枯草芽孢杆菌产角蛋白酶的发酵过程优化

Optimization of Culture Process Conditions and Feeding Strategy for Keratinase by Recombinant Bacillus subtilis

DOI：10.3969/j.issn.1673-1689.2014.05.003

中文关键词: 重组枯草芽孢杆菌 流加策略 发酵优化 角蛋白酶

英文关键词: recombinant Bacillus subtilis, feeding strategy, fermentation optimization, keratinase

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中文摘要:

为了实现重组角蛋白酶在基因工程菌Bacillus subtilis WB600中的高效生产，在3 L发酵罐中对发酵条件和补料策略进行了优化，建立了补料分批发酵工艺。研究发现，两阶段控制pH，前6小时控制pH 7.0，后期控制pH 8.0、DO 20%，发酵过程以葡萄糖为流加碳源，7~11 h以μ为0.15 h-1指数流加、12~15 h以μ为0.1 h-1指数流加、16~27 h恒速流加葡萄糖3 g/（L·h），在30 L发酵罐水平上，27 h酶活达到864 U/mL，生产强度为31.8 U/（mL·h），国外报道的酶活达到2 900 mL，酶活在国内属领先水平，为重组枯草芽孢杆菌产角蛋白酶的工业化生产打下了坚实的基础。

英文摘要:

In order to achieve the high production of keratinase by recombinant Bacillus subtilis WB600, fermentation conditions and feeding strategy were optimized in 3 L fermentor, and a fed-batch fermentation was establishde. The results showed that, using a two-stage pH control strategy in which pH was controlled at 7.0 for the first 4 h and then 8.0 for remaining time and a mixed feeding strategy in which keeping the exponential feeding from 6 h to 15 h(6~11 h μ was 0.15 h-1, 11~15 h μ was 0.1 h-1) and then constant-fed fermentation for remaining time, the keratinase activity reached 864 U/mL. The keratinase activity reached a leading level at home, and the keratinase reached 2 900 U/mL overseas.This work reached a high keratinase productivity and it is a stable theroret-ics for the fermention sacle-up process of keratinase.

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