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Solvent and thermal stability, and pH kinetics, of proline-specific dipeptidyl peptidase IV-like enzyme from bovine serum

Paper ID Volume ID Publish Year Pages File Format Full-Text
18726 42738 2007 5 PDF Available
Title
Solvent and thermal stability, and pH kinetics, of proline-specific dipeptidyl peptidase IV-like enzyme from bovine serum
Abstract

Proline-specific dipeptidyl peptidase-like (DPP IV; EC 3.4.14.5) activity in bovine serum has attracted little attention despite its ready availability and the paucity of useful proline-cleaving enzymes. Bovine serum DPP IV-like peptidase is very tolerant of organic solvents, particularly acetonitrile: upon incubation for 1 h at room temperature in 70% acetonitrile, 47% dimethylformamide, 54% DMSO and 33% tetrahydrofuran (v/v concentrations) followed by dilution into the standard assay mixture, the enzyme retained half of its aqueous activity. As for thermal performance in aqueous buffer, its relative activity increased up to 50 °C. Upon thermoinactivation at 71 °C, pH 8.0 (samples removed periodically, cooled on ice, then assayed under optimal conditions), residual activities over short times fit a first-order decay with a k-value of 0.071 ± 0.0034 min−1. Over longer times, residual activities fit to a double exponential decay with k1 and k2 values of 0.218 ± 0.025 min−1 (46 ± 4% of overall decay) and 0.040 ± 0.002 min−1 (54 ± 4% of overall decay), respectively.The enzyme's solvent and thermal tolerances suggest that it may have potential for use as a biocatalyst in industry. Kinetic analysis with the fluorogenic substrate Gly-Pro-7-aminomethylcoumarin over a range of pH values indicated two pK values at 6.18 ± 0.07 and at 9.70 ± 0.50. We ascribe the lower value to the active site histidine; the higher may be due to the active site serine or to a free amino group in the substrate.

Keywords
ACN, acetonitrile; ADAbp, adenosine deaminase binding protein; AMC, 7-amino-4-methylcoumarin; BCA, bicinchoninic acid; CD26, cluster of differentiation molecule 26; CHES, 2-(cyclohexylamino)ethanesulfonic acid; DMF, dimethylformamide; DMSO, dimethylsulfox
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Solvent and thermal stability, and pH kinetics, of proline-specific dipeptidyl peptidase IV-like enzyme from bovine serum
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 41, Issue 3, 2 August 2007, Pages 307–311
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