Optimisation of Saccharomyces cerevisiae BRYC 501 ascospore formation and recovery for heat inactivation experiments

Abstract

Why was the work done: Ascospores from Saccharomyces cerevisiae BRYC 501 are useful biological indicators for validating the pasteurisation of beer. Ascospores are formed by yeast in response to low nutrient conditions and are characterised by greater resistance to stressors including desiccation, freezing and, during pasteurisation, high temperatures. Accordingly, the sporulation temperature of yeast ascospores, their shelf life, heat resistance and recovery need to be investigated. How was the work done: The role of temperature in sporulation was investigated at 25, 27 and 30°C by measurement of the rate of sporulation over ten days. Shelf life of ascospores was established in deionised water at 1-5°C over 120 days by the measurement of heat resistance in McIlvaine's buffer. The recovery conditions of ascospores were determined using six agars after heat inactivation experiments in McIlvaine's buffer, alcohol free and lager beer. What are the main findings: The sporulation rate of S. cerevisiae ascospores was highest at 27°C, and lowest at 30°C. Heat resistance of ascospores formed at 30°C was low with a D60 value of <3 minutes, whereas at 25 and 27°C the D60 was 8.8 and 9.2 minutes. The shelf life of ascospores was 60 days after an initial 14 days of maturation at both 25 and 27°C. The variation of heat resistance for ascospores grown at 25°C was shown to be lower than those grown at 27°C. Use of yeast extract-peptone-glucose agar to recover heat injured cells was > 200% higher than with the control agar. Why is the work important: Sporulation should be performed at 25°C to produce the maximum number of spores for heat inactivation experiments. Spores can be maintained for 60 days, enabling several experiments to be performed. After heat inactivation experiments, ascospores should be recovered on YEPG agar for 10 days at 27°C