caa a22 4500 605504245 CHVBK 20210128100615.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00253-014-6266-6 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6266-6 Exploiting mixtures of H2, CO2, and O2 for improved production of methacrylate precursor 2-hydroxyisobutyric acid by engineered Cupriavidus necator strains [Elektronische Daten] [Denise Przybylski, Thore Rohwerder, Cornelia Dilßner, Thomas Maskow, Hauke Harms, Roland Müller] Current manufacturing of most bulk chemicals through petrochemical routes considerably contributes to common concerns over the depletion of fossil carbon sources and greenhouse gas emissions. Sustainable future production of commodities thus requires the shift to renewable feedstocks in combination with established or newly developed synthesis routes. In this study, the potential of Cupriavidus necator H16 for autotrophic synthesis of the building block chemical 2-hydroxyisobutyric acid (2-HIBA) is evaluated. A novel biosynthetic pathway was implemented by heterologous expression of the 2-hydroxyisobutyryl-coenzyme A (2-HIB-CoA) mutase from Aquincola tertiaricarbonis L108, relying on a main intermediate of strain H16's C4 overflow metabolism, 3-hydroxybutyryl-CoA. The intention was to direct the latter to 2-HIBA instead or in addition to poly-3-hydroxybutyrate (PHB). Autotrophic growth and 2-HIBA (respectively, PHB) synthesis of wild-type and PHB-negative mutant strains were investigated producing maximum 2-HIBA titers of 3.2gL−1 and maximum specific 2-HIBA synthesis rates (q 2-HIBA) of about 16 and 175μmolg−1h−1, respectively. The obtained specific productivity was the highest reported to date for mutase-dependent 2-HIBA synthesis from heterotrophic and autotrophic substrates. Furthermore, expression of a G protein chaperone (MeaH) in addition to the 2-HIB-CoA mutase subunits yielded improved productivity. Analyzing the inhibition of growth and product synthesis due to substrate availability and product accumulation revealed a strong influence of 2-HIBA, when cells were cultivated at high titers. Nevertheless, the presented results imply that at the time the autotrophic synthesis route is superior to thus far established heterotrophic routes for production of 2-HIBA with C. necator. Springer-Verlag Berlin Heidelberg, 2014 2-Hydroxyisobutyric acid nationallicence Cupriavidus necator H16 PHB−4 nationallicence Building block nationallicence Autotrophic metabolism nationallicence Hydrogen nationallicence Carbon dioxide nationallicence Przybylski Denise Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut Rohwerder Thore Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut Dilßner Cornelia Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut Maskow Thomas Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut Harms Hauke Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut Müller Roland Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/5(2015-03-01), 2131-2145 0175-7598 99:5<2131 2015 99 253 https://doi.org/10.1007/s00253-014-6266-6 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/s00253-014-6266-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Przybylski Denise Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut NATIONALLICENCE 700 1- Rohwerder Thore Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut NATIONALLICENCE 700 1- Dilßner Cornelia Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut NATIONALLICENCE 700 1- Maskow Thomas Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut NATIONALLICENCE 700 1- Harms Hauke Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut NATIONALLICENCE 700 1- Müller Roland Department of Environmental Microbiology, Helmholtz Centre for Environmental Research UFZ, Permoserstrasse 15, 04318, Leipzig, Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/5(2015-03-01), 2131-2145 0175-7598 99:5<2131 2015 99 253