Studies on Alkaline Serine Protease Produced byBacillus clausiiGMBE 22
Autor: | Hasan Umit Ozturk, Dilek Coşkuner Öztürk, Altan Erarslan, Hulya Bal, Aydan Salman Dilgimen, Dilek Kazan, Aziz Akin Denizci, Nurcin Celik Ozturk |
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Rok vydání: | 2009 |
Předmět: |
Proteases
Cations Divalent Molecular Sequence Data Bacillus Biochemistry Substrate Specificity Serine Surface-Active Agents Hydrolysis chemistry.chemical_compound Enzyme Stability Amino Acid Sequence Phylogeny DNA Primers Serine protease chemistry.chemical_classification Chromatography Base Sequence biology Serine Endopeptidases Bacillus clausii Temperature Substrate (chemistry) Hydrogen Peroxide General Medicine Hydrogen-Ion Concentration biology.organism_classification Molecular Weight Enzyme chemistry biology.protein Electrophoresis Polyacrylamide Gel PMSF Biotechnology |
Zdroj: | Preparative Biochemistry and Biotechnology. 39:289-307 |
ISSN: | 1532-2297 1082-6068 |
DOI: | 10.1080/10826060902953269 |
Popis: | 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. |
Databáze: | OpenAIRE |
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