caa a22 4500 605498008 CHVBK 20210128100544.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-6092-x doi (NATIONALLICENCE)springer-10.1007/s00253-014-6092-x Metabolic engineering of itaconate production in Escherichia coli [Elektronische Daten] [Kiira Vuoristo, Astrid Mars, Jose Sangra, Jan Springer, Gerrit Eggink, Johan Sanders, Ruud Weusthuis] Interest in sustainable development has led to efforts to replace petrochemical-based monomers with biomass-based ones. Itaconic acid, a C5-dicarboxylic acid, is a potential monomer for the chemical industry with many prospective applications. cis-aconitate decarboxylase (CadA) is the key enzyme of itaconate production, converting the citric acid cycle intermediate cis-aconitate into itaconate. Heterologous expression of cadA from Aspergillus terreus in Escherichia coli resulted in low CadA activities and production of trace amounts of itaconate on Luria-Bertani (LB) medium (<10mg/L). CadA was primarily present as inclusion bodies, explaining the low activity. The activity was significantly improved by using lower cultivation temperatures and mineral medium, and this resulted in enhanced itaconate titres (240mg/L). The itaconate titre was further increased by introducing citrate synthase and aconitase from Corynebacterium glutamicum and by deleting the genes encoding phosphate acetyltransferase and lactate dehydrogenase. These deletions in E. coli's central metabolism resulted in the accumulation of pyruvate, which is a precursor for itaconate biosynthesis. As a result, itaconate production in aerobic bioreactor cultures was increased up to 690mg/L. The maximum yield obtained was 0.09mol itaconate/mol glucose. Strategies for a further improvement of itaconate production are discussed. Springer-Verlag Berlin Heidelberg, 2014 Itaconic acid nationallicence Escherichia coli nationallicence Metabolic engineering nationallicence Citrate synthase nationallicence Aconitase nationallicence cis -aconitate decarboxylase nationallicence Vuoristo Kiira Bioprocess Engineering, Wageningen University, Wageningen, The Netherlands aut Mars Astrid Biobased Products, Wageningen University and Research Centre, Wageningen UR PO Box 8129, 6700 EV, Wageningen, The Netherlands aut Sangra Jose Bioprocess Engineering, Wageningen University, Wageningen, The Netherlands aut Springer Jan Biobased Products, Wageningen University and Research Centre, Wageningen UR PO Box 8129, 6700 EV, Wageningen, The Netherlands aut Eggink Gerrit Biobased Products, Wageningen University and Research Centre, Wageningen UR PO Box 8129, 6700 EV, Wageningen, The Netherlands aut Sanders Johan Biobased Products, Wageningen University and Research Centre, Wageningen UR PO Box 8129, 6700 EV, Wageningen, The Netherlands aut Weusthuis Ruud Bioprocess Engineering, Wageningen University, Wageningen, The Netherlands aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 221-228 0175-7598 99:1<221 2015 99 253 https://doi.org/10.1007/s00253-014-6092-x 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/s00253-014-6092-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Vuoristo Kiira Bioprocess Engineering, Wageningen University, Wageningen, The Netherlands aut NATIONALLICENCE 700 1- Mars Astrid Biobased Products, Wageningen University and Research Centre, Wageningen UR PO Box 8129, 6700 EV, Wageningen, The Netherlands aut NATIONALLICENCE 700 1- Sangra Jose Bioprocess Engineering, Wageningen University, Wageningen, The Netherlands aut NATIONALLICENCE 700 1- Springer Jan Biobased Products, Wageningen University and Research Centre, Wageningen UR PO Box 8129, 6700 EV, Wageningen, The Netherlands aut NATIONALLICENCE 700 1- Eggink Gerrit Biobased Products, Wageningen University and Research Centre, Wageningen UR PO Box 8129, 6700 EV, Wageningen, The Netherlands aut NATIONALLICENCE 700 1- Sanders Johan Biobased Products, Wageningen University and Research Centre, Wageningen UR PO Box 8129, 6700 EV, Wageningen, The Netherlands aut NATIONALLICENCE 700 1- Weusthuis Ruud Bioprocess Engineering, Wageningen University, Wageningen, The Netherlands aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 221-228 0175-7598 99:1<221 2015 99 253