Metabolic engineering of Caldicellulosiruptor bescii for 2,3-butanediol production from unpretreated lignocellulosic biomass and metabolic strategies for improving yields and titers.

Autor: Tanwee TNN; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA., Lipscomb GL; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA., Vailionis JL; Department of Cell and Molecular Biology, College of the Environment and Life Sciences, University of Rhode Island, Kingston, Rhode Island, USA., Zhang K; Department of Cell and Molecular Biology, College of the Environment and Life Sciences, University of Rhode Island, Kingston, Rhode Island, USA., Bing RG; Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, North Carolina, USA., O'Quinn HC; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA., Poole FL; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA., Zhang Y; Department of Cell and Molecular Biology, College of the Environment and Life Sciences, University of Rhode Island, Kingston, Rhode Island, USA., Kelly RM; Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, North Carolina, USA., Adams MWW; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA.
Jazyk: angličtina
Zdroj: Applied and environmental microbiology [Appl Environ Microbiol] 2024 Jan 24; Vol. 90 (1), pp. e0195123. Date of Electronic Publication: 2023 Dec 22.
DOI: 10.1128/aem.01951-23
Abstrakt: The platform chemical 2,3-butanediol (2,3-BDO) is used to derive products, such as 1,3-butadiene and methyl ethyl ketone, for the chemical and fuel production industries. Efficient microbial 2,3-BDO production at industrial scales has not been achieved yet for various reasons, including product inhibition to host organisms, mixed stereospecificity in product formation, and dependence on expensive substrates (i.e., glucose). In this study, we explore engineering of a 2,3-BDO pathway in Caldicellulosiruptor bescii , an extremely thermophilic (optimal growth temperature = 78°C) and anaerobic bacterium that can break down crystalline cellulose and hemicellulose into fermentable C 5 and C 6 sugars. In addition , C. bescii grows on unpretreated plant biomass, such as switchgrass. Biosynthesis of 2,3-BDO involves three steps: two molecules of pyruvate are condensed into acetolactate; acetolactate is decarboxylated to acetoin, and finally, acetoin is reduced to 2,3-BDO. C. bescii natively produces acetoin; therefore, in order to complete the 2,3-BDO biosynthetic pathway, C. bescii was engineered to produce a secondary alcohol dehydrogenase (sADH) to catalyze the final step. Two previously characterized, thermostable sADH enzymes with high affinity for acetoin, one from a bacterium and one from an archaeon, were tested independently. When either sADH was present in C. bescii, the recombinant strains were able to produce up to 2.5-mM 2,3-BDO from crystalline cellulose and xylan and 0.2-mM 2,3-BDO directly from unpretreated switchgrass. This serves as the basis for higher yields and productivities, and to this end, limiting factors and potential genetic targets for further optimization were assessed using the genome-scale metabolic model of C. bescii .IMPORTANCELignocellulosic plant biomass as the substrate for microbial synthesis of 2,3-butanediol is one of the major keys toward cost-effective bio-based production of this chemical at an industrial scale. However, deconstruction of biomass to release the sugars for microbial growth currently requires expensive thermochemical and enzymatic pretreatments. In this study, the thermo-cellulolytic bacterium Caldicellulosiruptor bescii was successfully engineered to produce 2,3-butanediol from cellulose, xylan, and directly from unpretreated switchgrass. Genome-scale metabolic modeling of C. bescii was applied to adjust carbon and redox fluxes to maximize productivity of 2,3-butanediol, thereby revealing bottlenecks that require genetic modifications.
Competing Interests: The authors declare no conflict of interest.
Databáze: MEDLINE
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