Strain engineering with FAST is getting a hot topic in the anaerobe community these days. A team led by Dr. Frank Bengelsdorf of Ulm University has just added Acetobacterium woodii to the continuously expanding list of FAST-tagged anaerobes. They indeed just published in Appl. Microbiol. Biotechnol. 2022 in collaboration with the University of Nottingham.
Happy and proud to welcome the Synthetic Biology Research Centre (SBRC) of the University of Nottingham in The Twinkle Factory community!
Luc Lenglet, C.E.O. Twinkle Bioscience
This work is part of the rush towards a more circular economy. Namely, it is about generating value-added products from gaseous waste streams. For this purpose, biological processes, i.e., fermentation, are ahead of the race. Acetogenic bacteria can indeed convert industrial waste gas streams containing CO2 to various organic acids and alcohols. One promising candidate for waste streams valorization is lactate. Lactate and lactic acid have indeed a wide range of applications, e.g., in food, cosmetics, pharmaceuticals, or as a precursor for poly-lactic acid (PLA), a biodegradable plastic. Moreover, lactate is a promising feedstock in co-cultures to produce value-added products such as propionic acid, butyrate and caproate.
The paper discloses how FAST helped to construct Acetobacterium woodii strains. The objective was the autotrophic lactate production using H2 + CO2 as energy and carbon source. One key roadblock at engineering anaerobes is indeed the lack of fluorescent reporters. Yet FAST does not require any oxygen nor maturation delay. As a result, FAST allows instant fluorescence reporting even in fully anoxic conditions. Here, FAST-tagged D-lactate dehydrogenase (LDHD) originating from Leuconostoc mesenteroides was expressed in A. woodii. Cells showed a clear and bright fluorescence during exponential growth, as well as in the stationary phase. The authors concluded that FAST provides a new reporter tool to quickly analyze gene expression over growth experiments of A. woodii.
The Twinkle Factory provides commercial fluorogens to users of FAST and splitFAST in the research community.
More reading on strain engineering with FAST
- Appl. Microbiol. Biotechnol. 2022) – Autotrophic lactate production from H2 + CO2 using recombinant and fluorescent FAST-tagged Acetobacterium woodii strains
- ACS Synth. Biol. 2022 – Establishment of Green- and Red-Fluorescent Reporter Proteins Based on the Fluorescence-Activating and Absorption-Shifting Tag for Use in Acetogenic and Solventogenic Anaerobes
- Nat. Commun. 2021 – Auxin-mediated protein depletion for metabolic engineering in terpene-producing yeast
- Biotechnol. Biofuels 2021 – Production of the Biocommodities Butanol and Acetone from Methanol with Fluorescent FAST-tagged Proteins using Metabolically Engineered Strains of Eubacterium Limosum
- mBio 2020 – Interspecies Microbial Fusion and Large-Scale Exchange of Cytoplasmic Proteins and RNA in a Syntrophic Clostridium Coculture
- Appl. Environ. Microbiol. 2019 – A Strongly Fluorescing Anaerobic Reporter and Protein-Tagging System for Clostridium Organisms Based on the Fluorescence-Activating and Absorption-Shifting Tag Protein (FAST)