重组大肠杆菌bl(puc-hyp)产羟脯氨酸的补料分批培养.doc


重组大肠杆菌 BL21(pUC19-Hyp)产羟脯氨
酸的补料分批培养#
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摘要:利用本实验室构建的组成型重组大肠杆菌 BL21(pUC19-Hyp)为出发菌株,在 7 L 发酵
罐中,运用间歇流加、指数流加、恒速流加三种流加碳源的方式进行补料分批培养。结果表
明:以 g/min 恒速流加为最优,在发酵开始 44 h 时羟脯氨酸(Hydroxyproline,Hyp)的浓
度达到最高,浓度为 g/L,脯氨酸(Proline,Pro)转化率为 81%,此时细胞干重为
g/L,残糖浓度为 g/L。Hyp 含量与摇瓶发酵时的 g/L 相比提高了大约 30 倍,比日
本株式会社的发酵产量提高了 g/L[1],发酵过程中消耗的葡萄糖与产生的 Hyp 质量之比
约为 4:1。发酵液中的氨基酸分析结果表明,除 Pro、Hyp 外的其他氨基酸含量均低于 1 mg/L,
发酵液中主要氨基酸为 Pro 和 Hyp。
关键词:发酵工程;重组大肠杆菌;羟脯氨酸(Hyp);恒速流加
中图分类号:Q819
Production of Hydroxyproline by fed-batch culture of novel
binant Escherichia coli BL21(pUC19-Hyp)
Yuan chunwei1,2, Zhang shegnli1,2, Liu hedong1,2, He yanchun1,2, Zhang zhenyu1,2
(1. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University,
JiangSu WuXi 214122;
2. School of Biotechnology, Jiangnan University, JiangSu WuXi 214122)
Abstract: We processed the supplementary fe-batch cultivation in the 7 L fermentor by use of the
constitutive binant escherichia coli BL21 (pUC19 - Hyp), starting strain, constructed in
our lab and three different ways of flow and carbon source which contains intermittent flow
addition, index flow addition and constant speed flow. The results showed that the optimal method
was constant speed flow with g/min. After 44 hours of fermentation, the concentration of
Hydroxyproline was the highest which was g/L; Proline conversion rate is 81%; the cell dry
weight is g/L; residual sugar concentration is g/L. Compared with shaking flask
fermentation with concentration of g/L, Hyp content improved about 30 times and at the
same time, the fermentation yield improved g/L than Japan Ltd [1]. In the process of
fermentation, the ratio of glucose consumption and product of Hyp is about 4:1. The analysis of
amino acids in the fermentation liquid showed that, Pro and Hyp are the main amino acids in
fermentation liquid, and except for Pro and Hyp, the other amino aci