Leila Parsiavash; Azra Saboora; Seyedeh Zahra Moosavi Nejad
Abstract
In the present study, some techniques were used for Soybean peroxidase (SBP) purification including: ammonium sulfate fractionation, DEAE Sephadex anion exchange chromatography and Concanavalin A Sepharose 4B affinity chromatography. Molecular weight of purified SBP was estimated about 44 kDa by SDS-PAGE ...
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In the present study, some techniques were used for Soybean peroxidase (SBP) purification including: ammonium sulfate fractionation, DEAE Sephadex anion exchange chromatography and Concanavalin A Sepharose 4B affinity chromatography. Molecular weight of purified SBP was estimated about 44 kDa by SDS-PAGE as a single polypeptide band. The optimal pH and temperature for enzyme activity were found to be 4.5 and 70◦C, respectively. The enzyme was more stable in alkaline pH than acidic ones and could tolerate 10 minutes heating in 40-50◦C without any loss of its activity. Both NaCl and KCl were found to have significant effects on the enzyme stability, but presence of NaCl was more effective than KCl. Our results showed that after 24 hours incubation of the enzyme in the presence of 20 mM NaCl, more than 60٪ of the enzyme activity was remained while it would fall to 3٪ if incubation was not accompanied by NaCl. Purified peroxidase from seed hull of soybean relative to the other identified peroxidases was more stable, for this reason a lot of benefit will be considered by use of this enzyme in different industry.
Mehran Miroliaei; Samira Padidar; Ali Mostafaie; Sirous Ghobadi
Abstract
Abstract
Plant nonspecific lipid transfer proteins (nsLTPs) are divided into nsLTP1 and nsLTP2. The existence of an
internal hydrophobic cavity, is a typical characteristic of nsLTPs that serves as the binding site for lipid
substrates. In this communication a simple, rapid and low-cost alternative ...
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Abstract
Plant nonspecific lipid transfer proteins (nsLTPs) are divided into nsLTP1 and nsLTP2. The existence of an
internal hydrophobic cavity, is a typical characteristic of nsLTPs that serves as the binding site for lipid
substrates. In this communication a simple, rapid and low-cost alternative method was developed for
purification of nsLTP2 from rice paddy. After extracting, final supernatant was loaded on CM-Sepharose
column, which had previously equilibrated with 0.05 M Tris-HCl buffer, pH 8. Bounded proteins were
separated using a linear gradient of 0-0.5 M NaCl. Solution of separated proteins was dialyzed and applied on a
Phenyl-Sepharose column which previously equilibrated with Tris-HCl 0.05 M, ammonium sulfate 1.5 M,
EDTA 0.005 M and NaHSO3 0.3%, pH 8.4. Tris-Tricin SDS-PAGE of separated proteins, obtained from ionexchange
column, showed multiple bands in the range of 2-20 kDa. Further purification using hydrophobic
column resulted in single band of nsLTP2 at about 7 kDa, reflecting a purified sample in the gel.