High-density fermentation of <i>Escherichia coli</i> to express 4-hydroxyphenylacetate 3-hydroxylase and efficient biosynthesis of caffeic acid.

Hong Zhang; Jinlian Lin; Dinghang HU; Guiyou Liu; Lei Sun

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High-density fermentation of Escherichia coli to express 4-hydroxyphenylacetate 3-hydroxylase and efficient biosynthesis of caffeic acid.

Author: Hong ZHANG ¹ ; Jinlian LIN ¹ ; Dinghang HU ¹ ; Guiyou LIU ² ; Lei SUN ¹
Author Information

1. School of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong, China.
2. School of Life Sciences and Chemical Engineering, Jiangsu Second Normal University, Nanjing 211200, Jiangsu, China.
Publication Type:Journal Article
Keywords: 4-hydroxyphenylacetate 3-hydroxylase; caffeic acid; high-density fermentation; whole-cell biocatalysis
MeSH: Caffeic Acids; Escherichia coli/metabolism*; Fermentation; Metabolic Engineering; Mixed Function Oxygenases/metabolism*; Phenylacetates
From: Chinese Journal of Biotechnology 2022;38(9):3466-3477
CountryChina
Language:Chinese
Abstract: The 4-hydroxyphenylacetate 3-hydroxylase (4HPA3H), originated from Escherichia coli, converts p-coumaric acid to caffeic acid. In order to improve the efficiency of caffeic acid biosynthesis, we engineered E. coli for overexpression of 4HPA3H. The high-density fermentation of the engineered E. coli was conducted in a 5 L bioreactor. Subsequently, the conditions for whole-cell biocatalysis were optimized. The dry cell weight of the 4HPA3H-expressed strain reached 34.80 g/L. After incubated in the bioreactor for 6 h, 18.74 g/L (0.85 g/(L·OD₆₀₀)) of caffeic acid was obtained, with a conversion rate of 78.81% achieved. To the best of our knowledge, the titer of caffeic acid is the highest reported to date. The high-density fermentation of E. coli for overexpression of 4HPA3H and the efficient biosynthesis of caffeic acid may facilitate future large-scale production of caffeic acid.