title: Increased incorporation of gaseous CO2into succinate by Escherichia coli overexpressing carbonic anhydrase and phosphoenolpyruvate carboxylase genes.
abstract: Carbon dioxide (CO2) is an abundant and cheap carbon source that is partly responsible for global warming in the atmosphere. The objective of this study was to construct a recombinant E. coli strain that can show enhanced production of succinate derived from CO2. In this study, we confirmed the enhancement of utilization by analyzing succinate containing one carbon-13 (13C) derived from13CO2. Firstly, the carbonic anhydrase gene (SP(-)HCCA) derived from Hahella chejuensis KCTC 2396 was over-expressed to enhance carbon flux toward bicarbonate ion (HCO3-) synthesis in E. coli. The phosphoenolpyruvate carboxylase gene (ppc) was over-expressed to enhance the production of oxaloacetate by enhancing the carbon flux. Compared with the control strain, the percentage of the succinate containing one13C (succinate119) to total succinate was enhanced by approximately 2.80-fold and the amount of succinate119also increased by approximately 4.09-fold in SGJS120. Secondly, the lactate dehydrogenase gene (ldhA) was deleted to re-direct the utilization of the carbon source from glucose to enhance succinate production in SGJS120. However, ldhA deletion did not increase CO2utilization in SJGS120. Finally, the phosphotransferase system gene (ptsG) and pyruvate kinase F gene (pykF) were deleted to increase the amount of phosphoenolpyruvate (PEP). SGJS126 (pykF deletion strain) showed the highest increase, which was 6.05-fold higher than the control strain. From the results, SP(-)HCCA overexpression and pykF deletion may be useful for enhancing CO2utilization in E. coli. Additionally, engineered strains showed the potential to reduce the cost of succinate production by using an industrially cheaper carbon source such as CO2and converting CO2to a valuable chemical.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Sogang University, Seoul 121-742, Republic of Korea.
authors.forename: Soohyun
authors.initials: S
authors.lastname: Park
authors.affiliation: Department of Chemistry, Sogang University, Seoul, 121-742, Republic of Korea.
authors.forename: Jae-Ung
authors.initials: JU
authors.lastname: Lee
authors.affiliation: C1 Gas Refinery R&D Center, Sogang University, Seoul, 121-742,Republic of Korea.
authors.forename: Sukhyeong
authors.initials: S
authors.lastname: Cho
authors.affiliation: Department of Chemical and Biomolecular Engineering, Sogang University, Seoul 121-742, Republic of Korea.
authors.forename: Hyeonsoo
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Sogang University, Seoul, 121-742, Republic of Korea.
authors.forename: Han Bin
authors.initials: HB
authors.lastname: Oh
authors.affiliation: Department of Biotechnology and Bioinformatics, Korea University, Sejong, Chungnam 339-700, Republic of Korea. Electronic address: spack@korea.ac.kr.
authors.forename: Seung Pil
authors.initials: SP
authors.lastname: Pack
authors.affiliation: Department of Chemical and Biomolecular Engineering, Sogang University, Seoul 121-742, Republic of Korea; C1 Gas Refinery R&D Center, Sogang University, Seoul, 121-742,Republic of Korea. Electronic address: jinwonlee@sogang.ac.kr.
authors.forename: Jinwon
authors.initials: J
authors.lastname: Lee
mesh_terms: D002245:Carbon Dioxide / Q000378:metabolism*; D002256:Carbonic Anhydrases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways; D010730:Phosphoenolpyruvate Carboxylase / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D019802:Succinic Acid / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D002245:Carbon Dioxide; D019802:Succinic Acid; D010730:Phosphoenolpyruvate Carboxylase; D002256:Carbonic Anhydrases
keywords: Carbon dioxide; Carbonic anhydrase; Escherichia coli; Phosphoenolpyruvate carboxylase; Succinate