恶臭假单胞菌
鼠李糖脂
合成生物学
化学
异源的
生物化学
生物
食品科学
基因
计算生物学
铜绿假单胞菌
细菌
遗传学
作者
Aiping Pang,Yun Wang,Teng Zhang,Feng Gao,Jidong Shen,Lianggang Huang,Junping Zhou,Bo Zhang,Zhi‐Qiang Liu,Yu‐Guo Zheng
标识
DOI:10.1016/j.biortech.2023.130220
摘要
Pseudomonas putida KT2440, a GRAS strain, has been used for synthesizing bulk and fine chemicals. However, the gene editing tool to metabolically engineer KT2440 showed low efficiency. In this study, a novel sacB-based system pK51mobsacB was established to improve the efficiency for marker-free gene disruption. Then the rhamnolipid synthetic pathway was introduced in KT2440 and genes of the competitive pathways were deleted to lower the metabolic burden based on pK51mobsacB. A series of endogenous and synthetic promoters were used for fine tuning rhlAB expression. The limited supply of dTDP-L-rhamnose was enhanced by heterologous rmlBDAC expression. Cell growth and rhamnolipid production were well balanced by using glucose/glycerol as mixed carbon sources. The final strain produced 3.64 g/L at shake-flask and 19.77 g/L rhamnolipid in a 5 L fermenter, the highest obtained among metabolically engineered KT2440, which implied the potential of KT2440 as a promising microbial cell factory for industrial rhamnolipid production.
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