Production and optimization of recombinant human augmenter of liver regeneration by Pichia pastoris
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- 作者：Zhimin Li ; Xubin Ping ; Qin Ye ; Xiudong Huang ; Zhifang Cao
- 关键词：Pichia pastoris ; Human augmenter of liver regeneration ; Methanol feeding strategy ; High cell density fermentation
- 刊名：Enzyme and Microbial Technology
- 出版年：2010
- 出版时间：6 October 2010
- 年：2010
- 卷：47
- 期：5
- 页码：222-227
- 全文大小：316 K
文摘
In this study, the fermentation of recombinant Pichia pastoris, which expresses heterogeneous human augmenter of liver regeneration (rhALR), has been optimized in a 5-L fermentor. The optimal methanol feeding method to optimize rhALR production was identified. Three methods of methanol feeding, methanol-stat, DO-stat and exponential feeding, were investigated to identify the method that would optimize rhALR production. It was determined that the methanol-stat method resulted in increased rhALR production, compared to the other two methods, which limit the amount of methanol available. Additionally, the results showed that rhALR expression levels could be enhanced by maintaining the pH at 6 and lowering the temperature from 30 to 25 °C. Supplementation with riboflavin during the induction phase also increased rhALR production. After these optimizations, rhALR was produced at a concentration of 424 mg/L, a three-fold improvement over original process.