蚓激酶基因在毕赤酵母中的表达及其发酵条件优化

  • 江鹏 ,
  • 汤斌.
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  • 1(安徽工程大学 生物与化学工程学院,安徽 芜湖,241000) 2(微生物发酵安徽省工程研究中心,安徽 芜湖,241000)
硕士研究生

收稿日期: 2018-03-06

  修回日期: 2018-04-08

  网络出版日期: 2018-11-23

The expression of lumbrokinase gene in Pichia pastoris and the optimizationof high density fermentation conditions

  • JIANG Peng ,
  • TANG Bin.
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  • 1(School of Biological and Chemical Engineering, Anhui Polytechnic University, Wuhu 241000, China) 2(Anhui Engineering Technology Research Center of Microbial Fermentation, Wuhu 241000, China)

Received date: 2018-03-06

  Revised date: 2018-04-08

  Online published: 2018-11-23

摘要

从赤子爱胜蚓体内克隆得到蚓激酶基因lks2,并成功在毕赤酵母GS115中表达,酶活为254.4 U/mL。经摇瓶培养条件优化,确定最佳诱导条件为:初始OD600=1.5,pH 5.5、温度28 ℃,此条件下诱导84 h,重组菌GS115-pPIC9K-lks2蚓激酶活峰值为397.6 U/mL。利用10 L发酵罐进行放大培养,经优化确定最佳菌体接种浓度为9±0.5 g/L,此条件下高密度发酵使前期培养时间缩短了11.5 h,蚓激酶活力达到1 098.2 U/mL,具有十分可观的工业应用潜力。

本文引用格式

江鹏 , 汤斌. . 蚓激酶基因在毕赤酵母中的表达及其发酵条件优化[J]. 食品与发酵工业, 2018 , 44(10) : 79 -83 . DOI: 10.13995/j.cnki.11-1802/ts.017178

Abstract

The encoding gene lks2 of lumbrokinase was cloned from Eisenia foetida and successfully expressed in Pichia pastoris GS115 with an activity of 254 U/mL. Optimization of culture conditions in shake flask showed that the best induction conditions were as follows: initial OD600=1.5, pH 5.5, temperature 28 ℃. Under these conditions, the activity of lumbrokinase peaked 397.6 U/mL at 84 h. Moreover, the amplification culture of recombinant P. pastoris GS115-pPIC9K-lks2 was occurred in a 10 L fermenter, of which the optimized inoculation concentration was 9±0.5 g/L. Under this high-density fermentation condition, the pre-culture time was shortened by 11.5 h and the activity of lumbrokinase was raised to 1 098.2 U/mL, which has a considerable potential for industrial application. In this study, the yield of lumbrokinase from P. pastoris was effectively increased by the optimization of induction conditions and the high-density fermentation, laying a foundation for the industrial production and application of lumbrokinase.
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