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通过代谢工程生产卡尔米尼酸

Dongsoo Yang^1,2,3, Woo Dae Jang^1,2, Sang Yup Lee^1,2,3,4

¹Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 plus program), Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Republic of Korea.

Journal of the American Chemical Society

|April 2, 2021

概括

研究人员利用细菌从葡萄糖中产生天然红色染料卡米尼酸. 大肠杆菌的这种生物合成途径为生产这种有价值的染料提供了传统昆虫养殖的可持续替代方案.

科学领域:

合成生物学
代谢工程
自然产品生物合成

背景情况:

传统上通过繁的昆虫养殖和净化生产的红色染料.
人们对开发微生物发酵方法来实现可持续的卡尔米尼酸生产非常感兴趣.

研究的目的:

在工程Escherichia coli中从葡萄糖中产生卡尔米尼酸的完整生物合成途径.
识别和优化卡尔米尼酸合成中的化和C-糖化关键酶.

主要方法:

来自Photorhabdus luminescens的II型聚化合成机制的优化.
用于生产黄酸的ZhuI和ZhuJ循环的同时表达.
生物化学查以鉴定甲基12-基酶 (DnrF) 和C-葡萄糖转移酶 (GtCGT) 的后续反应.
模拟和对接模拟以增强酶活性.
在大肠杆菌中进行代谢工程和食分批发酵.

主要成果:

工程化大肠杆菌成功产生了酸.
分别确定并验证了DnrF和GtCGT酶的基化和C- 葡萄糖化.
酶工程产生了2-5倍增强转化效率的突变物.
通过生产阿洛因,工程GtCGT证明了其广泛适用性.
从葡萄糖中获得0.63±0.02mg/L的卡尔米尼酸.

结论:

在工程化大肠杆菌中成功确定了卡尔米尼酸的完整生物合成途径.
这项研究表明一种可行的微生物发酵方法可以生产卡尔米尼酸.
使用的策略适用于设计其他工业相关的自然产品的生物合成途径.

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