| 黑曲霉耐热木聚糖酶XYNB在毕赤酵母中的高效表达 |
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| 引用本文： | 李伟,何容,高志强,于荣清,赵锐,曹毅,乔代蓉.黑曲霉耐热木聚糖酶XYNB在毕赤酵母中的高效表达[J].应用与环境生物学报,2011(6):892-896. |
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| 作者姓名： | 李伟 何容 高志强 于荣清 赵锐 曹毅 乔代蓉 |
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| 作者单位： | 四川大学生命科学学院微生物与代谢工程四川省重点实验室; |
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| 基金项目： | 四川省科技支撑计划项目(No.2008GZ0245); 科技人员服务企业行动项目(No.2009GJF00033)资助~~ |
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| 摘 要： | 高比活木聚糖酶的高效表达是进一步提高木聚糖酶发酵效价、降低生产成本的有效途径.将黑曲霉木聚糖酶基因XynB(不含信号肽)克隆到分泌型表达载体pPIC9K上,线性化后电击转化巴斯德毕赤酵母GS115,G418和PCR鉴定的阳性转化子经0.5%甲醇、在28℃诱导表达.SDS-PAGE分析表明,该蛋白相对分子质量为20×103左右.优化的诱导表达条件为,每隔12 h添加0.5%的甲醇,发酵5 d后,比活达4 757 U/mg;其最适温度为55℃,最适pH为5.0,80℃处理30min后仍有74%的残余酶活.
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| 关 键 词： | 黑曲霉 木聚糖酶 毕赤酵母GS115 甲醇 诱导表达 |
High Expression of Thermostable Xylanase XYNB from Aspergillus niger in Pichia pastoris |
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| LI Wei,HE Rong,GAO Zhiqiang,YU Rongqin,ZHAO Rui,CAO Yi & QIAO Dairong.High Expression of Thermostable Xylanase XYNB from Aspergillus niger in Pichia pastoris[J].Chinese Journal of Applied and Environmental Biology,2011(6):892-896. |
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| Authors: | LI Wei HE Rong GAO Zhiqiang YU Rongqin ZHAO Rui CAO Yi & QIAO Dairong |
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| Institution: | LI Wei,HE Rong,GAO Zhiqiang,YU Rongqin,ZHAO Rui,CAO Yi & QIAO Dairong(Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province,College of Life Sciences,Sichuan University,Chengdu 610064,China) |
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| Abstract: | The high expression of xylanase with high specific activity is the effective way to further improve the fermentation potency of xylanase and cut down the production costs. In this study, a recombinant plasmid pPIC9K-XynB was constructed by inserting gene XynB (without signal peptide) from Aspergillus niger into Pichia pastoris secretary vector pPIC9K. Linearized pPIC9K-XynB was transformed into P. pastoris GS115 by electroporation. The positive recombinant strain identified by G418 selection and confirmed b... |
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| Keywords: | Aspergillus niger xylanase P pastoris GS115 methanol induction expression |
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