期刊
METABOLIC ENGINEERING
卷 45, 期 -, 页码 134-141出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2017.12.006
关键词
Phenolic compounds; Pyrogallol; Shikimate pathway; Metabolic engineering; Escherichia coli
资金
- National Natural Science Foundation of China [21376017, 21406010, 21636001]
- Program of Introducing Talents of Discipline to Universities (111 project) [B13005]
- Program for Changjiang Scholars and Innovative Research Team in Universities in China [IRT13045]
- National High Technology Research and Development Program of China (863 Program) [2015AA021001]
Pyrogallol is a simple phenolic compound that serves as an attractive chemical with broad applications in food, agricultural, dyeing, printing, cosmetic, photography, chemical and pharmaceutical industries owing to its antioxidant, antibacterial, antiseptic, anticancer, oxygen-absorbing and strong reducing properties. Currently, pyrogallol is commercially produced by thermal decarboxylation of gallic acid under high temperature and pressure. However, this process is limited by the inaccessible raw material, the strict reaction conditions and the relatively low product yield. Here, we report establishment of a novel and efficient biosynthetic pathway for the synthesis of pyrogallol. First, we identified and characterized an efficient 2,3-dihydroxybenzoic acid (2,3-DHBA) 1-monoxygenase from a series of oxygenases and hydroxylases based on the structural similarity in the substrates and products, which enabled non-natural production of pyrogallol from 2,3-DHBA. Then, over-expression of 2,3-DHBA synthase and 2,3-DHBA 1-monoxygenase achieved synthesis of pyrogallol in Escherichia coli, with a titer of 201.52 mg/L at 24 h. Further optimizations by enhancement of the carbon flux through the shikimate pathway, modular optimization of the pathway and alleviation of the pyrogallol autoxidation boosted pyrogallol titer to 1035.75 mg/L in shake flask experiments. This work constructed an efficient microbial platform for gram per liter level production of pyrogallol, indicating the great potential for industrial biomanufacturing of pyrogallol.
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