ArticleBiotechnology for biofuels2021
Production of the biocommodities butanol and acetone from methanol with fluorescent FAST-tagged proteins using metabolically engineered strains of Eubacterium limosum.
Article in Biotechnology for biofuels, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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31 citing papers in PubMed, 60 citations in OpenAlex.
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- Deletion of atypical type II restriction genes in Clostridium cellulovorans using a Cas9-based gene editing system.Applied microbiology and biotechnology · 2025Article
- Recent advances in engineering non-native microorganisms for poly(3-hydroxybutyrate) production.World journal of microbiology & biotechnology · 2025Review
- Microbial single-cell applications under anoxic conditions.Applied and environmental microbiology · 2024Review
- Harnessing acetogenic bacteria for one-carbon valorization toward sustainable chemical production.RSC chemical biology · 2024Review
- Development of a Recombineering System for the AcetogenACS synthetic biology · 2024Article
- Single carbon metabolism - A new paradigm for microbial bioprocesses?Synthetic and systems biotechnology · 2024Review
- Constructing efficient bacterial cell factories to enable one-carbon utilization based on quantitative biology: A review.Quantitative biology (Beijing, China) · 2024Review
- Refining and illuminating acetogenic Eubacterium strains for reclassification and metabolic engineering.Microbial cell factories · 2024Article
- Engineered acetogenic bacteria as microbial cell factory for diversified biochemicals.Frontiers in bioengineering and biotechnology · 2024Review
- Green synthesis of (Frontiers in chemistry · 2024Article
- Microbiome imaging goes à la carte: Incorporating click chemistry into the fluorescence-activating and absorption-shifting tag (FAST) imaging platform.bioRxiv : the preprint server for biology · 2023Article
- Characterisation of acetogen formatotrophic potential using Eubacterium limosum.Applied microbiology and biotechnology · 2023Article
- Exploitation of a Type 1 Toxin-Antitoxin System as an Inducible Counter-Selective Marker for Genome Editing in the AcetogenMicroorganisms · 2023Article
- Application of the Fluorescence-Activating and Absorption-Shifting Tag (FAST) for Flow Cytometry in Methanogenic Archaea.Applied and environmental microbiology · 2023Article
- Heterologous 1,3-Propanediol Production Using Different RecombinantMicroorganisms · 2023Article
- Leveraging substrate flexibility and product selectivity of acetogens in two-stage systems for chemical production.Microbial biotechnology · 2023Review
- A fluorescent reporter system for anaerobic thermophiles.Frontiers in bioengineering and biotechnology · 2023Article
- Bacterial pathogens in pediatric appendicitis: a comprehensive retrospective study.Frontiers in cellular and infection microbiology · 2023Review
- Endogenous CRISPR/Cas systems for genome engineering in the acetogensFrontiers in bioengineering and biotechnology · 2023Article
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4 authors at 1 institution in 1 country.
Funding
Abstract
backgroundThe interest in using methanol as a substrate to cultivate acetogens increased in recent years since it can be sustainably produced from syngas and has the additional benefit of reducing greenhouse gas emissions. Eubacterium limosum is one of the few acetogens that can utilize methanol, is genetically accessible and, therefore, a promising candidate for the recombinant production of biocommodities from this C1 carbon source. Although several genetic tools are already available for certain acetogens including E. limosum, the use of brightly fluorescent reporter proteins is still limited.
resultsIn this study, we expanded the genetic toolbox of E. limosum by implementing the fluorescence-activating and absorption shifting tag (FAST) as a fluorescent reporter protein. Recombinant E. limosum strains that expressed the gene encoding FAST in an inducible and constitutive manner were constructed. Cultivation of these recombinant strains resulted in brightly fluorescent cells even under anaerobic conditions. Moreover, we produced the biocommodities butanol and acetone from methanol with recombinant E. limosum strains. Therefore, we used E. limosum cultures that produced FAST-tagged fusion proteins of the bifunctional acetaldehyde/alcohol dehydrogenase or the acetoacetate decarboxylase, respectively, and determined the fluorescence intensity and product concentrations during growth.
conclusionsThe addition of FAST as an oxygen-independent fluorescent reporter protein expands the genetic toolbox of E. limosum. Moreover, our results show that FAST-tagged fusion proteins can be constructed without negatively impacting the stability, functionality, and productivity of the resulting enzyme. Finally, butanol and acetone can be produced from methanol using recombinant E. limosum strains expressing genes encoding fluorescent FAST-tagged fusion proteins.
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