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Subcellular fractionation of the Gram negative rumen bacterium, Butyrivibrio S 2 , by protoplast formation, and localisation of lipolytic enzymes in the plasma membrane
Author(s) -
Hazlewood G.P.,
Cho K.Y.,
Dawson R.M.C.,
Munn E.A.
Publication year - 1983
Publication title -
journal of applied bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 0021-8847
DOI - 10.1111/j.1365-2672.1983.tb01330.x
Subject(s) - protoplast , lysis , lysozyme , differential centrifugation , membrane , centrifugation , biochemistry , chromatography , cell wall , cell fractionation , biology , chemistry
Treatment of the anaerobic, Gram negative general fatty acid auxotroph Butyrivibrio S2 with lysozyme in low molarity buffers resulted in the formation of protoplasts, some of which retained the original rod‐shaped morphology of the organism. The protoplasts were stabilised by the presence of Mg 2+ ions. Most of the phospholipase A and C and galactolipase activity of the cells was retained by the protoplasts. Electron microscopy and chemical markers were used to monitor the separation of plasma membrane and cell wall fragments by density gradient centrifugation after osmotic lysis of protoplasts. Phospholipase and galactolipase activities were demonstrated in a subcellular fraction which contained only fragments of plasma membrane.