title: Three alcohol dehydrogenase genes and one acetyl-CoA synthetase gene are responsible for ethanol utilization in Yarrowia lipolytica.
abstract: The non-conventional yeast Yarrowia lipolytica is able to utilize a wide range of different substrates like glucose, glycerol, ethanol, acetate, proteins and various hydrophobic molecules. Although most metabolic pathways for the utilization of these substrates have been clarified by now, it was not clear whether ethanol is oxidized by alcohol dehydrogenases or by an alternative oxidation system inside the cell. In order to detect the genes that are required for ethanol utilization in Y. lipolytica, eight alcohol dehydrogenase (ADH) genes and one alcohol oxidase gene (FAO1) have been identified and respective deletion strains were tested for their ability to metabolize ethanol. As a result of this, we found that the availability of ADH1, ADH2 or ADH3 is required for ethanol utilization in Y. lipolytica. A strain with deletions in all three genes is lacking the ability to utilize ethanol as sole carbon source. Although Adh2p showed by far the highest enzyme activity in an in vitro assay, the availability of any of the three genes was sufficient to enable a decent growth. In addition to ADH1, ADH2 and ADH3, an acetyl-CoA synthetase encoding gene (ACS1) was found to be essential for ethanol utilization. As Y. lipolytica is a non-fermenting yeast, it is neither able to grow under anaerobic conditions nor to produce ethanol. To investigate whether Y. lipolytica may produce ethanol, the key genes of alcoholic fermentation in S. cerevisiae, ScADH1 and ScPDC1, were overexpressed in an ADH and an ACS1 deletion strain. However, instead of producing ethanol, the respective strains regained the ability to use ethanol as single carbon source and were still not able to grow under anaerobic conditions.
authors.affiliation: Institute of Microbiology, TU Dresden, Zellescher Weg 20b, 01217 Dresden, Germany. Electronic address: Michael.Gatter1@TU-Dresden.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Gatter
authors.affiliation: Institute of Microbiology, TU Dresden, Zellescher Weg 20b, 01217 Dresden, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Ottlik
authors.affiliation: Institute of Microbiology, TU Dresden, Zellescher Weg 20b, 01217 Dresden, Germany.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Kövesi
authors.affiliation: Institute of Microbiology, TU Dresden, Zellescher Weg 20b, 01217 Dresden, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Bauer
authors.affiliation: Institute of Microbiology, TU Dresden, Zellescher Weg 20b, 01217 Dresden, Germany.
authors.forename: Falk
authors.initials: F
authors.lastname: Matthäus
authors.affiliation: Institute of Microbiology, TU Dresden, Zellescher Weg 20b, 01217 Dresden, Germany.
authors.forename: Gerold
authors.initials: G
authors.lastname: Barth
mesh_terms: D000106:Acetate-CoA Ligase / Q000235:genetics*; D000332:Aerobiosis; D000426:Alcohol Dehydrogenase / Q000145:classification / Q000235:genetics*; D000693:Anaerobiosis; D004271:DNA, Fungal; D004789:Enzyme Activation / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D000431:Ethanol / Q000378:metabolism*; D005285:Fermentation; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal / Q000235:genetics; D010084:Oxidation-Reduction; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D014170:Transformation, Genetic; D025062:Yarrowia / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
chemical_list: D004271:DNA, Fungal; D000431:Ethanol; D000426:Alcohol Dehydrogenase; D000106:Acetate-CoA Ligase
keywords: Acetyl-CoA oxidase; Alcohol dehydrogenase; Ethanol metabolism; Ethanol utilization; Yarrowia lipolytica