An extracellular proteolytic enzyme from Scopulariopsis brevicaulis. 1. Purification and properties

Abstract

A proteolytic enzyme present in culture filtrates of Scopulariopsis brevicaulis was purified about 200-fold by (NH4)2SO4 and ethanol fractionations followed by chromatography on DEAE-cellulose, DEAE-Sephadex, and hydroxylapatite. Ultracentrifugation of the purified enzymes showed only one sedimenting component and its molecular weight was estimated to be about 24 000. The protease hydrolyzed casein, urea-denatured hemoglobin, gelatin, fibrinogen, fibrin, insulin chains A and B, but not human serum albumin or ovalbumin. It also coagulated milk. The enzyme had no action on the various peptides tested and showed low esterase activity. Optimum pH for casein hydrolysis was 10.5 to 11; for hemoglobin hydrolysis 7.0–9.5, and for gelatin hydrolysis, 6.0–8.0. The enzyme activity was unaffected by most metal ions, SH-reagents, and some natural trypsin inhibitors. The protease was strongly inhibited by diisopropylfluorophosphate and phenylmethanesulfonyl fluoride. Although similar in some respects to CA-7, the enzyme isolated from Aspergillus oryzae, and other alkaline proteases, the S. brevicaulis protease does not appear to be identical with any one of them.

Production and properties of fibrinolytic enzyme in solid state cultures of Fusarium pallidoroseum

“…Singh and Vezina (1971) observed the presence of some caseinase activity associated with fibrinolytic enzyme of Sropulariopsis It was also noticed that the enzyme was relatively sensitive to heat treatments in absence of its s&&rate, since it lost about 70% of its activity when exposed to 55°C for 15 min. Maximum enzyme activity was recorded atpH 7 and the enzyme was fairly stable in a-pH range of 6.5 to 9.0 retaining 80% of its activity.…”

Purification and some properties of pure Cochliobolus lunatus fibrinolytic Enzyme

“…The fibrinolytic enzyme present in culture filtrate of Scopulariopsis brevicaulis was purified by chromatography on DEAE-Sephadex and hydroxylapatite. 3 Tomoda et a14 purified Fusarium alkaline protease from the culture broth through a sequence of chromatography on Asmit resin, DEAE-cellulose, CM-Sephadex, and gel filtration on Sephadex G-75.…”