Purification and characterization of heat-tolerant protease produced by Bacillus polyfermenticus SCD

J Microbiol Biotechnol. 2013 Nov 28;23(11):1554-9. doi: 10.4014/jmb1306.06073.

Abstract

A protease produced by Bacillus polyfermenticus SCD was purified and characterized as a new detergent material. The protease was purified from supernatant produced by B. polyfermenticus SCD, by ammonium sulfate precipitation, ion-exchange chromatography on a DEAE-Sephadex A-50, and finally gel filtration chromatography on Sephadex G-50. The molecular mass of this enzyme was 44 kDa based on SDS-PAGE. The optimum temperature and pH were 50°C and pH 8.0. The ranges of its stability to the pH and temperature were 7.0 to 9.0 and under 40°C, respectively. The enzyme was highly stable in the presence of the surfactants like Triton X-100 (0.1%), showing a 2-fold increase in its proteolytic activity. However, the enzyme was slightly inhibited by the chelating agent EDTA (1 mM). The enzyme has a maximum activity at 50°C and the activity can be increased by surfactants such as Triton X-100 and Tween 80.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus / enzymology*
  • Chemical Precipitation
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Molecular Weight
  • Peptide Hydrolases / chemistry*
  • Peptide Hydrolases / isolation & purification*
  • Peptide Hydrolases / metabolism
  • Temperature

Substances

  • Peptide Hydrolases