caa a22 4500 605499438 CHVBK 20210128100551.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00253-015-6816-6 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6816-6 Combining rational metabolic engineering and flux optimization strategies for efficient production of fumaric acid [Elektronische Daten] [Chan Song, Sang Lee] Fumaric acid is an important C4-dicarboxylic acid widely used in chemical, food, and pharmaceutical industries. Rational metabolic engineering together with flux optimization were performed for the development of an Escherichia coli strain capable of efficiently producing fumaric acid. The initial engineered strain, CWF4N overexpressing phosphoenolpyruvate carboxylase (PPC), produced 5.30g/L of fumaric acid. Optimization of PPC flux by examining 24 types of synthetic PPC expression vectors further increased the titer up to 5.72g/L with a yield of 0.432g/g·glucose. Overexpression of the succinate dehydrogenase complex (sdhCDAB) led to an increase in carbon yield up to 0.493g/g·glucose. Based on this mutant strain, citrate synthase (CS) was combinatorially overexpressed and balanced with PPC using 48 types of synthetic expression vectors. As a result, 6.24g/L of fumaric acid was produced with a yield of 0.500g/g·glucose. Fed-batch culture of this final strain allowed production of 25.5g/L of fumaric acid with a yield of 0.366g/g·glucose. Deletion of the aspA gene encoding aspartase and supplementation of aspartic acid further increased the fumaric acid titer to 35.1g/L with a yield of 0.490g/g·glucose. Springer-Verlag Berlin Heidelberg, 2015 Fumaric acid nationallicence Escherichia coli nationallicence Flux optimization nationallicence Metabolic engineering nationallicence Song Chan Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 plus program), Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), 305-701, Daejeon, Republic of Korea aut Lee Sang Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 plus program), Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), 305-701, Daejeon, Republic of Korea aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/20(2015-10-01), 8455-8464 0175-7598 99:20<8455 2015 99 253 https://doi.org/10.1007/s00253-015-6816-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6816-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Song Chan Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 plus program), Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), 305-701, Daejeon, Republic of Korea aut NATIONALLICENCE 700 1- Lee Sang Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 plus program), Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), 305-701, Daejeon, Republic of Korea aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/20(2015-10-01), 8455-8464 0175-7598 99:20<8455 2015 99 253