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Investigating the role of Brettanomyces and Dekkera during winemaking

Thesis (PhD (Genetics. Plant Biotechnology))--Stellenbosch University, 2008.

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Main Author: Oelofse, Adriaan
Other Authors: Du Toit, M.
Format: Thesis
Language:English
Published: Stellenbosch : Stellenbosch University 2008
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access_status_str Open Access
author Oelofse, Adriaan
author2 Du Toit, M.
author_browse Du Toit, M.
Oelofse, Adriaan
author_facet Du Toit, M.
Oelofse, Adriaan
author_sort Oelofse, Adriaan
collection Thesis
dc_rights_str_mv Stellenbosch University
description Thesis (PhD (Genetics. Plant Biotechnology))--Stellenbosch University, 2008.
format Thesis
id oai:scholar.sun.ac.za:10019.1/1239
institution Stellenbosch University (South Africa)
language English
last_indexed 2026-06-10T12:43:27.297Z
license_str Other — see source repository
provenance_str_mv Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository
publishDate 2008
publishDateRange 2008
publishDateSort 2008
publisher Stellenbosch : Stellenbosch University
publisherStr Stellenbosch : Stellenbosch University
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source_str SUNScholar — Stellenbosch University Repository
spelling oai:scholar.sun.ac.za:10019.1/1239 Investigating the role of Brettanomyces and Dekkera during winemaking Oelofse, Adriaan Du Toit, M. Pretorius, I. S. Lonvaud-Funel, A. Stellenbosch University. Faculty of AgriSciences. Dept. of Genetics. Institute for Plant Biotechnology . Brettanomyces Wine spoilage Volatile phenols Wine and wine making Wine microbiology Dekkera Yeast Dissertations -- Plant biotechnology Theses -- Plant biotechnology Thesis (PhD (Genetics. Plant Biotechnology))--Stellenbosch University, 2008. Wine quality is greatly influenced by the number of microorganisms, which occur throughout the winemaking process. These microorganisms are naturally present on the grapes and in the cellar from where they can be introduced to the winemaking process at any given time and consequently impart specific contributions to the wine quality. However, these microorganisms can be seen either as beneficial or as wine spoilage microorganisms, depending on the conditions under which they can proliferate during the winemaking process. Wine yeasts (Saccharomyces spp.) are typically responsible for the alcoholic fermentation; lactic acid bacteria (LAB) are responsible for malolactic fermentation (MLF), while acetic acid bacteria (AAB) and other wild yeasts (non-Saccharomyces spp.) are typically associated with the formation of off-flavours under poorly controlled winemaking conditions. In recent years, evidence from the wine industry has highlighted a specific group of non-Saccharomyces yeast species as a serious cause for wine spoilage that required more research investigations. Yeast of the genus Brettanomyces or its teleomorph Dekkera has been identified as one of the most controversial spoilage microorganisms during winemaking as they can produce several compounds that are detrimental to the organoleptic quality of wine. This has triggered the research initiative behind this doctoral study on the significance of Brettanomyces and Dekkera yeasts during winemaking. In this dissertation, various aspects of the detection, isolation and identification methods of Brettanomyces yeast from the winemaking environment were investigated. As a first objective, a culture collection of Brettanomyces bruxellensis wine isolates had to be established. This followed after the isolation of Brettanomyces yeasts from various red wine cultivars from South African wineries from different stages of the winemaking process. Different conventional microbiological methods such as plating on selective agar media and microscopy were investigated along with molecular identification techniques such as the polymerase chain reaction (PCR) in this regard. Other focus areas of this study aimed at performing genetic characterisation and differentiation studies of B. bruxellensis wine isolates. For this purpose, different intraspecific identification methods were investigated on several strains, including strains of European origin. The application of molecular techniques allowing strain identification aided in the selection of specific strains that were evaluated for volatile phenol production in synthetic media and wine. The results obtained from this work indicated that a large degree of genetic diversity exists among B. bruxellensis strains and that the volatile phenol production differed between the strains after evaluation in synthetic media and wine. In addition to the molecular intraspecific strain identification techniques that were investigated, a feasibility study was also performed that focused on evaluating Fourier transform infrared (FTIR) spectroscopy combined with chemometrics as an alternative approach for differentiating between B. bruxellensis strains. The two approaches of FTIR spectroscopy that were investigated involved the use of firstly, Fourier transform mid-infrared (FTMIR) spectroscopy to obtain spectral fingerprints of spoiled wines by different B. bruxellensis strains; and secondly, Attenuated total reflectance (FTIR-ATR) to obtain spectral fingerprints from whole cells of B. bruxellensis on microbiological agar media. The results of this study illustrated the potential of FTIR spectroscopy to become a reliable alternative to molecular based methods for differentiating between B. bruxellensis strains and for characterisation studies. The formation of volatile phenols in wine by species of the genera Brettanomyces and Dekkera is one of the primary reasons for their classification as wine spoilage yeasts. The enzymatic activities of this reaction have been identified and involve a phenyl acrylic (phenolic) acid decarboxylase (PAD) and a vinyl phenol reductase (VPR). However, only a limited amount of information is available about these enzymes from Brettanomyces/Dekkera yeasts and no genetic data have been described. It was therefore imperative that this dissertation should include a genetic investigation into the phenylacrylic (hydroxycinnamic) acid decarboxylase from the species B. bruxellensis involved in the formation of volatile phenols. Strategies that were investigated included various molecular DNA techniques and protein purification procedures to obtain either genetic or protein sequence data. The decarboxylase activity of this yeast species towards p-coumaric acid was demonstrated and substantial genetic sequence data was obtained. The results from this dissertation made a substantial contribution to the current available knowledge about Brettanomyces/Dekkera spp. and led to a better understanding of this wine spoilage yeast. This research developed a platform from which further investigations could follow and the knowledge gained will be invaluable for future Brettanomyces research projects at the Institute for Wine Biotechnology at Stellenbosch University. Doctoral 2008-11-25T13:12:29Z 2010-06-01T08:16:13Z 2008-11-25T13:12:29Z 2010-06-01T08:16:13Z 2008-12 Thesis http://hdl.handle.net/10019.1/1239 en Stellenbosch University application/pdf Stellenbosch : Stellenbosch University
spellingShingle Brettanomyces
Wine spoilage
Volatile phenols
Wine and wine making
Wine microbiology
Dekkera
Yeast
Dissertations -- Plant biotechnology
Theses -- Plant biotechnology
Oelofse, Adriaan
Investigating the role of Brettanomyces and Dekkera during winemaking
title Investigating the role of Brettanomyces and Dekkera during winemaking
title_full Investigating the role of Brettanomyces and Dekkera during winemaking
title_fullStr Investigating the role of Brettanomyces and Dekkera during winemaking
title_full_unstemmed Investigating the role of Brettanomyces and Dekkera during winemaking
title_short Investigating the role of Brettanomyces and Dekkera during winemaking
title_sort investigating the role of brettanomyces and dekkera during winemaking
topic Brettanomyces
Wine spoilage
Volatile phenols
Wine and wine making
Wine microbiology
Dekkera
Yeast
Dissertations -- Plant biotechnology
Theses -- Plant biotechnology
url http://hdl.handle.net/10019.1/1239
work_keys_str_mv AT oelofseadriaan investigatingtheroleofbrettanomycesanddekkeraduringwinemaking