Engineering the central biosynthetic and secondary metabolic pathways of Pseudomonas aeruginosa strain PA1201 to improve phenazine-1-carboxylic acid production
Engineering the central biosynthetic and secondary metabolic pathways of Pseudomonas aeruginosa strain PA1201 to improve phenazine-1-carboxylic acid production
复制标题
对铜绿假单胞菌 PA1201 菌株的中央生物合成和次级代谢途径进行工程设计，以提高吩嗪-1-羧酸的产量
DOI：
10.1016/j.ymben.2015.09.003
复制
发表时间：
2015-11-01
影响因子：
8.4
通讯作者：
He, Ya-Wen
中科院分区：
文献类型：
--
作者：
Jin, Kaiming;Zhou, Lian;He, Ya-Wen
次级代谢物phenazine-1-carboxylic acid （PCA）是新注册生物农药Shenqinmycin的重要成分。我们采用基因工程、启动子工程和蛋白质工程相结合的方法，对产生pca的铜绿假单胞菌PA1201的中心生物合成和次级代谢途径进行了修饰。通过阻断主成分转化，增强主成分外排泵送，使菌株PA-IV的主成分产率提高54.6倍；(2)通过过量生产两种DAHP合成酶和阻断21种次级代谢物的合成，增加了PCA生物合成途径的代谢通量；(3)通过改造5种利用choris酸的酶，增加PCA前体的供应；(4)对两个PCA生物合成基因簇的启动子进行工程化。菌株PA-IV在补料间歇发酵中产生了9882 mg/L的PCA，是目前工业菌株的两倍。菌株PA-IV遗传稳定，在细胞毒性方面与大肠杆菌相当。(C) 2015年国际代谢工程学会。Elsevier Inc.出版。版权所有。
The secondary metabolite phenazine-1-carboxylic acid (PCA) is an important component of the newly registered biopesticide Shenqinmycin. We used a combined method involving gene, promoter, and protein engineering to modify the central biosynthetic and secondary metabolic pathways in the PCA-producing Pseudomonas aeruginosa strain PA1201. The PCA yield of the resulting strain PA-IV was increased 54.6-fold via the following strategies: (1) blocking PCA conversion and enhancing PCA efflux pumping; (2) increasing metabolic flux towards the PCA biosynthetic pathway through the overproduction of two DAHP synthases and blocking the synthesis of 21 secondary metabolites; (3) increasing the PCA precursor supply through the engineering of five chorismate-utilizing enzymes; (4) engineering the promoters of two PCA biosynthetic gene clusters. Strain PA-IV produced 9882 mg/L PCA in fed-batch fermenation, which is twice as much as that produced by the current industrial strain. Strain PA-IV was also genetically stable and comparable to Escherichia coli in cytotoxicity. (C) 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.