Du Jin, Zhang Aijun, Hao Jian'an, Wang Jing
The Institute of Seawater Desalination and Multipurpose Utilization, SOA, Tianjin, 300192, China.
Biotechnol Lett. 2017 Jul;39(7):1041-1048. doi: 10.1007/s10529-017-2333-2. Epub 2017 Apr 3.
To engineer Escherichia coli for the heterologous production of di-rhamnolipids, which are important biosurfactants but mainly produced by opportunistic pathogen Pseudomonas aeruginosa.
The codon-optimized rhlAB and rhlC genes originating from P. aeruginosa and Burkholderia pseudomallei were combinatorially expressed in E. coli to produce di-rhamnolipids with varied congeners compositions. Genes involved in endogenous upstream pathways (rhamnose and fatty acids synthesis) were co-overexpressed with rhlAB-rhlC, resulting in variations of rhamnolipids production and congeners compositions. Under the shake-flask condition, co-overexpression of rfbD with rhlAB-rhlC increased rhamnolipids production (0.64 ± 0.02 g l) than that in strain only expressing rhlAB-rhlC (0.446 ± 0.009 g l), which was mainly composed of di-rhamnolipids congeners Rha-Rha-C-C.
Biosynthesis of di-rhamnolipids and variations of congeners composition in genetically engineered E. coli strains were achieved via combiniations of mono-/di-rhamnolipids synthesis modules and endogenous upstream modules.
构建能够异源生产二鼠李糖脂的大肠杆菌,二鼠李糖脂是重要的生物表面活性剂,但主要由机会致病菌铜绿假单胞菌产生。
源自铜绿假单胞菌和类鼻疽伯克霍尔德菌的密码子优化的rhlAB和rhlC基因在大肠杆菌中组合表达,以产生具有不同同系物组成的二鼠李糖脂。与rhlAB - rhlC共同过表达参与内源性上游途径(鼠李糖和脂肪酸合成)的基因,导致鼠李糖脂产量和同系物组成发生变化。在摇瓶条件下,rfbD与rhlAB - rhlC共同过表达比仅表达rhlAB - rhlC的菌株(0.446±0.009 g/l)的鼠李糖脂产量增加(0.64±0.02 g/l),其主要由二鼠李糖脂同系物Rha - Rha - C - C组成。
通过单/二鼠李糖脂合成模块与内源性上游模块的组合,实现了基因工程大肠杆菌菌株中二鼠李糖脂的生物合成及其同系物组成的变化。