title: Biosynthesis of higher alcohol flavour compounds by the yeast Saccharomyces cerevisiae: impact of oxygen availability and responses to glucose pulse in minimal growth medium with leucine as sole nitrogen source.
abstract: Higher alcohol formation by yeast is of great interest in the field of fermented beverages. Among them, medium-chain alcohols impact greatly the final flavour profile of alcoholic beverages, even at low concentrations. It is widely accepted that amino acid metabolism in yeasts directly influences higher alcohol formation, especially the catabolism of aromatic and branched-chain amino acids. However, it is not clear how the availability of oxygen and glucose metabolism influence the final higher alcohol levels in fermented beverages. Here, using an industrial Brazilian cachaça strain of Saccharomyces cerevisiae, we investigated the effect of oxygen limitation and glucose pulse on the accumulation of higher alcohol compounds in batch cultures, with glucose (20 g/l) and leucine (9.8 g/l) as the carbon and nitrogen sources, respectively. Fermentative metabolites and CO2 /O2 balance were analysed in order to correlate the results with physiological data. Our results show that the accumulation of isoamyl alcohol by yeast is independent of oxygen availability in the medium, depending mainly on leucine, α-keto-acids and/or NADH pools. High-availability leucine experiments showed a novel and unexpected accumulation of isobutanol, active amyl alcohol and 2-phenylethanol, which could be attributed to de novo biosynthesis of valine, isoleucine and phenylalanine and subsequent outflow of these pathways. In carbon-exhausted conditions, our results also describe, for the first time, the metabolization of isoamyl alcohol, isobutanol, active amyl alcohol but not of 2-phenylethanol, by yeast strains in stationary phase, suggesting a role for these higher alcohols as carbon source for cell maintenance and/or redox homeostasis during this physiological phase.
authors.affiliation: Centre for Strategic Technologies of the Northeast (CETENE), Pernambuco, Brazil; Interdepartmental Research Group in Metabolic Engineering, Federal University of Pernambuco, Brazil.
authors.forename: Esteban
authors.initials: E
authors.lastname: Espinosa Vidal
authors.forename: Marcos Antonio
authors.initials: MA
authors.lastname: de Morais
authors.forename: Jean Marie
authors.initials: JM
authors.lastname: François
authors.forename: Gustavo M
authors.initials: GM
authors.lastname: de Billerbeck
mesh_terms: D000434:Alcoholic Beverages / Q000032:analysis / Q000382:microbiology*; D000438:Alcohols / Q000032:analysis / Q000378:metabolism*; D001938:Brazil; D003470:Culture Media / Q000378:metabolism*; D005285:Fermentation; D005421:Flavoring Agents / Q000032:analysis / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D007930:Leucine / Q000378:metabolism*; D009584:Nitrogen / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000438:Alcohols; D003470:Culture Media; D005421:Flavoring Agents; D029701:Saccharomyces cerevisiae Proteins; D007930:Leucine; D005947:Glucose; D009584:Nitrogen; D010100:Oxygen
keywords: Ehrlich pathway; alcoholic beverage; amino acid metabolism; flavour compounds; industrial alcoholic fermentation