caa a22 4500 605500436 CHVBK 20210128100556.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s00253-015-6497-1 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6497-1 Construction of a novel phenol synthetic pathway in Escherichia coli through 4-hydroxybenzoate decarboxylation [Elektronische Daten] [Liangtian Miao, Qingyan Li, Aipo Diao, Xueli Zhang, Yanhe Ma] Phenol is a bulk chemical with lots of applications in the chemical industry. Fermentative production of phenol had been realized in both Pseudomonas putida and Escherichia coli by recruiting tyrosine phenol-lyase (TPL). The TPL pathway needs tyrosine as the direct precursor for phenol production. In this work, a novel phenol synthetic pathway was created in E. coli by recruiting 4-hydroxybenzoate decarboxylase, which can convert 4-hydroxybenzoate to phenol and carbon dioxide. Activating 3-deoxy-d-arabino-heptulosonate-7-phosphate (DAHP) synthase and chorismate pyruvate lyase (UbiC) through plasmid overexpression led to 7- and 69-fold increase of phenol production, respectively, demonstrating that these two enzymes were the rate-limiting steps for phenol production. Genetically stable strains were then obtained by gene integration and gene modulation directly in chromosome. Phenol titer increased 147-fold (from 1.7 to 250mg/L) after modulating the DAHP synthase, UbiC, and 4-hydroxybenzoate decarboxylase genes in chromosome. Five solvents were tested for two-phase extractive fermentation to eliminate phenol toxicity to E. coli cells. Tributyrin and dibutyl phthalate were the best two solvents for improving phenol production, leading to 23 and 30% increase of total phenol production, respectively. Two-phase fed-batch fermentation of the best strain Phe009 was performed in a 7L fermentor, which produced 9.51g/L phenol with a yield of 0.061g/g glucose. Springer-Verlag Berlin Heidelberg, 2015 Phenol nationallicence 4-hydroxybenzoate decarboxylase nationallicence Two-phase extractive fermentation nationallicence Escherichia coli nationallicence Miao Liangtian Tianjin University of Science & Technology, 300457, Tianjin, China aut Li Qingyan Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 300308, Tianjin, China aut Diao Aipo Tianjin University of Science & Technology, 300457, Tianjin, China aut Zhang Xueli Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 300308, Tianjin, China aut Ma Yanhe Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 300308, Tianjin, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/12(2015-06-01), 5163-5173 0175-7598 99:12<5163 2015 99 253 https://doi.org/10.1007/s00253-015-6497-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6497-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Miao Liangtian Tianjin University of Science & Technology, 300457, Tianjin, China aut NATIONALLICENCE 700 1- Li Qingyan Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 300308, Tianjin, China aut NATIONALLICENCE 700 1- Diao Aipo Tianjin University of Science & Technology, 300457, Tianjin, China aut NATIONALLICENCE 700 1- Zhang Xueli Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 300308, Tianjin, China aut NATIONALLICENCE 700 1- Ma Yanhe Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 300308, Tianjin, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/12(2015-06-01), 5163-5173 0175-7598 99:12<5163 2015 99 253