Engineering of 2,3-Butanediol Dehydrogenase To Reduce Acetoin Formation by Glycerol-Overproducing, Low-AlcoholSaccharomyces cerevisiae
| Autor: | Sylvie Dequin, María Rosario Fernández, Josep A. Biosca, Anne Julien, Maryam Ehsani |
|---|---|
| Přispěvatelé: | Sciences Pour l'Oenologie (SPO), Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Université Montpellier 1 (UM1)-Université de Montpellier (UM)-Institut National de la Recherche Agronomique (INRA), Lallemand S.A.S., Universitat Autònoma de Barcelona (UAB) |
| Rok vydání: | 2009 |
| Předmět: |
Glycerol
[SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT] Gene Expression Dehydrogenase Diacetyl Saccharomyces cerevisiae Biology Models Biological Applied Microbiology and Biotechnology 03 medical and health sciences chemistry.chemical_compound Butylene Glycols 030304 developmental biology Fermentation in winemaking 0303 health sciences Ethanol Ecology 030306 microbiology Acetoin Yeast Alcohol Oxidoreductases Yeast in winemaking chemistry Biochemistry Fermentation Food Microbiology Genetic Engineering Metabolic Networks and Pathways Food Science Biotechnology |
| Zdroj: | Applied and Environmental Microbiology Applied and Environmental Microbiology, American Society for Microbiology, 2009, 75 (10), pp.3196-3205. ⟨10.1128/aem.02157-08⟩ |
| ISSN: | 1098-5336 0099-2240 |
| DOI: | 10.1128/aem.02157-08 |
| Popis: | Appl. Environ. Microbiol. ISI Document Delivery No.: 443KL Times Cited: 14 Cited Reference Count: 45 Ehsani, Maryam Fernandez, Maria R. Biosca, Josep A. Julien, Anne Dequin, Sylvie Ministry of Education and Science [BMC-2003-09606] This work was supported in part by a grant from the Ministry of Education and Science (BMC-2003-09606), Spain. Amer soc microbiology Washington; International audience; Engineered Saccharomyces cerevisiae strains overexpressing GPD1, which codes for glycerol-3-phosphate dehydrogenase, and lacking the acetaldehyde dehydrogenase Ald6 display large-scale diversion of the carbon flux from ethanol toward glycerol without accumulating acetate. Although GPD1 ald6 strains have great potential for reducing the ethanol contents in wines, one major side effect is the accumulation of acetoin, having a negative sensory impact on wine. Acetoin is reduced to 2,3-butanediol by the NADH-dependent 2,3-butanediol dehydrogenase Bdh1. In order to investigate the influence of potential factors limiting this reaction, we overexpressed BDH1, coding for native NADH-dependent Bdh1, and the engineered gene BDH1 221,222,223, coding for an NADPH-dependent Bdh1 enzyme with the amino acid changes 221 EIA 223 to 221 SRS 223, in a glycerol-overproducing wine yeast. We have shown that both the amount of Bdh1 and the NADH availability limit the 2,3-butanediol dehydrogenase reaction. During wine fermentation, however, the major limiting factor was the level of synthesis of Bdh1. Consistent with this finding, the overproduction of native or engineered Bdh1 made it possible to redirect 85 to 90% of the accumulated acetoin into 2,3-butanediol, a compound with neutral sensory characteristics. In addition, the production of diacetyl, a compound causing off-flavor in alcoholic beverages, whose production is increased in glycerol-overproducing yeast cells, was decreased by half. The production of higher alcohols and esters, which was slightly decreased or unchanged in GPD1 ald6 cells compared to that in the control cells, was not further modified in BDH1 cells. Overall, rerouting carbons toward glycerol and 2,3-butanediol represents a new milestone in the engineering of a low-alcohol yeast with desirable organoleptic features, permitting the decrease of the ethanol contents in wines by up to 3 degrees. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|