Muts型基因重组巴斯德毕赤酵母高密度发酵过程中甲醇流加的控制

在采用Muts 表型基因重组巴斯德毕赤酵母 (Pichiapastoris)发酵生产血管生长抑制因子(angiostatin)的过程中 ,诱导表达阶段甲醇的质量浓度对血管生长抑制因子的表达至关重要 .因重组Muts 型巴斯德毕赤酵母利用甲醇极慢 ,按文献方法流加甲醇时 ,甲醇逐渐积累达 30 g/L ,血管生长抑制素表达水平仅为 4mg/L .采用甲醇检测与控制系统对甲醇流加进行反馈控制后 ,甲醇质量浓度可稳定控制在设定范围 .采用此系统控制诱导阶段的甲醇流加 ,同时手动流加适量甘油以促进细胞生长 ,血管生长抑制因子表达水平量最高可达 89mg/L .     

补料分批培养耐乙酸大肠杆菌生产人表皮生长因子

An acetate-tolerant mutant of Escherichia coli DH5α, DA19, was used for secretory production of human epidermal growth factor （hEGF） whose expression was under the control of phoA promoter. The recombinant cells were cultured in a chemically defined medium, and glucose was added at different specific provision rates during the growth and expression phases. It was found that pH had a significant effect on the extracellular hEGF production. The extracellular hEGF concentration was 75.5mg·L^-1, 5.2-fold of the level reached at pH 7.0, even though more acetate was produced. Nitrogen source was limited in the later growth phase. Supplementation of ammonium promoted the consumption of phosphate and reduced the time to exhaust phosphate, but the extracellular hEGF production was similar to that without supplementation of ammonium.