Studies on alkaline serine protease produced by Bacillus clausii GMBE 22

Prep Biochem Biotechnol. 2009;39(3):289-307. doi: 10.1080/10826060902953269.

Abstract

An alkali tolerant Bacillus strain having extracellular serine alkaline protease activity was newly isolated from compost and identified as Bacillus clausii GMBE 22. An alkaline protease (AP22) was 4.66-fold purified in 51.5% yield from Bacillus clausii GMBE 22 by ethanol precipitation and DEAE-cellulose anion exchange chromatography. The purified enzyme was identified as serine protease by LC-ESI-MS analysis. Its complete inhibition by phenylmethanesulfonylfluoride (PMSF) also justified that it is a serine alkaline protease. The molecular weight of the enzyme is 25.4 kDa. Optimal temperature and pH values are 60 degrees C and 12.0, respectively. The enzyme showed highest specificity to N-Suc-Ala-Ala-Pro-Phe-pNA. The K(m) and k(cat) values for hydrolysis of this substrate are 0.347 mM and 1141 min(-1) respectively. The enzyme was affected by surface active agents to varying extents. The enzyme is stable for 2 h at 30 degrees C and pH 10.5. AP22 is also stable for 5 days over the pH range 9.0-11.0 at room temperature. AP22 has good pH stability compared with the alkaline proteases belonging to other strains of Bacillus clausii reported in the literature.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacillus / classification
  • Bacillus / enzymology*
  • Base Sequence
  • Cations, Divalent / chemistry
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Stability
  • Hydrogen Peroxide / chemistry
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Molecular Weight
  • Phylogeny
  • Serine Endopeptidases / chemistry
  • Serine Endopeptidases / metabolism*
  • Substrate Specificity
  • Surface-Active Agents / chemistry
  • Temperature

Substances

  • Cations, Divalent
  • DNA Primers
  • Surface-Active Agents
  • Hydrogen Peroxide
  • Serine Endopeptidases