熊海容,汪斌,杨青,张莉.不同启动子调控的两种半纤维素酶分泌表达的比较[J].中南民族大学学报自然科学版,2019,38(1):39-44
不同启动子调控的两种半纤维素酶分泌表达的比较
Comparision between two hemicellulase in secretory expression under regulation of different promoters
  
DOI:10.12130/znmdzk.20190107
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:湖北省技术创新专项重大项目(2018ABA093)
作者单位
熊海容,汪斌,杨青,张莉 中南民族大学 生命科学学院武汉 430074 
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中文摘要:
      为提高耐热木聚糖酶DSB和耐热甘露聚糖酶ManB在毕赤酵母GS115中分泌表达的酶活力,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:PFLD1, PTEF1, PGAP和PAOX1能有效介导DSB和ManB的分泌,但其胞外酶活相差较大.对于DSB,使用PAOX1时酶活力明显高于PFLD1, PTEF1和PGAP,最高酶活达846 U/mL;对于ManB,使用PFLD1时酶活力高于PAOX1,最高酶活达222 U/mL.故在GS115中分泌表达DSB时选用PAOX1,而分泌表达ManB时选用PFLD1.
英文摘要:
      To improve the expression of thermostable xylanase DSB and thermostable mannanase ManB in Pichia pastoris GS115, three strong promoters PFLD1,PGAP and PTEF1 were chosen and used to regulate the expression of DSB and ManB, using PAOX1 as a control. The shake-flask fermentation results indicated that PFLD1, PTEF1, PGAP and PAOX1 could efficiently control the secretion of DSB and ManB, but their extracellular enzyme activity differed greatly. As for DSB, the enzyme activity of PAOX1 was much higher than PFLD1, PTEF1 and PGAP,whose highest enzyme activity reached 846 U/mL. But as for ManB, the enzyme activity 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.
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