A Novel Method-a "Freeze-Blast" Method-to Disrupt Microbial Cells

概要

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A novel method to disintegrate cells by repidly blowing a frozen cell suspension at a high nitrogen gas flow against a target panel, that is, a freeze-blast method, was used on n-alkane-grown cells of Candida tropicalis pK 233 to extract several usuful substances. Electronmicroscopical observation that the yeast cells were broken into large fragments. Recovery of soluble enzymes(catalase and citrate synthase) with this method was comparable to that with the glass-beads and ultrasonication methods. A large part of membrane-associated enzymes (NADPH-cyrochrome c reductase, long-chain alcohol dehydrogenase, and ATPase) remained bound after disruption, and could be solubilized with 0.5%(w/v) Triton X-100. Under hypertonic condirions, mitochondria were found to be isolated directly without the pre-treatment of the yeast cells with a lytic enzyme. Agrose gel electrophoresis followed by Northern blot analysis showed that this method was also convenient for isolating RNAs form the cells. These results demonstrate that the freeze-blast method offers a novel technique to disrupt microbial cells, which might be applicable to other classes of cells.
社団法人日本生物工学会の論文
1989-01-25