caa a22 4500 605504822 CHVBK 20210128100618.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s00253-015-6906-5 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6906-5 Production of carbon-13-labeled cadaverine by engineered Corynebacterium glutamicum using carbon-13-labeled methanol as co-substrate [Elektronische Daten] [Lennart Leßmeier, Johannes Pfeifenschneider, Marc Carnicer, Stephanie Heux, Jean-Charles Portais, Volker Wendisch] Methanol, a one-carbon compound, can be utilized by a variety of bacteria and other organisms as carbon and energy source and is regarded as a promising substrate for biotechnological production. In this study, a strain of non-methylotrophic Corynebacterium glutamicum, which was able to produce the polyamide building block cadaverine as non-native product, was engineered for co-utilization of methanol. Expression of the gene encoding NAD+-dependent methanol dehydrogenase (Mdh) from the natural methylotroph Bacillus methanolicus increased methanol oxidation. Deletion of the endogenous aldehyde dehydrogenase genes ald and fadH prevented methanol oxidation to carbon dioxide and formaldehyde detoxification via the linear formaldehyde dissimilation pathway. Heterologous expression of genes for the key enzymes hexulose-6-phosphate synthase and 6-phospho-3-hexuloisomerase of the ribulose monophosphate (RuMP) pathway in this strain restored growth in the presence of methanol or formaldehyde, which suggested efficient formaldehyde detoxification involving RuMP key enzymes. While growth with methanol as sole carbon source was not observed, the fate of 13C-methanol added as co-substrate to sugars was followed and the isotopologue distribution indicated incorporation into central metabolites and in vivo activity of the RuMP pathway. In addition, 13C-label from methanol was traced to the secreted product cadaverine. Thus, this synthetic biology approach led to a C. glutamicum strain that converted the non-natural carbon substrate methanol at least partially to the non-native product cadaverine. Springer-Verlag Berlin Heidelberg, 2015 Methylotrophy nationallicence Methanol nationallicence Corynebacterium glutamicum nationallicence Cadaverine nationallicence Diaminopentane nationallicence 13C-labeling nationallicence Bacillus methanolicus nationallicence Leßmeier Lennart Genetics of Prokaryotes, Faculty of Biology and Center for Biotechnology, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany aut Pfeifenschneider Johannes Genetics of Prokaryotes, Faculty of Biology and Center for Biotechnology, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany aut Carnicer Marc LISBP-INSA de Toulouse, 135 avenue de Rangueil, 31077, Toulouse CEDEX 04, France aut Heux Stephanie LISBP-INSA de Toulouse, 135 avenue de Rangueil, 31077, Toulouse CEDEX 04, France aut Portais Jean-Charles LISBP-INSA de Toulouse, 135 avenue de Rangueil, 31077, Toulouse CEDEX 04, France aut Wendisch Volker Genetics of Prokaryotes, Faculty of Biology and Center for Biotechnology, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 10163-10176 0175-7598 99:23<10163 2015 99 253 https://doi.org/10.1007/s00253-015-6906-5 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/s00253-015-6906-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Leßmeier Lennart Genetics of Prokaryotes, Faculty of Biology and Center for Biotechnology, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany aut NATIONALLICENCE 700 1- Pfeifenschneider Johannes Genetics of Prokaryotes, Faculty of Biology and Center for Biotechnology, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany aut NATIONALLICENCE 700 1- Carnicer Marc LISBP-INSA de Toulouse, 135 avenue de Rangueil, 31077, Toulouse CEDEX 04, France aut NATIONALLICENCE 700 1- Heux Stephanie LISBP-INSA de Toulouse, 135 avenue de Rangueil, 31077, Toulouse CEDEX 04, France aut NATIONALLICENCE 700 1- Portais Jean-Charles LISBP-INSA de Toulouse, 135 avenue de Rangueil, 31077, Toulouse CEDEX 04, France aut NATIONALLICENCE 700 1- Wendisch Volker Genetics of Prokaryotes, Faculty of Biology and Center for Biotechnology, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 10163-10176 0175-7598 99:23<10163 2015 99 253