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Cold-active serine alkaline protease from the psychrophilic bacterium Pseudomonas strain DY-A: enzyme purification and characterization
- Source :
- Extremophiles. 7:335-337
- Publication Year :
- 2003
- Publisher :
- Springer Science and Business Media LLC, 2003.
-
Abstract
- An extracellular protease was purified from a deep-sea psychrophilic bacterium strain DY-A which was identified as a Pseudomonas species. The optimal growth and protease-producing temperatures of the strain were all 10 degrees C, and the protease was secreted only at temperatures under 20 degrees C. The enzyme was most active at 40 degrees C and at pH 10.0. It was inhibited by phenylmethyl sulfonylfluoride and diisopropyl fluorophosphate, indicating that it is a serine protease. Chelators such as EDTA, EGTA, 1,10-phenanthroline and 2,2'-bipyridyl produced a decrease of activity. The enzyme was sensitive to denaturing agents such as SDS, urea, and guanidine HCl and resistant to thiol-containing reducing agents such as dithiotreitol. The enzyme was active towards N-succinyl-Ala-Ala-Pro-Phe- p-nitroanilide and N-succinyl-Ala-Ala-Pro-Leu- p-nitroanilide. The native molecular mass of the enzyme determined by native PAGE and SDS-PAGE was 25 kDa.
- Subjects :
- Protein Denaturation
Serine Proteinase Inhibitors
medicine.medical_treatment
Microbiology
Chromatography, DEAE-Cellulose
Substrate Specificity
Serine
Pseudomonas
Enzyme Stability
medicine
Psychrophile
chemistry.chemical_classification
Serine protease
Protease
biology
Molecular mass
Serine Endopeptidases
Temperature
General Medicine
Hydrogen-Ion Concentration
biology.organism_classification
Cold Temperature
Enzyme
chemistry
Biochemistry
biology.protein
Molecular Medicine
Diisopropyl fluorophosphate
Electrophoresis, Polyacrylamide Gel
medicine.drug
Subjects
Details
- ISSN :
- 14334909 and 14310651
- Volume :
- 7
- Database :
- OpenAIRE
- Journal :
- Extremophiles
- Accession number :
- edsair.doi.dedup.....247a0c6821a513742a3a273d0ab08df8