[Partial purification and properties of protease from Torula thermophila].
11-Fold purified protease preparation is isolated from cultural medium of Torula thermophila UzPT-1 by means of ammonium sulphate precipitation and gel chromatography through Sephadex G-100. Disc polyacrylamide gel electrophoresis revealed two portease components, one of them possessing proteolytic activity. pH interval for protease activity was found to be 3.5-12, the maximal activity was observed at pH 8.5-11, the highest enzyme resistance--at pH 6-8. The enzyme almost completely preserved its activity for 1 hour in distilled water at 60 degrees C. The temperature maximum of the enzyme activity was 70 degrees at pH 8. The enzyme may be referred to proteases of serine nature, because it is completely inactivated with diisopropylphosphofluoridate, but it retains the activity in the presence of chelating agents (EDTA, o-phenantroline, ditizone) and inhibitors of SH-groups (sodium p-chloromercuriumbenzoate, iodoacetic acid). The enzyme was not inactivated with phenylmethylsulphonylfluoride and the trypsin inhibitor from soybean. The protease studied most efficiently hydrolyzed caseine and hemoglobin, in a less degree--human serum albumin and fibrinogen and almost did not attack egg albumin. The enzyme undergoes association-dissociation under pH change during gel filtration through Sephadex.
[Partial purification and properties of protease from Torula thermophila]. 11-Fold purified protease preparation is isolated from cultural medium of Torula thermophila UzPT-1 by means of ammonium sulphate precipitation and gel chromatography through Sephadex G-100. Disc polyacrylamide gel electrophoresis revealed two portease components, one of them possessing proteolytic activity. pH interval for protease activity was found to be 3.5-12, the maximal activity was observed at pH 8.5-11, the highest enzyme resistance--at pH 6-8. The enzyme almost completely preserved its activity for 1 hour in distilled water at 60 degrees C. The temperature maximum of the enzyme activity was 70 degrees at pH 8. The enzyme may be referred to proteases of serine nature, because it is completely inactivated with diisopropylphosphofluoridate, but it retains the activity in the presence of chelating agents (EDTA, o-phenantroline, ditizone) and inhibitors of SH-groups (sodium p-chloromercuriumbenzoate, iodoacetic acid). The enzyme was not inactivated with phenylmethylsulphonylfluoride and the trypsin inhibitor from soybean. The protease studied most efficiently hydrolyzed caseine and hemoglobin, in a less degree--human serum albumin and fibrinogen and almost did not attack egg albumin. The enzyme undergoes association-dissociation under pH change during gel filtration through Sephadex.