Disruption of Brewers' yeast by hydrodynamic cavitation: Process variables and their influence on selective release.
Intracellular products, not secreted from the microbial cell, are released by breaking the cell envelope consisting of cytoplasmic membrane and an outer cell wall. Hydrodynamic cavitation has been reported to cause microbial cell disruption. By manipulating the operating variables involved, a wide range of intensity of cavitation can be achieved resulting in a varying extent of disruption. The effect of the process variables including cavitation number, initial cell concentration of the suspension and the number of passes across the cavitation zone on the release of enzymes from various locations of the Brewers' yeast was studied. The release profile of the enzymes studied include alpha-glucosidase (periplasmic), invertase (cell wall bound), alcohol dehydrogenase (ADH; cytoplasmic) and glucose-6-phosphate dehydrogenase (G6PDH; cytoplasmic). An optimum cavitation number Cv of 0.13 for maximum disruption was observed across the range Cv 0.09-0.99. The optimum cell concentration was found to be 0.5% (w/v, wet wt) when varying over the range 0.1%-5%. The sustained effect of cavitation on the yeast cell wall when re-circulating the suspension across the cavitation zone was found to release the cell wall bound enzyme invertase (86%) to a greater extent than the enzymes from other locations of the cell (e.g. periplasmic alpha-glucosidase at 17%). Localised damage to the cell wall could be observed using transmission electron microscopy (TEM) of cells subjected to less intense cavitation conditions. Absence of the release of cytoplasmic enzymes to a significant extent, absence of micronisation as observed by TEM and presence of a lower number of proteins bands in the culture supernatant on SDS-PAGE analysis following hydrodynamic cavitation compared to disruption by high-pressure homogenisation confirmed the selective release offered by hydrodynamic cavitation.
Department of Chemical Engineering, University of Cape Town, Rondebosch 7701, South Africa.
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B
Balasundaram
S T L
ST
Harrison
D000426:Alcohol Dehydrogenase / Q000378:metabolism*; D002458:Cell Fractionation / Q000379:methods*; D002473:Cell Wall / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D006026:Glycoside Hydrolases / Q000378:metabolism*; D019897:Periplasm / Q000201:enzymology; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000648:ultrastructure
D000426:Alcohol Dehydrogenase; D006026:Glycoside Hydrolases