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Purification and biochemical characterization of a chymotrypsin-like serine protease from Euphorbia neriifolia Linn.

Paper ID Volume ID Publish Year Pages File Format Full-Text
35262 45083 2011 9 PDF Available
Title
Purification and biochemical characterization of a chymotrypsin-like serine protease from Euphorbia neriifolia Linn.
Abstract

Neriifolin, a chymotrypsin-like serine protease, has been purified from the latex of Euphorbia neriifolia Linn. by ammonium sulfate precipitation, cation exchange chromatography and gel filtration. The molecular mass of the enzyme is 35.24 kDa, with an isoelectric point of pH 5.7. The enzyme consists of 18 tryptophan, 25 tyrosine and 9 cysteine residues with 4 disulfide bridges. The extinction coefficient (ε280 nm1%) is 38.28. The Km values are 1.39 ± 0.08 mM and 1.94 ± 0.17 mM, with N-succinyl-l-Phe-p-nitroanilide and α-leucine-p-nitroanilide as substrates, respectively. Neriifolin retains proteolytic activity over a wide range of pH and temperature value, with pH optima of 8.5 and an optimal temperature of 55 °C. Inhibition of enzyme activity by chymostatin and amidolytic activity against synthetic substrates specific to chymotrypsin indicates that the enzyme belongs to chymotrypsin-like serine protease class. Polyclonal antibodies specific to neriifolin and immunodiffusion reveal that the enzyme has unique antigenic determinants. The amino terminal sequence of the first 14 residues of neriifolin is D–F–P–P–N–T–H–I–G–I–P–N–G–Y. A high ratio of milk-clotting activity to proteolytic activity as well as stability against variations in pH and temperature, surfactants, oxidizing agents and compatibility with detergent additives make neriifolin an excellent candidate for industrial applications.

Keywords
BLAST, basic local alignment search tool; BAPA, NR-benzoylarginine-p-nitroanilide; BSA, bovine serum albumin; DFP, diisopropylfluorophosphate; DMSO, dimethyl sulfoxide; DTNB, 5,5 μ-dithiobis (2-nitrobenzoic acid); DTT, dithiothreitol; EDTA, ethylenediamin
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Purification and biochemical characterization of a chymotrypsin-like serine protease from Euphorbia neriifolia Linn.
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 46, Issue 8, August 2011, Pages 1654–1662
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us