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Manipulating the levels of ethyl acetate and isoamyl acetate formation during the production of wine and brandy

Thesis (MSc)--University of Stellenbosch, 2002.

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Main Author: Bayly, Jennifer Carr,1977-
Other Authors: Du Toit, M.
Format: Thesis
Language:en_ZA
Published: Stellenbosch : Stellenbosch University 2012
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access_status_str Open Access
author Bayly, Jennifer Carr,1977-
author2 Du Toit, M.
author_browse Bayly, Jennifer Carr,1977-
Du Toit, M.
author_facet Du Toit, M.
Bayly, Jennifer Carr,1977-
author_sort Bayly, Jennifer Carr,1977-
collection Thesis
dc_rights_str_mv Stellenbosch University
description Thesis (MSc)--University of Stellenbosch, 2002.
format Thesis
id oai:scholar.sun.ac.za:10019.1/52926
institution Stellenbosch University (South Africa)
language en_ZA
last_indexed 2026-06-10T12:43:35.721Z
license_str Other — see source repository
provenance_str_mv Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository
publishDate 2012
publishDateRange 2012
publishDateSort 2012
publisher Stellenbosch : Stellenbosch University
publisherStr Stellenbosch : Stellenbosch University
record_format dspace
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spelling oai:scholar.sun.ac.za:10019.1/52926 Manipulating the levels of ethyl acetate and isoamyl acetate formation during the production of wine and brandy Bayly, Jennifer Carr,1977- Du Toit, M. Pretorius, I. S. Lambrechts, M. G. Stellenbosch University. Faculty of AgriSciences. Dept. of Viticulture and Oenology. Institute for Wine Biotechnology. Brandy Wine and wine making Ethyl acetoacetate Dissertations -- Wine biotechnology Theses -- Wine biotechnology Thesis (MSc)--University of Stellenbosch, 2002. ENGLISH ABSTRACT: The production of wine is a complex process, which involves the conversion of sugar in grape must to ethanol, carbon dioxide and other byproducts. The principal organism in winemaking is yeast, of which Saccharomyces cerevisiae is the most important due to its ability to survive winemaking conditions, its GRAS (Generally Regarded As Safe) status and the favourable flavours it imparts during the winemaking process. However, due to the demands of the consumer and the emergence of sophisticated wine markets, a demand is developing for specialised yeast strains with enhanced and new oenological properties. For these reasons, research into the contribution of wine yeast to the aroma bouquet as well the influence of wine or brandy maturation in wood on the aroma bouquet is important for consumer demands to be met. The fruity aroma of wine is associated with esters, which are produced during the alcoholic fermentation by yeast. Important acetate esters in wine and brandy are ethyl acetate, which has a fruity, solvent-like aroma, and isoamyl acetate, which has a banana-like aroma. These esters are produced through the action of acetyltransferases (AATases), which catalyse the reaction between a higher alcohol and acyl Coenzyme A. Esters are mainly a product of alcoholic fermentation. However, their concentration changes during wood maturation and it has been found that the concentration of acetate esters can increase during the maturation period. In this study, the aim was to investigate the influence of AATase I and AATase II, which are encoded by the ATF1 and ATF2 genes respectively, on the aroma bouquet of wine and brandy. Therefore, the first objective of this study was to clone the ATF2 gene from a commercial wine yeast strain and to overexpress this gene in a commercial wine yeast strain and in a wine yeast strain that already has the A TF1 gene overexpressed. Disruption cassettes were also designed in order to disrupt the ATF1 and ATF2 genes in a commercial wine yeast strain. The resultant recombinant wine yeast strains were used for the production of wine and brandy. GC analyses and tasting trials were conducted to determine the effect of the overexpression or disruption of these genes on the aroma bouquet of wine. The results obtained indicated that there are differences in the aroma bouquet of wine and brandy when changes are made in gene expression. The results indicated that the A TF1 gene plays a large role in the production of ethyl and isoamyl acetate. When this gene was overexpressed, the level of ethyl acetate was 5.6-fold more than that of the control and the level of isoamyl acetate was 3.5-fold higher than that of the control. However, no increase in ethyl acetate or isoamyl acetate was observed when the A TF2 gene was overexpressed. An increase in 2-phenylethyl acetate and diethyl succinate was observed in brandy, although there was a decrease in total ester concentration. A decrease in acetic acid was also observed in the brandy produced, which could be an indication of ester production. Similarly, no increase in ethyl acetate or isoamyl acetate was observed in the wine or brandy produced when both the ATF1 and ATF2 genes were overexpressed in a single yeast. Once again, a marked decrease was observed in acetic acid concentration in both the wine and brandy. In conclusion, it is clear that changes in gene expression can change the aroma profile of wine or brandy. However, the role of the ATF2 gene still remains unclear and further studies are needed to clarify its role in yeast. Future studies involving the effect of wood maturation on ester concentration will also be of importance, so that the winemaker or distiller can make a product that suits the ever-changing market. AFRIKAANSE OPSOMMING: Die produksie van wyn is 'n komplekse proses wat die omskakeling van die suiker in mos tot etanol, koolstofdioksied en ander byprodukte tot gevolg het. Die hooforganisme betrokke in die wynmaakproses is gis, waarvan Saccharomyces cerevisiae as een van die belangrikste geag word as gevolg van die vermoë daarvan om onder die wynfermentasietoestande te kan oorleef, die "GRAS"-status (Generally Regarded As Safe) daarvan en die invloed wat dit op die aroma van die uiteindelike produk het weens die werking daarvan gedurende alkoholiese fermentasie. Die behoefte aan wyn met nuwe, verbeterde eienskappe het die vraag na meer gespesialiseerde gisrasse deur beide die verbruiker en nuwe wynmarkte gedurende die afgelope paar jaar drasties laat toeneem. Dit is om dié redes dat navorsing oor die bydrae van wyngis en houtveroudering tot die aroma van beide wyn en brandewyn so belangrik geag word. Die vrugtige aroma van wyn word geassosieer met die esters wat gedurende die alkoholiese fermentasie deur gis gevorm word. Die belangrikste asetaatesters in wyn en brandewyn is etielasetaat, wat vir 'n oplosmiddelagtige, vrugtige aroma bekend is, en isoamielasetaat, wat 'n piesangaroma veroorsaak. Die esters word geproduseer deur die werking van asetieltransferases (AATases), wat as katalis in die reaksie tussen 'n hoër alkohol en asetiel-Ko-ensiem A optree. Alhoewel esters hoofsaaklik 'n produk van alkoholiese fermentasie is, wissel die konsentrasie daarvan gedurende houtveroudering. Daar is gevind dat die konsentrasie van die asetaatesters gedurende die verouderingsproses kan verhoog. Die studie het ten doelom die invloed van AATase I en AATase II, wat onderskeidelik deur die ATF1- en ATF2-gene geënkodeer word, op die aroma van wyn en brandewyn te ondersoek. Die eerste doelwit van die studie was vervolgens om die ATF2-geen vanaf 'n kommersiële wyngisras te kloneer en dit daarna te ooruitdruk in 'n kommersiële wyngisras, asook die geen te ooruitdruk in 'n kommersiële wyngisras wat reeds die ATF1-geen ooruitdruk. Disrupsiekassette is ook vir die disrupsie van die ATF1- en ATF2-gene in 'n kommersiële wyngisras ontwerp. Die rekombinante wyngisrasse wat gedurnde die studie gemaak is, is vir die produksie van wyn en brandewyn gebruik. Gas chromatografise-ontledings en sensoriese evaluerings is ook op die wyn en brandewyn uitgevoer. Die resultate van die studie het bewys dat daar wel veranderings plaasvind wanneer 'n verandering in geenuitdrukking gemaak is. Die resultate het weereens bevestig dat die ATF1-geen 'n belangrike rol in die produksie van etiel- en isoamielasetaat speel. Wanneer die ATF1-geen ooruitgedruk is, is die etielasetaatproduksie 5.6 keer meer en die isoamielasetaatproduksie 3.5 keer meer as in die kontrole. Die ooruitdrukking van die ATF2-geen het geen verhoging in etielasetaat of isoamielasetaat of in totale esters in die wyn getoon nie, alhoewel die ras 2.7 keer meer diëtielsuksinaat geproduseer het. In die brandewyn wat geproduseer is met die gisras waarin ATF2 ooruitgedruk is, was daar wel 'n verlaging in die asynsuur, wat 'n aanduiding van estervorming kan wees, alhoewel die totale esters wat geproduseer is minder was as in die kontrole. 'n Verhoging in diëtielsuksinaat en 2-fenielasetaat is ook gevind. Daar is geen verhoging in etiel- of isoamielasetaat getoon wanneer die ATF1- en ATF2-geen saam ooruitgedruk is nie. Die ras het minder totale sure in wyn en brandewyn geproduseer en ook geen verhoging in totale esters getoon nie. Uit die resultate is dit duidelik dat veranderings in geenuitdrukking 'n verandering in die aromaprofiel van wyn en brandewyn kan veroorsaak. Die rol van dié A TF2-geen is nog steeds onduidelik en verdere studies sal moet plaasvind om die rol van die geen te verduidelik. Studies wat konsentreer op die invloed van houtveroudering op esterkonsentrasie is ook belangrik vir die toekoms, want dit sal die wyn- of brandewynmaker meer beheer oor die uiteindelike produk gee en daardeur die wyn- of brandewynmaker help om 'n produk te vervaardig wat sy mark bevredig. 2012-08-27T11:35:13Z 2012-08-27T11:35:13Z 2002-12 Thesis http://hdl.handle.net/10019.1/52926 en_ZA Stellenbosch University 79 p. : ill. application/pdf Stellenbosch : Stellenbosch University
spellingShingle Brandy
Wine and wine making
Ethyl acetoacetate
Dissertations -- Wine biotechnology
Theses -- Wine biotechnology
Bayly, Jennifer Carr,1977-
Manipulating the levels of ethyl acetate and isoamyl acetate formation during the production of wine and brandy
title Manipulating the levels of ethyl acetate and isoamyl acetate formation during the production of wine and brandy
title_full Manipulating the levels of ethyl acetate and isoamyl acetate formation during the production of wine and brandy
title_fullStr Manipulating the levels of ethyl acetate and isoamyl acetate formation during the production of wine and brandy
title_full_unstemmed Manipulating the levels of ethyl acetate and isoamyl acetate formation during the production of wine and brandy
title_short Manipulating the levels of ethyl acetate and isoamyl acetate formation during the production of wine and brandy
title_sort manipulating the levels of ethyl acetate and isoamyl acetate formation during the production of wine and brandy
topic Brandy
Wine and wine making
Ethyl acetoacetate
Dissertations -- Wine biotechnology
Theses -- Wine biotechnology
url http://hdl.handle.net/10019.1/52926
work_keys_str_mv AT baylyjennifercarr1977 manipulatingthelevelsofethylacetateandisoamylacetateformationduringtheproductionofwineandbrandy