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产L-丝氨酸谷氨酸棒杆菌的代谢工程改造和代谢流分析

, PP. 295-303

Keywords: 谷氨酸棒杆菌,L-丝氨酸,胞内代谢物,代谢工程,基元模型分析

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Abstract:

作为胞内一碳单位的主要来源,L-丝氨酸对维持菌体生理代谢具有不可替代的作用,导致其难于实现以糖质为原料的直接发酵法生产.本研究采用系统代谢工程策略,构建了基因工程菌CorynebacteriumglutamicumSER-8.连续补料发酵实验显示,SER-8的比生长速率降低,L-丝氨酸的积累量在不同的生长时期呈现变化.代谢流分析证明,过表达3-磷酸甘油酸激酶使糖酵解途径和三羧酸循环增强,过表达脱敏型3-磷酸甘油酸脱氢酶加强了L-丝氨酸合成途径.敲除激活蛋白GlyR部分减弱了L-丝氨酸向甘氨酸的转化,代谢流比率仍有38.2%.实验结果表明,胞内L-丝氨酸的代谢对维持C.glutamicum的生长至关重要,而通过引入GCV裂解系统,分解过量的甘氨酸,同时增加胞内一碳单位的供应将是未来L-丝氨酸代谢工程改造的新靶点.

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