title: Production of protocatechuic acid by Corynebacterium glutamicum expressing chorismate-pyruvate lyase from Escherichia coli.
abstract: Protocatechuic acid (3,4-dihydroxybenzoic acid; PCA) serves as a building block for polymers and pharmaceuticals. In this study, the biosynthetic pathway for PCA from glucose was engineered in Corynebacterium glutamicum. The pathway to PCA-employed elements of the chorismate pathway by using chorismate-pyruvate lyase (CPL) and 4-hydroxybenzoate hydroxylase (4-HBA hydroxylase). As C. glutamicum has the potential to synthesize the aromatic amino acid intermediate chorismate and possesses 4-HBA hydroxylase, we focused on expressing Escherichia coli CPL in a phenylalanine-producing strain of C. glutamicum ATCC21420. To secrete PCA, the gene (ubiC) encoding CPL from E. coli was expressed in C. glutamicum ATCC 21420 (strain F(UbiC)). The formation of 28.8 mg/L of extracellular 4-HBA (36 h) and 213 ± 29 mg/L of extracellular PCA (80 h) was obtained by the C. glutamicum strain F(UbiC) from glucose. The strain ATCC21420 was also found to produce extracellular PCA. PCA fermentation was performed using C. glutamicum strain F(UbiC) in a bioreactor at the optimized pH of 7.5. C. glutamicum F(UbiC) produced 615 ± 2.1 mg/L of PCA from 50 g/L of glucose after 72 h. Further, fed-batch fermentation of PCA by C. glutamicum F(UbiC) was performed with feedings of glucose every 24 h. The maximum production of PCA (1140.0 ± 11.6 mg/L) was achieved when 117.0 g/L of glucose was added over 96 h of fed-batch fermentation.
authors.affiliation: Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Okai
authors.affiliation: Technology Alliance Development Department, New Business Development Business Unit, Teijin Limited, Kasumigaseki Common Gate West Tower 2-1, 3 Kasumigaseki, Chiyoda-ku, Tokyo, 100-8585, Japan.
authors.forename: Takanori
authors.initials: T
authors.lastname: Miyoshi
authors.affiliation: Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan.
authors.forename: Yasunobu
authors.initials: Y
authors.lastname: Takeshima
authors.affiliation: Corporate Strategic Technology Department, Technology Center, Teijin Limited, Kasumigaseki Common Gate West Tower 2-1, 3 Kasumigaseki, Chiyoda-ku, Tokyo, 100-8585, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kuwahara
authors.affiliation: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Ogino
authors.affiliation: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan. akondo@kobe-u.ac.jp.|Biomass Engineering Program, RIKEN, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan. akondo@kobe-u.ac.jp.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kondo
mesh_terms: D019149:Bioreactors; D048230:Corynebacterium glutamicum / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D062385:Hydroxybenzoates / Q000378:metabolism*; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D007652:Oxo-Acid-Lyases / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D062385:Hydroxybenzoates; D011994:Recombinant Proteins; C009091:protocatechuic acid; D007652:Oxo-Acid-Lyases; C075960:chorismate pyruvate lyase
keywords: Chorismate-pyruvate lyase; Corynebacterium glutamicum; Fed-batch fermentation; Protocatechuic acid