Proteinase enzyme system of lactic streptococci. II. Role of membrane proteinase in cellular function.
The effect of several environmental conditions on the structure and activity of a membrane-associated proteinase from Streptococcus lactis was investigated. The activity of the enzyme varied with pH. Before storage at 3 C, maximal activity occurred at pH 6.0, but was minimal at this pH after storage. At all pH values tested, the enzyme was inactivated after storage. After storage at 3 C, the enzyme showed gross structural alterations with a concomitant loss of activity. Gel filtration and sedimentation velocity data indicated that inactivation of the enzyme was the result of aggregation to higher molecular weight forms. p-Hydroxymercuribenzoate prevented inactivation of the enzyme during storage by preventing aggregation. Activity was correlated with disaggregation of polymer forms of the enzyme to an active monomer. The storage-inactivated enzyme could be reactivated by treatment of the enzyme with cysteine, glutathione, or ferrous ion. Glutathione enabled stored cells to produce acid at their original rate when subcultured in milk. This was attributed to the effect of glutathione on the membrane proteinase. The data suggested that the biological activity of stored cells may be dependent upon the activity of the membrane proteinase.
D000818:Animals; D001565:Benzoates / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects* / Q000201:enzymology*; D002850:Chromatography, Gel; D003470:Culture Media; D003545:Cysteine / Q000494:pharmacology; D005978:Glutathione / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000494:pharmacology; D007773:Lactates / Q000096:biosynthesis; D008628:Mercury; D008892:Milk; D010447:Peptide Hydrolases / Q000378:metabolism*; D013291:Streptococcus / Q000201:enzymology*; D013696:Temperature; D014461:Ultracentrifugation
D001565:Benzoates; D003470:Culture Media; D007773:Lactates; D007501:Iron; D010447:Peptide Hydrolases; D008628:Mercury; D005978:Glutathione; D003545:Cysteine