Purification and characterization of two types of alkaline serine proteases produced by an alkalophilic actinomycete

J Appl Bacteriol. 1990 Oct;69(4):520-9. doi: 10.1111/j.1365-2672.1990.tb01544.x.

Abstract

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 21,000 and 36,000, 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 temperture for the activity of protease I was 70 degrees C and that for protease II was 60 degrees C. Protease I was stable in the range of pH 4.0-8.0 up to 60 degrees C and protease II was stable in the range of pH 6.0-12.0 up to 50 degrees C.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actinomycetales / classification
  • Actinomycetales / enzymology*
  • Cations, Divalent / pharmacology
  • Chelating Agents / pharmacology
  • Electrophoresis, Polyacrylamide Gel
  • Hydrogen-Ion Concentration
  • Isoelectric Point
  • Molecular Weight
  • Serine Endopeptidases / isolation & purification*
  • Serine Endopeptidases / metabolism
  • Temperature

Substances

  • Cations, Divalent
  • Chelating Agents
  • Serine Endopeptidases
  • microbial serine proteinases