title: Metabolic engineering of Escherichia coli for improving shikimate synthesis from glucose.
abstract: Shikimate is a key intermediate for the synthesis of the neuraminidase inhibitors. Microbial production of shikimate and related derivatives has the benefit of cost reduction when compared to traditional methods. In this study, an overproducing shikimate Escherichia coli strain was developed by rationally engineering certain metabolic pathways. To achieve this, the shikimate pathway was blocked by deletion of shikimate kinases and quinic acid/shikimate dehydrogenase. EIICB(glc) protein involved in the phosphotransferase system, and acetic acid pathway were also removed to increase the amount of available phosphoenolpyruvate and decrease byproduct formation, respectively. Thereafter, three critical enzymes of mutated 3-deoxy-D-arabinoheptulosonate-7-phosphate (DAHP) synthase (encoded by aroG(fbr)), PEP synthase (encoded by ppsA), and transketolase A (encoded by tktA) were modularly overexpressed and the resulting recombinant strain produced 1207 mg/L shikimate in shake flask cultures. Using the fed-batch process, 14.6g/L shikimate with a yield of 0.29 g/g glucose was generated in a 7-L bioreactor.
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China. Electronic address: xzchen@jiangnan.edu.cn.
authors.forename: Xianzhong
authors.initials: X
authors.lastname: Chen
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
authors.forename: Mingming
authors.initials: M
authors.lastname: Li
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Shen
authors.affiliation: Department of Biotechnology & Food Technology, Faculty of Applied Sciences, Durban University of Technology, P.O. Box 1334, Durban 4001, South Africa.
authors.forename: Govender
authors.initials: G
authors.lastname: Algasan
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
authors.forename: You
authors.initials: Y
authors.lastname: Fan
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology of Ministry of Education, Tianjin University of Science & Technology, Tianjin 300457, China.
authors.forename: Zhengxiang
authors.initials: Z
authors.lastname: Wang
mesh_terms: D010723:3-Deoxy-7-Phosphoheptulonate Synthase; D000429:Alcohol Oxidoreductases / Q000235:genetics; D019149:Bioreactors; D053898:Biosynthetic Pathways / Q000502:physiology*; D004926:Escherichia coli / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways / Q000502:physiology*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D017866:Phosphotransferases (Paired Acceptors); D011801:Quinic Acid / Q000378:metabolism; D012765:Shikimic Acid / Q000378:metabolism*; D014174:Transketolase
chemical_list: D011801:Quinic Acid; D012765:Shikimic Acid; D000429:Alcohol Oxidoreductases; C022641:Shikimate dehydrogenase; D014174:Transketolase; D010723:3-Deoxy-7-Phosphoheptulonate Synthase; D017853:Phosphotransferases (Alcohol Group Acceptor); C022642:shikimate kinase; D017866:Phosphotransferases (Paired Acceptors); C022509:pyruvate, water dikinase
keywords: Aromatic amino acid biosynthesis; Escherichia coli; Metabolic engineering; Shikimate