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Separation of the Prodigiosin‐Localizing Crude Vesicles Which Retain the Activity of Protease and Nuclease in Serratia marcescens
Author(s) -
Kobayashi Nobuharu,
Ichikawa Yoichi
Publication year - 1991
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1991.tb01592.x
Subject(s) - lysis , prodigiosin , biology , vesicle , serratia marcescens , protease , biochemistry , differential centrifugation , chromatography , centrifugation , nuclease , distilled water , microbiology and biotechnology , enzyme , chemistry , escherichia coli , membrane , gene
Crude vesicles in which prodigiosin is localized were separated from pigmented Serratia marcescens. The bacteria were grown on peptonegiycerol agar plate, suspended in saline, and fractionated into cells, vesicles, and supernatant by differential centrifugation. Electron microscopic observations showed that the fractionation was conducted properly and the separated vesicles were lysed in distilled water. The vesicles suspended in saline retained 100 kilodalton protein of which amount is correlated with prodigiosin level, but the 100 kDa protein was found in the supernatant when the vesicles were lysed in distilled water. The vesicle fraction retained few colony‐forming units and little detectable activity of NADH oxidase, but showed much higher activities of protease and nuclease than the cell fraction. The profiles of the activities of the protease and the nuclease in the fractions were different from each other, that is, the protease activity in the vesicle fraction was lower than that in the supernatant fraction, whereas the nuclease activity in the vesicle fraction was higher than that in the supernatant fraction, suggesting that the two extracellular enzymes were released from the pigmented bacteria by different mechanisms.