在重组大肠杆菌BL21中进行5 - 氨基诺列维林酸的工程生产
Zhiwen Zhu1, Bing Fu1,2, Jiajie Lu1
1College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, People's Republic of China.
Preparative biochemistry & biotechnology
|November 5, 2024
概括
改造了大肠杆菌BL21以促进5氨基氨基酸 (ALA) 的产生. 优化代谢途径和基因表达实现了创纪录的ALA 2953 mg/L,为工业应用铺平了道路.
科学领域:
- 代谢工程是代谢工程.
- 合成生物学 合成生物学
- 生物技术是生物技术.
背景情况:
- 5氨基联氨酸 (ALA) 是血红素和叶绿素合成的关键前体,在医学和农业中具有应用.
- 目前的ALA生产方法在产量和效率方面存在局限性,需要代谢工程方法.
- 大肠杆菌为微生物生产像ALA这样有价值的生物化学物质提供了一个强大的平台.
研究的目的:
- 改造大肠杆菌BL21中的C5通路,以提高5-氨基氨基酸 (ALA) 的产生.
- 为了优化对ALA生物合成的代谢流量,并最大限度地减少其下游转化.
- 建立一个代谢工程的大肠杆菌菌株,以实现高效和可扩展的ALA工业生产.
主要方法:
- 过度表达ALA合成酶基因 (gltX,hemA,hemL) 以增加ALA前体供应.
- 过度表达sRNA RyhB以降低ALA脱水酶的调节,防止ALA的转化.
- 通过CRISPR-Cas9调解,对arcA进行淘汰,以缓解反抑制和增强呼吸.
- 通过响应表面方法来优化发酵条件.
主要成果:
- 改造后的大肠杆菌BL21显著提高了ALA的产生.
- 这种工程菌株在1升摇瓶中获得了2953毫克/升的最大ALA产量.
- 代谢修饰成功地将细胞资源重定向到ALA积累.
结论:
- 代谢工程策略,包括基因过度表达和淘汰,对于促进大肠杆菌中ALA的产生是有效的.
- 开发的工程菌株具有成本效益高的工业规模ALA制造的潜力.
- 这项研究为使用微生物平台进一步优化和扩大ALA生物合成提供了基础.
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