不同启动子调控的两种半纤维素酶分泌表达的比较
Extracellular Expression of Two Hemicellulase in Pichia pastoris under the Regulation of Different Promoters
投稿时间:2017-01-21  修订日期:2017-03-13
DOI:
中文关键词: 启动子  巴斯德毕赤酵母  分泌表达  半纤维素酶
英文关键词: Promoter  Pichia pastoris  Secretory expression  Hemicellulase
基金项目:中央高校基本科研业务费专项资金项目
作者单位E-mail
熊海容 中南民族大学生命科学学院 xionghr@mail.scuec.edu.cn 
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中文摘要:
      木聚糖酶和甘露聚糖酶是两种主要的半纤维素酶,广泛应用于诸多领域。为提高实验室前期获得的耐热木聚糖酶DSB和耐热甘露聚糖酶ManB在毕赤酵母GS115中的表达量,本研究选择了3个调控蛋白表达能力较强的启动子PFLD1、PGAP和PTEF1,以PAOX1为参照,分别在毕赤酵母GS115中进行分泌表达。摇瓶发酵结果表明不同启动子调控的两种半纤维素酶的分泌表达水平相差较大:对于DSB,使用PAOX1时酶活力明显高于其它启动子,最高酶活达846 U/ml;对于ManB,使用P FLD1时酶活力高于PAOX1,最高酶活达222 U/ml。因此在GS115中分泌表达DSB时应选用PAOX1,而分泌表达ManB时应选用P FLD1。本研究为实现半纤维素酶在毕赤酵母中的高效表达奠定了一定的基础。
英文摘要:
      Xylanase and mannanase are two hemicellulases which are widely used in many fields. In order to improve the expression of thermostable xylanase DSB and thermostable mannanase ManB in Pichia pastoris GS115, three strong promoter PFLD1, PTEF and PGAP were chosed and used to regulate the expression of DSB and ManB. PAOX1 was used as a control. In shake-flask cultivation, PFLD1, PTEF, PGAP and PAOX1 efficiently control the secretion of DSB and ManB, but the expression efficiency has obvious difference. As for DSB, the expression efficiency of PAOX1 was much higher than PFLD1, PTEF and PGAP and the highest enzyme activity reached 846 U/ml. But as for ManB, the expression efficiency of PFLD1 was higher than PAOX1 and the highest enzyme activity reached 222 U/ml. Therefore, PAOX1 was the most efficient promoter for DSB expression and PFLD1 was the most efficient promoter for ManB expression in Pichia pastoris GS115, respectively. This study laid a foundation for increasing the expression of hemicellulase in Pichia pastoris.
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