Masters Degrees (Viticulture and Oenology)
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Browsing Masters Degrees (Viticulture and Oenology) by Subject "Acetobacter"
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- ItemIsolation and characterisation of the antimicrobial peptides produced by acetic acid bacteria(Stellenbosch : Stellenbosch University, 2003-03) Oelofse, Adriaan; Du Toit, M.; Pretorius, I. S.; Lambrechts, M. G.; Stellenbosch University. Faculty of AgriSciences. Dept. of Viticulture and Oenology. Institute for Wine Biotechnology.ENGLISH ABSTRACT: Wine quality is greatly influenced by the number of microorganisms, which occur throughout the winemaking process. Yeasts are responsible for the alcoholic fermentation, the lactic acid bacteria (LAB) are responsible for malolactic fermentation (MLF), while acetic acid bacteria (AAB) are responsible for converting ethanol to acetic acid. These microorganisms are present on the grapes and in the cellar and these consequently serve as gateways to the fermentation tanks where they will affect the wine quality. However, these microorganisms can be seen either as beneficial or as wine spoilage microorganisms, depending on the conditions that prevail throughout the winemaking process. It is thus very important to prevent any process that could lead to the lowering of the wine quality. In this regard, some of the factors that should always be evaluated include the quality of the grapes, winemaking techniques and quality control. One of the measures that have been implemented during winemaking to ensure the microbial stability is the use of chemical preservatives. Sulphur dioxide (502) has been, and is, used widely as primary preservative in winemaking. However, an ever-increasing consumer resistance against the use of chemical preservatives has developed as it poses possible health risks and decreases the sensorial quality of wine. An alternative approach to chemical preservation that has triggered numerous new investigations, is biological preservation or biopreservation. This is the use of the natural microbial flora and/or their antimicrobial products, such as bacteriocins, to inhibit or destroy the other sensitive microorganisms that are unwanted in the same environment. Evidence in the wine industry has shown that bacterial spoilage still is a very common problem in many wineries. This bacterial spoilage can lead to, amongst other, two main problems, which are of great concern to winemakers. This include high levels of volatile acidity, resulting in the wine having a vinegary off-flavour, and sluggish/stuck fermentation, which is the result of compounds such as acetic- and other fatty acids that causes inhibition of the yeast's growth. With acetic acid being the common link in both cases, it became evident that investigations should be performed on the main producer of acetic acid, namely AAB. As a result, AAB turned out to be one of the main spoilage microorganisms associated with winemaking. Most of the research on biopreservation in the food and beverage industry has been performed on the Gram-positive LAB. The fact that their spectrum of inhibition currently excludes most Gram-negative bacteria, specifically AAB, indicated that AAB should be screened in search of possible antimicrobial compounds that could be applied to control their cell numbers during winemaking. No evidence of antimicrobial action amongst AAB could be found in literature, therefore this work was considered novel. The main objectives of this study were to screen wine isolates of AAB for the production of antimicrobial compounds. This was followed by the isolation and preliminary characterisation of the antimicrobial substances produced. Various attempts to optimise the production of the antimicrobial compounds and isolation procedures, were also included. This study forms part of a larger research programme that has been initiated at the Institute for Wine Biotechnology at Stellenbosch University on the biopreservation in wine. Our results indicated that possible antimicrobial compounds of proteinaceous nature, produced by AAB isolated from wine, do exist. It was found that two different species of AAB, namely Acetobacter aeeti and Gluconobacter frateurii, produced antimicrobial compounds that inhibited other species of AAB. Preliminary results indicated that these compounds are heat sensitive and stable in a wide pH range. It was also shown that after the action of proteolytic enzymes, such as proteinase K and a-chemotrypsin, all inhibitory activity was lost. This study also revealed the existence of the species Gluconobacter frateurii, which have not yet been associated with the winemaking environment. This study made a valuable contribution to the limited amount of information and understanding of AAB, not only in the wine environment, but also elsewhere. The results and findings of this research would serve as platform for further projects. This might soon lead to the development of antimicrobial substances or tailored wine-yeasts with antimicrobial abilities, which can be applied during winemaking to assist the winemaker in combatting high cell numbers and subsequent spoilage by AAB.