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Biosynthesis of 2,4-diacetylphloroglucinol from glucose using engineered Escherichia coli
Wen Liu1, Rubing Zhang2, Mo Xian3
1CAS Key Laboratory of Bio-Based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, 266101, China.
Abstract:
In order to produce 2,4-diacetylphloroglucinol (2,4-DAPG) in E. coli, the key synthases coding by phlACBD gene cluster from the strain Pseudomonas fluorescens CHA0 were overexpressed in E. coli BL21 (DE3). The marA, phlE and acc genes were also overexpressed to enhance 2,4-DAPG biosynthesis. Then the fermentation conditions were optimized to improve the concentration of 2,4-DAPG. The results showed that the recombinant E. coli could produce few 2,4-DAPG with only the phlACBD gene cluster. The synthetic ability of 2,4-DAPG could be increased by expressing the acc, marA and phlE genes in shake-flasks cultivation. The effects of phloroglucinol, initial pH, temperature and trace elements on 2,4-DAPG biosynthesis were also investigated. Based on the optimal fermentation conditions obtained from the shake-flasks cultivation, fed-batch fermentation of strain Z3 in a 5 L bioreactor was conducted to produce 2,4-DAPG. Finally, the concentration of 2,4-DAPG was 179 mg/L after induction for 36 h by fed-batch fermentation. To the best of our knowledge, this is the highest 2,4-DAPG production reported in E. coli. This work showed the potential application of engineered E. coli to get high production of target compounds.
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