title: Ethanol production by the hyperthermophilic archaeon Pyrococcus furiosus by expression of bacterial bifunctional alcohol dehydrogenases.
abstract: Ethanol is an important target for the renewable production of liquid transportation fuels. It can be produced biologically from pyruvate, via pyruvate decarboxylase, or from acetyl-CoA, by alcohol dehydrogenase E (AdhE). Thermophilic bacteria utilize AdhE, which is a bifunctional enzyme that contains both acetaldehyde dehydrogenase and alcohol dehydrogenase activities. Many of these organisms also contain a separate alcohol dehydrogenase (AdhA) that generates ethanol from acetaldehyde, although the role of AdhA in ethanol production is typically not clear. As acetyl-CoA is a key central metabolite that can be generated from a wide range of substrates, AdhE can serve as a single gene fuel module to produce ethanol through primary metabolic pathways. The focus here is on the hyperthermophilic archaeon Pyrococcus furiosus, which grows by fermenting sugar to acetate, CO2and H2. Previously, by the heterologous expression of adhA from a thermophilic bacterium, P. furiosus was shown to produce ethanol by a novel mechanism from acetate, mediated by AdhA and the native enzyme aldehyde oxidoreductase (AOR). In this study, the AOR gene was deleted from P. furiosus to evaluate ethanol production directly from acetyl-CoA by heterologous expression of the adhE gene from eight thermophilic bacteria. Only AdhEs from two Thermoanaerobacter strains showed significant activity in cell-free extracts of recombinant P. furiosus and supported ethanol production in vivo. In the AOR deletion background, the highest amount of ethanol (estimated 61% theoretical yield) was produced when adhE and adhA from Thermoanaerobacter were co-expressed.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA.
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Keller
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA.
authors.forename: Gina L
authors.initials: GL
authors.lastname: Lipscomb
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA.
authors.forename: Diep M
authors.initials: DM
authors.lastname: Nguyen
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA.
authors.forename: Alexander T
authors.initials: AT
authors.lastname: Crowley
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA.
authors.forename: Gerrit J
authors.initials: GJ
authors.lastname: Schut
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA.
authors.forename: Israel
authors.initials: I
authors.lastname: Scott
authors.affiliation: Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, 27695, USA.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Kelly
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA.
authors.forename: Michael W W
authors.initials: MWW
authors.lastname: Adams
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism*; D005285:Fermentation; D015870:Gene Expression; D060847:Metabolic Engineering; D020027:Pyrococcus furiosus / Q000235:genetics / Q000378:metabolism*; D046968:Thermoanaerobacter / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D000431:Ethanol; D000426:Alcohol Dehydrogenase