Transcriptome analysis of Pichia pastoris in response to ethanol stress

Peng Gao1 Jian Ding1 Xu Zhang1 Yue Zhao1 Meng Zhang1 Minjie Gao1 Jianrong Wu1 Xiaobei Zhan1

(1.Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China 214122)

【Abstract】Effective expression of pIFN-α in recombinant Pichia pastoris was conducted in a 5 L fermentor. Ethanol accumulation during the late glycerol feeding period inhibited heterologous protein expression. Comparative transcriptome analysis was thus performed to compare the gene transcription profiles of Pichia pastoris KM71H in high and low ethanol concentration environments. The results showed that during the glycerol cultivation stage, 545 genes (265 up-regulated and 280 down-regulated) were differentially expressed with ethanol stress. These genes were mainly involved in protein synthesis, energy metabolism, cell cycle and peroxisome metabolism. During the methanol induction stage, 294 genes (171 up-regulated and 123 down-regulated) were differentially expressed, which were mainly related to methanol metabolism, amino acid metabolism and protein synthesis. Ethanol stress increased protein misfolding and reduced structural integrity of ribosome and mitochondria during cultivation stage, and led to the failure of endoplasmic reticulum stress removal and damaged amino acid metabolism during induction stage in Pichia pastoris.

【Keywords】 Pichia pastoris; heterologous protein; ethanol stress; transcriptome;

【DOI】

【Funds】 National Natural Science Foundation of China (No. 31301408) National Natural Science Foundation of China(No.31301408) Natural Science Foundation of Jiangsu Province (No. BK20130122) Natural Science Foundation of Jiangsu Province(No.BK20130122) China Postdoctoral Science Foundation (No. 2014M551501) China Postdoctoral Science Foundation(No.2014M551501)

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(Translated by ZHAO Bo)

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This Article

ISSN:1000-3061

CN: 11-1998/Q

Vol 32, No. 05, Pages 584-598

May 2016

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Abstract

  • 1 Materials and methods
  • 2 Results and analysis
  • 3 Discussion
  • References