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Volume 23 |
April - June 2007 (Part-I) |
Number 2 |
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Published by : PROFESSIONAL MEDICAL PUBLICATIONS |
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ORIGINAL ARTICLE |
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Volume 23 |
April - June 2007 (Part-I) |
Number 2 |
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Characterization of an intracellular protease
from Pseudomonas Aeruginosa
Shahanara Begum1, Iftikhar Ahmed2, Faisal Alam3, Samsuzzaman4,
Parvez Hassan5, Nurul Absar6, Jalaluddin Ashraful Haq7
ABSTRACT
An intracellular protease was extracted and purified from Pseudomonas aeruginosa by ion-exchange chromatography on DEAE-cellulose followed by CM–cellulose and rechromatography on DEAE-cellulose. The purified protease was found to be homogeneous as judged by polyacrylamide disc gel electrophoresis (PAGE). The molecular mass of the protease as determined by gel filtration on G-150 was about 48,000 and about 49,000 on SDS-PAGE. The enzyme is monomeric in nature. The purified protease is a glycoprotein with neutral sugar content of 0.6%. The Km value of the protease was found to be 0.48% against casein as substrate. The enzyme is stable up to 600C and showed maximum activity around 500C. The enzyme activity was affected with the changes of pH and the maximum proteolytic activity was observed at pH 8.0. The protease activity was inhibited in the presence of EDTA, Cu2+, Mn2+and Hg2+ whereas the presence of Ca2+, K+, Na+ and ascorbic acid enhanced the activity.
KEY WORDS: Pseudomonas aeruginosa, Intracellular protease, Purification andCharacterization.
Pak J Med Sci April 2007 Vol. 23 No. 2 227-232
1. Dr. Shahanara Begum
Assistant Professor
2. Dr. Iftikhar Ahmed
Professor
3. Dr. Faisal Alam
Lecturer
1-3: Department of Microbiology,
Rajshahi Medical College, Rajshahi, Bangladesh.
4. Dr. Samsuzzaman
Department of Biochemistry and Molecular Biology
5. Dr. Parvez Hassan
Associate Professor, Institute of Biological Sciences
6. Dr. Nurul Absar
Department of Biochemistry and Molecular Biology
4-6: University of Rajshahi, Bangladesh.
7. Dr. Jalaluddin Ashraful Haq,
Department of Microbiology, BIRDEM, Dhaka, Bangladesh
Correspondence:
Dr. Shahanara Begum
E-Mail: faisal_imm@yahoo.com
* Received for Publication: April 4, 2006
* Revision Received: May 15, 2006
* Revision Accepted: September 30, 2006
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