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Methanol-Essential Growth of <i>Corynebacterium glutamicum</i>: Adaptive Laboratory Evolution Overcomes Limitation due to Methanethiol Assimilation Pathway
oleh: Guido Hennig, Carsten Haupka, Luciana F. Brito, Christian Rückert, Edern Cahoreau, Stéphanie Heux, Volker F. Wendisch
Format: | Article |
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Diterbitkan: | MDPI AG 2020-05-01 |
Deskripsi
Methanol is a sustainable substrate for biotechnology. In addition to natural methylotrophs, metabolic engineering has gained attention for transfer of methylotrophy. Here, we engineered <i>Corynebacterium glutamicum</i> for methanol-dependent growth with a sugar co-substrate. Heterologous expression of genes for methanol dehydrogenase from <i>Bacillus methanolicus</i> and of ribulose monophosphate pathway genes for hexulose phosphate synthase and isomerase from <i>Bacillus subtilis</i> 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 <i>C. glutamicum</i> to unleash the full potential of methanol as a carbon source in biotechnological processes.