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Purification and characterization of two types of alkaline serine proteases produced by an alkalophilic actinomycete
Author(s) -
Tsujibo Hiroshi,
Miyamoto Katsushiro,
Hasegawa Toru,
Inamori Yoshihiko
Publication year - 1990
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.1365-2672.1990.tb01544.x
Subject(s) - proteases , protease , sephadex , chemistry , serine protease , enzyme , biochemistry , chromatography , polyacrylamide gel electrophoresis , serine
T sujibo , H., M iyamoto , K., H asegawa , T. & I namori , Y. 1990. Purification and characterization of two types of alkaline serine proteases produced by an alkalophilic actinomycete. Journal of Applied Bacteriology 69 , 520–529. Two types of alkaline serine proteases were isolated from the culture filtrate of an alkalophilic actinomycete, Nocardiopsis dassonvillei OPC‐210. The enzymes (protease I and protease II) were purified by acetone precipitation, DEAE‐Sephadex A‐50, CM‐Sepharose CL‐6B, Sephadex G‐75 and phenyl‐Toyopearl 650 M column chromatography. The purified enzymes showed a single band on sodium dodecyl sulphate polyacrylamide gel electrophoresis. The molecular weights of proteases I and II were 21000 and 36000, respectively. The pIs were 6.4 (protease I) and 3.8 (protease II). The optimum pH levels for the activity of two proteases were pH 10–12 (protease I) and pH 10.5 (protease II). The optimum temperature for the activity of protease I was 70°C and that for protease II was 60°C. Protease I was stable in the range of pH 4.0–8.0 up to 60°C and protease II was stable in the range of pH 6.0–12.0 up to 50°C.