Abstract Extra-cellular proteinase of Pseudomonas aeruginosa IFO 3080 ★ has been obtained as needle-like crystals by the usual purification procedures, and some of its general properties have been examined. 1. 1. The proteinase after recrystallization three times, appears to be homogeneous on electrophoresis and ultracentrifugation; a small amount of impurity has been detected by column chromatography on DEAE-cellulose. 2. 2. The iso-electric point has been found to be at a pH slightly below 4.08. Maximum absorption of the ultraviolet spectrum is at 280 mμ and the minimum at 250 mμ. 3. 3. The proteinase is fairly stable over the pH range of 5 to 9 at 30°, and, in neutral solution, below 50°. 4. 4. The optimum pH for proteolytic activity is pH 7–9, and the optimum temperature is 60°. 5. 5. The proteinase acts on various protein substrates such as casein, gelatin, egg albumin and haemoglobin, but not on native collagen. Various synthetic substrates such as benzoyl- l -arginine amide, acetyl- l -tyrosine ethyl ester, carbobenzoxy- l -glutamyl- l -tyrosine, carbobenzoxyglycyl- l -phenylalanine and l -leucinamide are not hydrolysed. 6. 6. Reducing agents, -SH reagents, DFP and natural inhibitors from soy bean and potato do not affect the enzymic activity of the proteinase. Oxidizing agents inactivate the enzymic activity. Further, metal-complexing agents such as EDTA and o -phenanthroline inhibit the enzymic activity, although α , α ′-dipyridyl, NaF, citrate and oxalate do not affect it. Heavy-metal ions such as Ag + , Cu 2+ , Hg 2+ , Pb 2+ , Fe 2+ , etc., inactivate the enzymic activity. Inactivation is also seen with high concentrations of urea.
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