Chitin synthesis in response to environmental stress

Pauw, Marina (2014-04)

Thesis (MSc)--Stellenbosch University, 2014.

Thesis

ENGLISH ABSTRACT: Previous studies have indicated that fermentation with yeast strains whose cell walls contain higher chitin levels may lead to reduced wine haze formation. In order to adjust cell wall chitin levels, more information on the regulation of chitin synthesis in wine-relevant yeast is required. Yeast cells are known to increase chitin levels when subjected to certain environmental changes such as an increase in temperature. The main aim of this project was to investigate chitin accumulation and synthesis in wine yeast strains when exposed to environmental change. This was achieved by subjecting the strains to various environmental conditions and comparing chitin levels. The information gained may aid future selection and/or manipulation of yeast strains for the production of higher chitin levels. Three Saccharomyces cerevisiae strains and two Saccharomyces paradoxus strains were subjected to conditions that had been linked to a change in chitin synthesis in past studies in laboratory yeast strains. Of the conditions used in this study, the addition of calcium to a rich media led to the highest cell wall chitin levels. The data also show that chitin synthesis is largely strain dependant. Two conditions which resulted in increased chitin deposition were chosen for gene expression analyses, using strains with strongly diverging average chitin levels. Results showed that an increase in chitin levels correlates with an increase in expression of GFA1, the gene encoding for the first enzyme of the chitin synthesis pathway. Overall, this study provides novel insights into chitin synthesis in Saccharomyces cerevisiae wine yeast strains as well as Saccharomyces paradoxus strains, with possible future implications on haze prevention studies.

AFRIKAANSE OPSOMMING: Vorige studies het aangetoon dat fermentasie met gisrasse waarvan die selwande hoë chitienvlakke bevat, kan lei tot verminderde wynwaasvorming. Om selwandchitienvlakke aan te pas, word daar meer inligting rakende die regulering van chitienvlakke in wyn gisrasse verlang. Dit is bekend dat gisselle chitienvlakke verhoog wanneer die selle onderwerp word aan sekere veranderinge in die omgewing soos ’n verhoging in temperatuur. Die hoofdoel van hierdie projek was om die chitienopbou en -sintese in wyngisrasse te ondersoek waar gis blootgestel word aan omgewingsveranderinge. Dit is bereik deur die selle aan verskeie omgewingstoestande bloot te stel en chitienvlakke met mekaar te vergelyk. Die inligting hieruit verkry kan toekomstige gisraskeuses asook die manipulering van gisrasse met die oog op hoër vlakke van chitienproduksie vergemaklik. Drie Saccharomyces cerevisiae rasse en twee Saccharomyces paradoxus rasse is onderwerp aan toestande wat in vorige studies gekoppel is aan ’n verandering in chitienvorming in laboratorium-gisrasse. Van die toestande toegepas in hierdie studie, het die toevoeging van kalsium tot ’n nutrientryke medium gelei tot die hoogste chitienvlakke in selwande. Die data toon ook aan dat chitiensintese hoofsaaklik rasverwant is. Twee toestande wat gelei het tot verhoogde chitienafsetting is gekies vir geen-uitdrukkingsanalise, terwyl rasse gebruik is met gemiddelde chitienvlakke wat wyd uiteenlopend is. Die resultate het getoon dat ’n verhoging in chitienvlakke ooreenstem met ’n verhoging in die uitdrukkingsvlakke van GFA1, die geen wat kodeer vir die eerste ensiem in die chitiensintesebaan. Oor die algemeen verskaf hierdie studie nuwe insigte oor chitiensintese in Saccharomyces cerevisiae wyngisrasse en Saccharomyces paradoxus rasse en verskaf dit belangrike inligting vir moontlike toekomstige studies oor waasvoorkoming.

Please refer to this item in SUNScholar by using the following persistent URL: http://hdl.handle.net/10019.1/86435
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