The transformation of wine yeast with the Pediococcus acidilactici pediocin gene for control of spoilage bacteria


Project Number
IWBT 01-04

Project title
The transformation of wine yeast with the Pediococcus acidilactici pediocin gene for control of spoilage bacteria

Project leader
Du Toit, M A

Institution
University of Stellenbosch. Department of AgriSciences. Department of Viticulture and Oenology

Team members
Vivier, M A
D’Aguanno, V S
Schoeman, H
Krieling, S J
Morgan, J

Project description
The objectives were to express a functional bacteriocin peptide in Saccharomyces cerevisiae. This entailed the cloning of the pedA gene from a lactic acid bacterium Pediococcus acidilactici and the construction of yeast expression cassettes that will result in active bacteriocin peptides produced at high levels and secreted to the outside of the yeast cells. The expression levels, the activity and the nature of the recombinant bacteriocins were analysed to determine the usefulness of such an auto-sterilising yeast for the wine industry.

Bacteriocins of lactic acid bacteria have received considerable attention due to their potential application as natural preservatives. Bacteriocins may provide a valuable, additional and controllable tool for the inhibition of some deleterious wine-associated organisms. Although the use of sulphur dioxide cannot be eliminated, the levels of addition can be greatly reduced when used in conjunction with an antimicrobial peptide such as a bacteriocin produced from the fermenting organism itself.

Presentation(s)
Schoeman, H, Vivier, M A, Dicks, L M T and Pretorius, I S. 1996. Expression of the Pediococcus acidilactici PAC1.0 pediocin PA-1 gene in Saccharomyces cerevisiae. Paper presented at the 9th Conference of the South African Society for Microbiology. 8-10 July, Pretoria, South Africa.

Schoeman, H, Vivier, M A, Dicks, L M T and Pretorius, I S. 1997. Expression of the Pediococcus acidilactici PAC1.0 pediocin PA-1 gene in Saccharomyces cerevisiae. Paper presented at the 18th International Conference on Yeast Genetics and Molecular Biology. 31 March – 5 April, Stellenbosch, South Africa.

Schoeman, H, Du Toit, M A Vivier, M A, Dicks, L M T and Pretorius, I S. 1998. Development and potential application of wine yeast with antimicrobial activity. Paper presented at the 22nd National Congress of the South African Society for Enology and Viticulture. 12-13 November, Cape Town, South Africa.

Dicks, L M T, Schoeman, H, Vivier, M A and Pretorius, I S. 1998. Taxonomy of malolactic bacteria and biological control of spoilage micro-organisms in wine. Paper presented at the 32nd Vinitaly International Symposium. 16 April, Verona, Italy.

Pretorius, I S, Bauer, F F, Lambrechts, M G, Van Rensburg, P and Vivier, M A. 1998. Mögliche anwendugen von Genetisch verbesserten Weinhefen. Paper presented at the 7th International Symposium on Innovations in Enology and Intervitis Interfructa. 9-11 May 2004, Stuttgart, Germany.

Schoeman, H, Vivier, M A, Dicks, L M T and Pretorius, I S. 1998. Modification of wine yeasts to produce antimicrobial peptides (bacteriocins). Paper presented at Traditional Fermented Food Processing in Africa. Accra, Ghana.

Pretorius, I S, Schoeman, H, Du Toit, M A, Vivier, M A and Lambrechts, M G. 1999. Engineering wine yeast strains with antimicrobial activity. Paper presented at the 19th International Conference on Yeast Genetics and Molecular Biology. 25-30 May, Rimini, Italy.

Pretorius, I S, Schoeman, H, Du Toit, M A, Vivier, M A and Dicks, L M T. 1999. Engineering wine yeast strains with antimicrobial activity. Paper presented at 6th International Enology Symposium: OENO 99. 10-12 June 1999, Bordeaux, France.

Schoeman, H, Du Toit, M A, Vivier, M A, Dicks, L M T, Hastings, J W and Pretorius, I S. 1999. Development and potential applications of wine yeasts with antibacterial activity. Paper presented at the 24th World Congress of Vine and Wine and the 79th General Assembly of the OIV. 5-9 July 1999, Mainz, Germany.

Schoeman, H, Du Toit, M A, Vivier, M A, Dicks, L M T, Hastings, J W and Pretorius, I S. 1999. A yeast with antimicrobial activity as possible alternative to chemical preservatives. Paper presented at the 17th International Conference of the International Committee on Food Microbiology and Hygiene: FoodMicro 99. 13?17 September, Veldhoven, The Netherlands.

Schoeman, H, Vivier, M A, Du Toit, M A, Dicks, L M T and Pretorius, I S. 1999. Expression of pediocin PA-1, a bacteriocin produced by Pediococcus acidilactici PAC1.0 gene in Saccharomyces cerevisiae. Paper presented at 6th Symposium on Lactic Acid Bacteria: Genetics, Metabolism and Applications. 19-23 September, Veldhoven, The Netherlands.

Pretorius, I S, Bauer, F F, Du Toit, M A, Lambrechts, M G, Van Rensburg, P and Vivier, M A. 2000. Tailoring wine yeast for the third millennium: novel approach to the ancient art of winemaking. Paper presented at the 2nd International Congress of the South African Society for Enology and Viticulture. 8-10 November. Cape Town, South Africa.

Schoeman, H, Vivier, M A, Du Toit, M A, Dicks, L M T and Pretorius I S. 2000. Expression of pediocin PA-1, a bacteriocin produced by Pediococcus acidilactici PAC1.0 gene in Saccharomyces cerevisiae. Paper presented at the BIOY2K Combined Millennium Meeting. 23-28 January 2000, Grahamstown, South Africa.

D’Aguanno, V S, Schoeman, H, Du Toit, C, Du Toit, M A, Vivier, M A, Dicks, L M T, Hastings, J W and Pretorius, I S. 2000. Bio-preservation of wine using bactericidal wine yeast strains. Paper presented at the 2nd International Congress of the South African Society for Enology and Viticulture. 8-10 November, Cape Town, South Africa.

D’Aguanno, V S, Du Toit, C, Du Toit, M A and Pretorius, I S. 2001. Bio-preservation of wine with antimicrobial peptides. Paper presented at the 6th International Symposium on Innovations in Wine Technology: Intervitis Interfructa, Stuttgart, Germany.

D’Aguanno, V S, Du Toit, C, Du Toit, M A and Pretorius, I S. 2001. Development of bactericidal wine yeast starter cultures. Paper presented at the 20th International Conference on Yeast Genetics and Molecular Biology. 26-31 August, Prague, Czech Republic.

Du Toit, M A. 2001. Microbial spoilage and preservation of wine: Using weapons from nature’s own arsenal. Presentation to the Universidad de Castille – La Mancha, Ciudad Real, Spain.

Du Toit, M A, Du Toit, C, Krieling, S J and Pretorius, I S. 2001. Bio-preservation of wine with antimicrobial peptides and enzymes. Paper presented at the 26th World Congress of the Vine and Wine and the 81st General Assembly of the OIV. 11-17 October 2001, Adelaide, Australia.

Bauer, F F, Van Rensburg, P, Du Toit, M A and Pretorius, I S. 2001. The yeast biotechnology programme at the Institute for Wine Biotechnology. Paper presented at the Cape Biotechnology Forum 2001. Cape Town, South Africa.

Du Toit, M A. 2001. Lactic acid bacteria and spoilage. Presentation at the Seminar on Wine bacteria: Winemakers’ friend or enemy’. Lecture. University of Stellenbosch, Department of Viticulture and Enology. Stellenbosch, South Africa.

Du Toit, M A, Du Toit, C, Krieling, S J and Pretorius, I S. 2001. Bio-preservation of wine with antimicrobial peptides. Paper presented at the 25th National Congress of the South African Society for Enology and Viticulture. 15-16 November, Cape Town, South Africa.

Du Toit, M A, Du Toit, C, Van Rensburg, P and Pretorius, I S. 2002. The application of nisin, pediocin and leucocin as bio-preservatives in wine. Paper presented at the 12th Conference of the South African Society for Microbiology and the 3rd International Virology and Microbiology Conference. 2-5 April, University of the Free State, Bloemfontein, South Africa.

Pretorius, I S, Bauer, F F, Du Toit, M A, Van Rensburg, P and Vivier, M A. 2002. The genetic improvement of wine yeast starter cultures. Paper presented at the 13th International Symposium of Enology. Montpellier, France.

Morgan, J, Du Toit, M, Vivier, M A and Pretorius, I S. 2002. Screening, isolation and characterisation of antimicrobial peptides produced by lactic acid bacteria isolated from wine. Paper presented at the 12th Conference of the South African Society for Microbiology. 2-5 April, Bloemfontein, South Africa.

Article
Schoeman, H, Vivier, M A, Du Toit, M A, Dicks, L M T, Pretorius, I S. 1999. The development of bactericidal yeast strains by expressing the Pediococcus acidilactici pediocin gene (pedA) in Saccharomyces cerevisiae, Yeast, v. 15 (8) (p. 647-656)

Du Toit, M A, Pretorius, I S. 2000 Special issue. Microbial spoilage and preservation of wine: using weapons from nature’s own arsenal – a review, South African Journal of Enology and Viticulture, v. 21 (p. 74-96)

Du Toit, M A, Du Toit, C, Krieling, S J, Pretorius, I S. 2002. Biopreservation of wine with antimicrobial peptides, Bulletin d’OIV, Mnth May v. 75 (855-856) (p. 284-302)

FinalReport.pdf

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