN】 |
1673-7334 |
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【EISSN】 |
1673-744X |
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【DOI】 |
10.1007/s11703-011-1082-6 |
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【出版社】 |
Higher Education Press and Springer-Verlag Berlin
Heidelberg |
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【出版年】 |
2011 |
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【卷期】 |
5
卷2期 |
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【页码】 |
196-200
页,共
5
页 |
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【作者】 |
Yu BAI;
Runfang GUO;
Hongwei YU;
Long JIAO;
Shuli DING;
Yingmin JIA;
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【关键词】 |
Thermoascus aurantiacus; endo-1; 4-β-glucanase; RACE; Pichia pastoris; gene expression |
【摘要】 |
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Total RNA of Thermoascus aurantiacus was isolated from its mycelium and acted as template for RT-PCR. The full-length cDNA encoding an endo-β-glucanase I was cloned via RACE-PCR method and the cDNA contained an ORF of 1005？bp encoding 305 amino acids. A recombinant plasmid, pPIC9k-egI, was constructed by inserting the ORF sequence of endo-β-glucanase I gene (egI) into the yeast expression vector pPIC9k and transformed to Pichia pastoris GS115. The results showed that the recombinant endo-β-glucanase I was excreted into the fermentation medium. The highest activity of endo-β-glucanase I and the protein content were up to 45.42？U/mL and 788.26？μg/mL at incubation time of 144？h. The optimal temperature and pH for the recombinant endo-β-glucanase I were found to be 70oC and 3.5, respectively. |
