| 甘露聚糖酶基因在毕赤酵母中的表达及酶学性质研究 |
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| 引用本文： | 乔宇陈小兵丁宏标岳明. 甘露聚糖酶基因在毕赤酵母中的表达及酶学性质研究[J]. 中国生物工程杂志, 2006, 26(7): 52-56 |
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| 作者姓名： | 乔宇陈小兵丁宏标岳明 |
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| 作者单位： | 中国农业科学院饲料研究所513室中国农业科学院饲料研究所513室中国农业科学院饲料研究所中国农业科学院饲料研究所 |
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| 基金项目： | 国家高技术研究发展计划(863计划);国家高技术研究发展计划(863计划);国际科学基金(IFS) |
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| 摘 要： | 采用PCR的方法，以枯草芽孢杆菌Bacillus subtilis基因组 DNA 为模板，克隆出甘露聚糖酶MAN的成熟肽编码序列，将其插入巴斯德毕赤酵母Pichia pastoris表达载体pPIC9K中，并位于α-因子信号肽序列的下游，获得重组质粒pPIC9K-MAN。重组质粒线性化后用聚乙二醇法导入毕赤酵母Pichia pastoris菌株GS115中，经大量筛选，获得高效分泌表达甘露聚糖酶的毕赤酵母工程菌株MAN22。将此菌株在5 L发酵罐中进行高密度发酵，测定酶活最高达1102IU /mL，同时对重组甘露聚糖酶的性质进行了初步研究。
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| 关 键 词： | 表达 甘露聚糖酶 毕赤酵母 |
| 收稿时间： | 2006-02-20 |
| 修稿时间： | 2006-04-04 |
SECRETED EXPRESSION OF MANNANASE GENE IN PICHIA PASTORIS AND ANYLYSIS OF ENZYMIC PROPERTIES |
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| QIAO Yu, CHEN Xiao-bing, DING Hong-biao, YUE Ming. SECRETED EXPRESSION OF MANNANASE GENE IN PICHIA PASTORIS AND ANYLYSIS OF ENZYMIC PROPERTIES[J]. China Biotechnology, 2006, 26(7): 52-56 |
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| Authors: | QIAO Yu CHEN Xiao-bing DING Hong-biao YUE Ming |
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| Affiliation: | Feed Research Institute, Chinese Academy of Agricultural Sciences Beijing 100081, China |
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| Abstract: | In this study a PCR method was used to amplify the sequence encoding the mature peptide of β-mannanase of Bacillus subtilis. The gene was inserted into the Pichia pastoris vector pPIC9K, downstream of α-factor signal peptide sequence. The resultant recombinant plasmid pPIC9K-MAN was lineared by BglII digestion and introduced into the host Pichia pastoris GS115 by PEG method. After screen, the recombinant P. pastoris strain MAN22 was obtained and fermented in large scale 5L fermenter. The recombinant mannanase activity could reach to 1102IU /mL . The properties of the recombinant mannanase were characterized. |
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| Keywords: | Mannanase Pichia pastoris Expression |
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