Relationship between intracellular protein denaturation and irreversible inactivation of Saccharomyces pastorianus by...

To clarify the relationship between irreversible inactivation and intracellular protein denaturation of Saccharomyces pastorianus by low-pressure carbon dioxide microbubbles (CO₂ MB) treatment, a storage test of S. pastorianus cells treated with CO₂ MB was performed, and the effect on the intracellular protein was investigated. In the storage test, the S. pastorianus population, which decreased below the detection limit by CO₂ MB treatment at a temperature of 45 and 50°C (MB45 and MB50), and thermal treatment at a temperature of 80°C (T80), remained undetectable during storage for 3 weeks at 25°C. However, 4.1 and 1.3-logs of the S. pastorianus populations, which survived after CO₂ MB treatment at temperatures of 35 and 40°C (MB35 and MB40), increased gradually during storage for 3 weeks at 25°C. Insolubilization of intracellular proteins in S. pastorianus increased with increasing the temperature of CO₂ MB treatment. Activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) identified as one of the insolubilized proteins increased at MB35 and MB40 than non-treatment but disappeared at MB45 and MB50, and T80. Therefore, it was revealed that S. pastorianus cells inactivated below the detection level by CO₂ MB treatment did not regrow and that the denaturation of intracellular proteins of S. pastorianus was caused by CO₂ MB and thermal treatments. Furthermore, it was suggested that denaturation of intracellular vital enzymes was an important factor for achieving irreversible inactivation of S. pastorianus by CO₂ MB and thermal treatments.