Purification and biochemical characterization of alkaline serine protease from caesalpinia bonducella

A high molecular weight serine protease has been purified to electrophoretic homogeneity from the seeds of Caesalpinia bonducella Hem. (Caesalpiniaceae) by the combination of size exclusion and ion exchange chromatography. About 524 fold purification was achieved with an overall recovery of 6.8%. Th...

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Main Authors:	Khan, H., Ali, I., Khan, Arif-ullah, Ahmed, M., Shah, Z., Saeed, A., Naz, R., Mustafa, M.R., Abbasi, A.
Format:	Article
Published:	Natural Products Inc 2010
Subjects:	Q Science (General)
Online Access:	http://eprints.um.edu.my/14173/

id	um-14173
recordtype	eprints
spelling	um-141732015-10-06T07:06:11Z Purification and biochemical characterization of alkaline serine protease from caesalpinia bonducella Khan, H. Ali, I. Khan, Arif-ullah Ahmed, M. Shah, Z. Saeed, A. Naz, R. Mustafa, M.R. Abbasi, A. Q Science (General) A high molecular weight serine protease has been purified to electrophoretic homogeneity from the seeds of Caesalpinia bonducella Hem. (Caesalpiniaceae) by the combination of size exclusion and ion exchange chromatography. About 524 fold purification was achieved with an overall recovery of 6.8%. The specific activity was found to be 86 U/mg/min at pH 8.0. The calculated K(m) and V(max) were 1.66 mg/mL and 496.68 units/min per mg of protein, respectively. The molecular mass was estimated to be about 63 kDa by sodium dodecyl sulfate PAGE. The enzyme showed optimum activity at pH 8.0 and exhibited its highest activity at 40 degrees C. The enzyme was strongly inhibited by 2mM phenylmethylsulfonyl fluoride (PMSF), suggesting the presence of a serine residue at the active site. PMSF showed a pure competitive type of inhibition with the serine protease enzyme. It was observed that enzyme activity was enhanced in the presence of dications and was active against a variety of modified substrates and natural proteins. Natural Products Inc 2010 Article PeerReviewed Khan, H.; Ali, I.; Khan, Arif-ullah; Ahmed, M.; Shah, Z.; Saeed, A.; Naz, R.; Mustafa, M.R.; Abbasi, A. (2010) Purification and biochemical characterization of alkaline serine protease from caesalpinia bonducella. Natural Product Communications <http://eprints.um.edu.my/view/publication/Natural_Product_Communications.html>, 5 (6). pp. 931-934. http://eprints.um.edu.my/14173/
repository_type	Digital Repository
institution_category	Local University
institution	University Malaya
building	UM Research Repository
collection	Online Access
topic	Q Science (General)
spellingShingle	Q Science (General) Khan, H. Ali, I. Khan, Arif-ullah Ahmed, M. Shah, Z. Saeed, A. Naz, R. Mustafa, M.R. Abbasi, A. Purification and biochemical characterization of alkaline serine protease from caesalpinia bonducella
description	A high molecular weight serine protease has been purified to electrophoretic homogeneity from the seeds of Caesalpinia bonducella Hem. (Caesalpiniaceae) by the combination of size exclusion and ion exchange chromatography. About 524 fold purification was achieved with an overall recovery of 6.8%. The specific activity was found to be 86 U/mg/min at pH 8.0. The calculated K(m) and V(max) were 1.66 mg/mL and 496.68 units/min per mg of protein, respectively. The molecular mass was estimated to be about 63 kDa by sodium dodecyl sulfate PAGE. The enzyme showed optimum activity at pH 8.0 and exhibited its highest activity at 40 degrees C. The enzyme was strongly inhibited by 2mM phenylmethylsulfonyl fluoride (PMSF), suggesting the presence of a serine residue at the active site. PMSF showed a pure competitive type of inhibition with the serine protease enzyme. It was observed that enzyme activity was enhanced in the presence of dications and was active against a variety of modified substrates and natural proteins.
format	Article
author	Khan, H. Ali, I. Khan, Arif-ullah Ahmed, M. Shah, Z. Saeed, A. Naz, R. Mustafa, M.R. Abbasi, A.
author_facet	Khan, H. Ali, I. Khan, Arif-ullah Ahmed, M. Shah, Z. Saeed, A. Naz, R. Mustafa, M.R. Abbasi, A.
author_sort	Khan, H.
title	Purification and biochemical characterization of alkaline serine protease from caesalpinia bonducella
title_short	Purification and biochemical characterization of alkaline serine protease from caesalpinia bonducella
title_full	Purification and biochemical characterization of alkaline serine protease from caesalpinia bonducella
title_fullStr	Purification and biochemical characterization of alkaline serine protease from caesalpinia bonducella
title_full_unstemmed	Purification and biochemical characterization of alkaline serine protease from caesalpinia bonducella
title_sort	purification and biochemical characterization of alkaline serine protease from caesalpinia bonducella
publisher	Natural Products Inc
publishDate	2010
url	http://eprints.um.edu.my/14173/
first_indexed	2018-09-06T06:20:40Z
last_indexed	2018-09-06T06:20:40Z
_version_	1610838012643508224

Purification and biochemical characterization of alkaline serine protease from caesalpinia bonducella

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