Effect of moisture content on the invertase activity of freeze-dried S. cerevisiae.
The invertase activity of intact Saccharomyces cerevisiae submitted to freezing-thawing was affected by pH, the chemical nature of the buffer, and the freezing cooling rate (CR), leading in some cases to a complete invertase inactivation (acetate buffer, pH 4.0, CR = 0.5 degree C/min). Once established under adequate freezing conditions the invertase activity remained unchanged after freeze-drying. Nevertheless, in some cases the cell-growing capability after freeze-drying diminished around 70%, mainly if the frozen cell suspension was attained through freezing carried out at CR = 0.5 degree C/min. Water sorption isotherms of freeze-dried samples (freeze-dryer Edwards L-4KR; 30 degrees C and 0.1 mB) were determined at 10 and 25 degrees C. The monolayer moisture content (MMC) at each temperature (12.7 and 3.71 for 10 and 25 degrees C, respectively) was calculated from isotherms by applying BET and GAB models. Freeze-dried yeast with water activity (Aw) between 0 and 0.33 (about the MMC value) maintained at 25 degrees C for 235 days and at 89 degrees C for 15 min retained at least 85% of its original invertase activity (IA), whereas samples with Aw > MMC lost at least 60% of its IA. X ray diffraction showed that the freeze-dried cake before and after storage presented an amorphous structure.
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Faculdade de Ciências Farmacêuticas, University of São Paulo, Brasil.
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D002021:Buffers; D004795:Enzyme Stability; D005069:Evaluation Studies as Topic; D005612:Freeze Drying / Q000379:methods*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000201:enzymology / Q000254:growth & development; D014867:Water / Q000032:analysis; D014961:X-Ray Diffraction; D043324:beta-Fructofuranosidase
D002021:Buffers; D014867:Water; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase