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Research Article
A Protease Isolated from the Latex of Plumeria rubra
Linn (Apocynaceae) 1: Purification and
Characterization
Indranil Chanda1*,
Sanat Kumar Basu2, Sadhan Kumar Dutta3
and Smriti Rekha Chanda Das1
1Girijananda
Chowdhury Institute of Pharmaceutical Science, Guwahati,
Assam-781017, 2Department of Pharmaceutical
Technology, Jadavpur University, Kolkata, West
Bengal-700032, 3A College of Pharmacy, Bengal
School of Technology, Hooghly, West Bengal- 712102,
India.
For correspondence:
E-mail:
ichanda@sify.com
Tel: +91-9957179226
Received: 12 May
2011 Revised
accepted: 15 October, 2011
Tropical
Journal of Pharmaceutical Research, December 2011;
10(6): 705-711
http://dx.doi.org/10.4314/tjpr.v10i6.2
Abstract
Purpose:
To isolate, purify and characterize protease from the
latex of the plant.
Methods:
Protease was isolated from
the latex of Plumeria rubra Linn using acetone
precipitation method and purified by a sequence of DEAE
cellulose column chromatography, followed by two
successive column purification in Sephadex G-50 and
Sephadex G-200. The molecular weight of the purified
protease was determined by
sodium dodecyl sulfate
polyacrylamide gel electrophoresis (SDS-PAGE).
The protease was given a trivial name, Plumerin-R.
Results:
Plumerin-R showed a single protein band on SDS-PAGE and
molecular weight was approximately 81.85 kDa. It
remained active over a broad range of temperature but
had optimum activity at 55 °C and pH 7.0 when casein was
used as substrate. Activation of the protease by a thiol-activating
agent indicated the presence of sulfhydryl as an
essential group for its activity.
Conclusion:
A protease from the latex of Plumeria rubra Linn was
purified to homogeneity by a simple purification
procedure and then characterized.
Keywords:
Protease, Plumerin-R,
Sulfhydryl, Purification; Characterization |
