Study of physical and biological factors involved in the disruption of E. coli by hydrodynamic cavitation.
Hydrodynamic cavitation results in flow restriction in a flow system causing rapid pressure fluctuations and significant fluid forces. These can be harnessed to mediate microbial cell damage. Hydrodynamic cavitation was studied for the partial disruption of E. coli and selective release of specific proteins relative to the total soluble protein. The effects of the cavitation number, the number of passes, and the specific growth rate of E. coli on the release of periplasmic and cytoplasmic proteins were studied. At the optimum cavitation number of 0.17 for this experimental configuration, 48% of the total soluble protein, 88% of acid phosphatase, and 67% of beta-galactosidase were released by hydrodynamic cavitation in comparison with the maximum release attained using multiple passes through the French Press. The higher release of the acid phosphatase over the total soluble protein suggested preferred release of periplasmic compounds. This was supported by SDS-PAGE analysis. The absence of micronization of cell material resulting in the potential for ease of solid-liquid separation downstream of the cell disruption operation was confirmed by TEM microscopy. E. coli cells cultivated at a higher specific growth rate (0.36 h(-1)) were more easily disrupted than slower grown cells (0.11 h(-1)). The specific activity of the enzyme of interest released by hydrodynamic cavitation, defined as the units of enzyme in solution per milligram of total soluble protein, was greater than that obtained on release by the French Press, high-pressure homogenization, osmotic shock, and EDTA treatment. The selectivity offered indicates the potential of enzyme release by hydrodynamic cavitation to ease the purification in the subsequent downstream processing.
Balasundaram
B
B
Department of Chemical Engineering, University of Cape Town, Rondebosch 7701, South Africa.
Harrison
S T L
ST
D000135:Acid Phosphatase / Q000737:chemistry / Q000378:metabolism*; D001433:Bacteriolysis / Q000502:physiology*; D002478:Cells, Cultured; D003593:Cytoplasm / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry* / Q000254:growth & development / Q000378:metabolism*; D007700:Kinetics; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D001616:beta-Galactosidase / Q000737:chemistry / Q000378:metabolism*
D011506:Proteins; D000135:Acid Phosphatase; D001616:beta-Galactosidase