构建基因工程大肠杆菌对L-Threo-3-Hydroxyaspartic酸的De Novo生产进行基因工程
Jing Guo1,2,3, Jiayi Cui4, Mingyue Xun4
1CAS Key Laboratory of Bio-Based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, 266101, China. guojing@qibebt.ac.cn.
这项研究设计了大肠杆菌从葡萄糖中产生新的L-threo-3-hydroxyaspartic acid (L-THA). 工程菌株的最终度为2.87g/L,证明了工业L-THA生物合成的可行发酵平台.
科学领域:
- 生物技术是生物技术.
- 代谢工程是代谢工程.
- 合成生物学 合成生物学
背景情况:
- L-threo-3-hydroxyaspartic acid (L-THA) 是一种具有显著生物活性的非蛋白质性氨基酸.
- 目前的L-THA合成方法依赖于昂贵的基质,如L-酸或L-氨酸,这阻碍了大规模应用.
研究的目的:
- 将大肠杆菌 (E. coli) 设计为细胞工厂,通过发酵直接从葡萄糖中合成L-THA.
- 为优化工程化大肠杆菌菌株以提高L-THA生产标位.
主要方法:
- 在E. coli中编码阿斯巴拉金氧酶的Streptomyces coelicolor asnO基因的异质表达.
- 过度表达原生大肠杆菌基因aspC (阿斯巴达胺转移酶) 和asnB (阿斯巴拉金合成酶) 以改善L-THA前体供应.
- 代谢工程策略包括aspA (阿斯巴达酶) 的过度表达和lysC (阿斯巴达酸激酶III) 的淘汰,以增强L-阿斯巴达酸池.
主要成果:
- 这种工程化的大肠杆菌菌株成功地从葡萄糖中产生L-THA.
- 优化工作导致L-THA标位增加了5.6倍,在摇瓶发酵中达到278.3mg/L.
- 料批发发酵在32小时内达到2.87g/L的最终L-THA标位.
结论:
- 大肠杆菌发酵是从葡萄糖中新生合成L-THA的可行平台.
- 开发的代谢工程策略显著提高了L-THA的生产.
- 这种方法对L-THA生产的工业规模应用具有前景.
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