Isolation, purification and characterization of Phospholipase D from almond (Amygdalus spinosissima) from Ziarat, Balochistan

This study was carried out for the isolation, purification, and characterization of enzyme phospholipase D (PLD) from Amygdalus spinosissima Bunge (wild thorny almond). It was observed that the crude extract showed higher values of enzyme activity 1206 U than that of purified which was found to be 9...

Full description

Saved in:
Bibliographic Details
Published in:Pure and applied biology 2022-06, Vol.11 (2), p.661-669
Main Authors: Baloch, Mahgul, Sajjad, Ashif, Rizwan, Sabeena, Anjum, Shaista, Akbar, Nukhba, Hafeez, Irfan, Mahmood, Khalid
Format: Article
Language:English
Subjects:
Alzheimer's disease
Amygdalus spinosissima
Calcium
Calcium ions
Chromatography
Electrophoresis
Enzymatic activity
Enzyme activity
Enzymes
Gel electrophoresis
Glycerol
Hydrophobicity
Industrial applications
Molecular weight
Neurodegeneration
Phospholipase
Phospholipase D
Proteins
Purification
Sodium dodecyl sulfate
Sodium lauryl sulfate
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:This study was carried out for the isolation, purification, and characterization of enzyme phospholipase D (PLD) from Amygdalus spinosissima Bunge (wild thorny almond). It was observed that the crude extract showed higher values of enzyme activity 1206 U than that of purified which was found to be 915 U. The optimum time for extraction was 1.5 hours, after which the activity started decreasing. Purification of enzyme was done by hydrophobic interaction chromatography with a specific effect of calcium using Octyl Sepharose. In the absence of calcium ion, the fraction eluted from the column showed high enzymatic activity. The highest specific activity (4575 Umg-1), purification fold (53.2) and yield (75.8%) were obtained. Molecular weight was estimated as 62 kDa by SDS-PAGE (sodium dodecyl sulfatepolyacrylamide gel electrophoresis) analysis. Highest activity of PLD was observed at 50°C and at 6 pH. Calcium ion stability of PLD activity was found at 40-90 mM. It is concluded from the present study that PLD has numerous industrial applications. It can be extracted and purified from A. spinosissima which is a cheap and easily available source at industrial scale.
ISSN:2304-2478
DOI:10.19045/bsoab.2022.110067