Improving the Control of Wine Spoilage Micro-Organisms


Project Number
IWBT 05-04

Project title
Improving the control of wine spoilage micro-organisms

Project leader
Du Toit, M A

Institution
University of Stellenbosch. Department of Viticulture and Enology. Institute for Wine Biotechnology.

Team members
Van Rensburg, P A
Malherbe, D F
Morgan, J

Project description
The pediocin gene cassette was sub-cloned into an integration vector and the laboratory yeast strain is under evaluation at the moment. Lactic acid bacterial isolates from the South African winemaking environment were screened for anti-microbial peptide production; of the total isolates, 25 showed activity predominantly towards Lactobacillus and Pediococcus sensitive strains. Two isolates were selected for further characterization due to their stability in bacteriocin production.

The strains obtained using the Infruitec HEWL gene as template did not produce lysozyme when on an episomal plasmid, but produced zones of inhibition when integrated into the genome of the laboratory yeast strain. The strains obtained when using the HEWL gene obtained from Asselin et al showed activity when transformed with the episomal plasmid pJFH1(X), pJFH2(X) and pJFH4(X) as well as when these cassettes were integrated into the genome of lab yeast strains. The expression cassettes from pJFH2(X) and pJFH4(X) were sub- cloned into a vector suitable for integration into industrial strains and Vin13 has been successfully transformed with these plasmids. Studies are currently under way to determine whether expression of lysozyme takes place and also to create the final construct- pJFH3(X) so that a full set of transgenic yeast strains is available to proceed to the next objective. cc

The study using hydrogen sulphide as antimicrobial metabolite has resulted in progress towards the development of antimicrobial wine yeast starter culture strains for the production of wine with reduced levels of chemical preservatives and alcohol. Together with the other concurrent strategies of producing anti-microbial peptides and enzymes, the glucose oxidase-producing yeast might offer a viable way to meet consumer’s demands for decreased levels of sulphites.

Presentation(s)
Malherbe, D F, Van Rensburg, P, Du Toit, M A, Otero, C and Pretorius, I S. 2002. Expression of the Aspergillus niger glucose oxidase gene in Saccharomyces cerevisiae and its application in winemaking. Paper presented at the 22nd International Specialised Symposium on Yeasts. 25-28 March 2002. Pilanesberg, South Africa.

Malherbe, D F, Van Rensburg, P, Du Toit, M A, Otero, C and Pretorius, I S. 2002. Expression of the Aspergillus niger glucose oxidase gene in Saccharomyces cerevisiae and its application in winemaking. Presentation to the Cape Biotechnology Initiative.

Wassüng, R F, Du Toit, W J and Du Toit, M A. 2002. The influence of lysozyme on lactic acid bacteria, malolactic fermentation and the influence on the sensory quality of wine. Paper presented at the 26th National Congress of the South African Society for Enology and Viticulture. 13-15 November, Somerset West, South Africa.

Du Toit, M A. 2003. Bio-preservation and increasing the wholesomeness of wine. Presentation to the Lallemand Scientific Meeting: Beverages, fermentation, security and health. San Francisco, California, United States.

Du Toit, M A. 2003. Bio-preservation and lactic bacteria in winemaking. California Enological Research Association (CERA), Korbel Winery, Guernerville, California, United States.

Morgan, J, Vivier, M A, Pretorius, I S and Du Toit, M A. 2003. Screening, isolation and characterisation of antimicrobial and/or antifungal peptides produced by lactic acid bacteria isolated from wine. Paper presented at 1st FEMS Congress of European Microbiologists. 29 June – 3 July, Ljubljana, Slovenia.

Malherbe, D F, Du Toit, M A, Cordero Otero, R R, Pretorius, I S and Van Rensburg, P A. 2003. The development of a recombinant wine yeast strain capable of simultaneously reducing the alcohol content and bio-preserving wine. Paper presented at the 27th National Congress of the South African Society for Enology and Viticulture. 5-7 November, Somerset West, South Africa.

Morgan, J, Vivier, M A, Pretorius, I S and Du Toit, M A. 2004. Screening, isolation and characterisation of antimicrobial and / or antifungal peptides produced by lactic acid bacteria isolated from wine. Paper presented at the 12th Australian Wine Industry Technical Conference. Melbourne, Australia.

Malherbe, D F, Du Toit, M A, Cordero Otero, R R, Pretorius, I S and Van Rensburg, P A. 2004. Developing low-alcohol yeasts that are also able to bio-preserve wine. Paper presented at the 12th Australian Wine Industry Technical Conference. Melbourne, Australia.

Malherbe, D F, Du Toit, M A, Cordero Otero, R R, Pretorius, I S and Van Rensburg, P A. 2004. Low-alcohol yeasts that are able to bio-preserve wine. Paper presented at the 19th International Conference of the International Committee on Food Microbiology and Hygiene: Food Micro 2004. 12-16 September, Portoroza, Slovenia.

Malherbe, D F, Du Toit, M A, Cordero Otero, R R, Pretorius, I S and Van Rensburg, P A. 2004. The development of a recombinant wine yeast strain capable of simultaneously reducing the alcohol content and bio-preserving wine. Paper presented at the 13th Conference of the South African Society for Microbiology. 2-5 April, Stellenbosch, South Africa.

Du Toit, M A, Trollope, K M, Vivier, M A and Van Rensburg, P A. 2005. Enhancing the wholesomeness of wine. Paper presented at the 18th International Congress South African Association for Food Science and Technology(SAAFoST). 5-8 September, Stellenbosch, South Africa.

Malherbe, D F, Du Toit, M A, Cordero Otero, R R, Pretorius, I S and Van Rensburg, P A. 2004. The development of a recombinant wine yeast train capable of simultaneously reducing the alcohol content and bio-preserving wine. Paper presented at the 13th Conference of the South African Society for Microbiology. 2-5 April, Stellenbosch, South Africa.

Du Toit, M A, Trollope, K M, Vivier, M A and Van Rensburg, P A. 2005. Enhancing the wholesomeness of wine. Paper presented at the 18th Congress of the South African Association for Food Science and Technology. 5-8 September, Stellenbosch, South Africa.

Article
Pretorius, I, S, Du Toit, M A, Van Rensburg, P. 2003. Designer yeasts for the fermentation industry of the 21st century, Food Technology and Biotechnology, v. 41 (1) (p. 3-10)

Malherbe, D F. 2004. Let the yeast do the work for you: Less alcohol per bottle, WineLand, Mnth Jun (p. 105-106)

Du Toit, M, Vivier, M A, Van Rensburg, P. 2006. Enhancing the wholesomeness of wine, Food Review, v. 33 (2) (p. 25-26)

Schoeman, H. 2006. Is sulphur dioxide’s days numbered? Sulphur dioxide versus antimicrobial peptides in wine preservation, WineLand, v. 197 (p. 98-99)

Pamphlet
Du Toit, M, Vivier, M A, Van Rensburg, P. Biopreservation and increasing the wholesomeness of wine, IN: Beverages, fermentation and health. v. 11 (p. 27-32) Lallemand,

Paper
Morgan, J, Vivier, M A, Pretorius, I S, Du Toit, M. 2002. Screening, isolation and characterisation of antimicrobial, antifungal peptides produced by lactic acid bacteria isolated from wine, IN: Blair, R (ed). Proceedings of the 12th Australian Wine Industry Technical Conference. 24-29 July 2002, Melbourne, Australia. (p. 287) Australian Wine Research Institute, Adelaide, Australia.

Malherbe, D F, Du Toit, M, Cordero Otero, R R, Pretorius, I S, Van Rensburg, P. Blair, R (ed). 2004. Developing low-alcohol yeasts that are also able to biopreserve wine, IN: Blair, R (ed). Proceedings of the 12th Australian Wine Industry Technical Conference. 24-29 July 2004, Melbourne, Victoria. (p. 294-296) Australian Wine Research Institute, Adelaide, Australia.

FinalReport.pdf

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