Production of the biocommodities butanol and acetone from methanol with fluorescent FAST-tagged proteins using metabolically engineered strains of Eubacterium limosum

Abstract Background The interest in using methanol as a substrate to cultivate acetogens increased recent years since it can be sustainably produced from syngas and has the additional benefit of reducing greenhouse gas emissions. Eubacterium limosum is one few that utilize methanol, genetically accessible and, therefore, promising candidate for recombinant production biocommodities this C1 carbon source. Although several genetic tools are already available certain including E. limosum, use brightly fluorescent reporter proteins still limited. Results In study, we expanded toolbox by implementing fluorescence-activating absorption shifting tag (FAST) protein. Recombinant strains expressed gene encoding FAST an inducible constitutive manner were constructed. Cultivation these resulted cells even under anaerobic conditions. Moreover, butanol acetone with strains. Therefore, used cultures FAST-tagged fusion bifunctional acetaldehyde/alcohol dehydrogenase or acetoacetate decarboxylase, respectively, determined fluorescence intensity product concentrations during growth. Conclusions addition oxygen-independent protein expands . our results show constructed without negatively impacting stability, functionality, productivity resulting enzyme. Finally, expressing genes proteins.