Metabolic engineering of Escherichia coli for the production of an antifouling agent zosteric acid

Peichao Zhang; Jing Gao; Haiyang Zhang; Yongzhen Wang; Zhen Liu; Sang Yup Lee; Xiangzhao Mao

doi:10.1016/j.ymben.2023.02.007

Metabolic engineering of Escherichia coli for the production of an antifouling agent zosteric acid

Metab Eng. 2023 Mar:76:247-259. doi: 10.1016/j.ymben.2023.02.007. Epub 2023 Feb 22.

Authors

Peichao Zhang¹, Jing Gao¹, Haiyang Zhang¹, Yongzhen Wang¹, Zhen Liu¹, Sang Yup Lee², Xiangzhao Mao³

Affiliations

¹ Qingdao Key Laboratory of Food Biotechnology, College of Food Science and Engineering, Ocean University of China, Qingdao, China.
² Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Four), BioProcess Engineering Research Center, Institute for the BioCentury, KAIST, Daejeon, Republic of Korea. Electronic address: leesy@kaist.ac.kr.
³ Qingdao Key Laboratory of Food Biotechnology, College of Food Science and Engineering, Ocean University of China, Qingdao, China; Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, Qingdao, China. Electronic address: xzhmao@ouc.edu.cn.

PMID: 36822462
DOI: 10.1016/j.ymben.2023.02.007

Abstract

Zosteric acid (ZA) is a Zostera species-derived, sulfated phenolic acid compound with antifouling activity and has gained much attention due to its nontoxic and biodegradable characteristics. However, the yield of Zostera species available for ZA extraction is limited by natural factors, such as season, latitude, light, and temperature. Here we report the development of metabolically engineered Escherichia coli strains capable of producing ZA from glucose and glycerol. First, intracellular availability of the sulfur donor 3'-phosphoadenosine-5'-phosphosulfate (PAPS) was enhanced by knocking out the cysH gene responsible for PAPS consumption and overexpressing the genes required for PAPS biosynthesis. Co-overexpression of the genes encoding tyrosine ammonia-lyase, sulfotransferase 1A1, ATP sulfurylase, and adenosine 5'-phosphosulfate kinase constructed ZA producing strain with enhanced PAPS supply. Second, the feedback-resistant forms of aroG and tyrA genes (encoding 3-deoxy-d-arabinoheptulosonate 7-phosphate synthase and chorismate mutase, respectively) were overexpressed to relieve the feedback regulation of L-tyrosine biosynthesis. Third, the pykA gene involved in phosphoenolpyruvate-consuming reaction, the regulator gene tyrR, the competing pathway gene pheA, and the ptsHIcrr genes essential for the PEP:carbohydrate phosphotransferase system were deleted. Moreover, all genes involved in the shikimate pathway and the talA, tktA, and tktB genes in the pentose phosphate pathway were examined for ZA production. The PTS-independent glucose uptake system, the expression vector system, and the carbon source were also optimized. As a result, the best-performing strain successfully produced 1.52 g L^-1 ZA and 1.30 g L^-1p-hydroxycinnamic acid from glucose and glycerol in a 700 mL fed-batch bioreactor.

Keywords: 3′-phosphoadenosine-5′-phosphosulfate; Escherichia coli; L-tyrosine; Metabolic engineering; Zosteric acid; p-Hydroxycinnamic acid.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biofouling* / prevention & control
Escherichia coli* / genetics
Escherichia coli* / metabolism
Glucose / genetics
Glucose / metabolism
Glycerol / metabolism
Metabolic Engineering

Substances

zosteric acid
Glycerol
shikimate
Glucose