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Purification and characterization of a new thermoalkaliphilic pectate lyase from Actinomadura keratinilytica Cpt20

Paper ID Volume ID Publish Year Pages File Format Full-Text
34221 45010 2015 8 PDF Available
Title
Purification and characterization of a new thermoalkaliphilic pectate lyase from Actinomadura keratinilytica Cpt20
Abstract

•A new extracellular pectate lyase from A. keratinilytica Cpt20.•This pectate lyase was purified (Pel-20) and biochemically characterized.•Optimal pH and temperature for activity were 10.5 and 70 °C respectively.•The Km and Vmax values were 0.45 mM and 21,700 U/mg respectively.•Pel-20 with remarkable properties is suitable for industrial bioprocesses.

This study was carried out to investigate the purification and biochemical characterization of a new extracellular alkaliphilic and thermostable pectate lyase (Pel-20) isolated from Actinomadura keratinilytica strain Cpt20. Pure protein was obtained after sequential chromatographies on a fast performance liquid chromatography (FPLC) and high performance liquid chromatography (HPLC) columns. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis revealed that the purified enzyme was a monomer with a molecular mass of 34125.11-Da. The enzyme had an NH2-terminal sequence of GFATNQGGTTGGAGGTLS, thus, sharing high homology with actinomycetes pectate lyase family. The results showed that this enzyme was completely inhibited by EDTA, which supports its belonging to the pectate lyase superfamily. It showed optimum activity at pH 10.5 and 70 °C. The thermoactivity and thermostability of Pel-20 were enhanced in the presence of 1 mM Ca2+. Its half-life times at 70, 80, 90, and 100 °C were 18, 12, 7, and 2 h, respectively. Its kinetic parameters, Km and Vmax values were 0.45 mM and 21,700 U/mg, respectively. Low-esterified pectin was the optimum substrate for the Pel-20. However, higher-esterified pectin was also weakly cleaved. Overall, the alkaliphilicity and thermostability properties of Pel-20 make it a potential candidate for future application in industrial bioprocesses.

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Keywords
Thermophilic actinomycetes; Pectate lyase; Purification; MALDI-TOF/MS; Kinetic parameters
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Purification and characterization of a new thermoalkaliphilic pectate lyase from Actinomadura keratinilytica Cpt20
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 50, Issue 12, December 2015, Pages 2259–2266
Authors
, , , , , , , , , ,
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