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- Title
Methanol-Essential Growth of Corynebacterium glutamicum: Adaptive Laboratory Evolution Overcomes Limitation due to Methanethiol Assimilation Pathway.
- Authors
Hennig, Guido; Haupka, Carsten; Brito, Luciana F.; Rückert, Christian; Cahoreau, Edern; Heux, Stéphanie; Wendisch, Volker F.
- Abstract
Methanol is a sustainable substrate for biotechnology. In addition to natural methylotrophs, metabolic engineering has gained attention for transfer of methylotrophy. Here, we engineered Corynebacterium glutamicum for methanol-dependent growth with a sugar co-substrate. Heterologous expression of genes for methanol dehydrogenase from Bacillus methanolicus and of ribulose monophosphate pathway genes for hexulose phosphate synthase and isomerase from Bacillus subtilis enabled methanol-dependent growth of mutants carrying one of two independent metabolic cut-offs, i.e., either lacking ribose-5-phosphate isomerase or ribulose-5-phosphate epimerase. Whole genome sequencing of strains selected by adaptive laboratory evolution (ALE) for faster methanol-dependent growth was performed. Subsequently, three mutations were identified that caused improved methanol-dependent growth by (1) increased plasmid copy numbers, (2) enhanced riboflavin supply and (3) reduced formation of the methionine-analogue O-methyl-homoserine in the methanethiol pathway. Our findings serve as a foundation for the engineering of C. glutamicum to unleash the full potential of methanol as a carbon source in biotechnological processes.
- Subjects
CORYNEBACTERIUM glutamicum; METHANETHIOL; BACILLUS subtilis; METHIONINE; NUCLEOTIDE sequencing; ISOMERASES
- Publication
International Journal of Molecular Sciences, 2020, Vol 21, Issue 10, p3617
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms21103617