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Hindi Research Article
Purification, Determination Molecular Weight and Study Kinetic Properties of G6PD from Diabetes Patient
Abdulkader Rasheed W1, Firas Maher T1 and Akeel Al-Aisse H2*
1Department of Chemistry, College of Science, University of Tikrit, Tikrit, Iraq
2Department of Biology, College of Science, University of Tikrit, Tikrit, Iraq
- *Corresponding Author:
- Akeel Al-Aisse H
Department of Chemistry
College of Science, University
of Tikrit, Tikrit, Iraq
Tel: 9641543416
E-mail: akeel@yahoo.com
Received Date: June 07, 2017 Accepted Date: July 20, 2017 Published Date: July 25, 2017
Citation: Rasheed WA, Maher TF, Al-Aisse HA (2017) Purification, Determination Molecular Weight and Study Kinetic Properties of G6PD from Diabetes Patient. J Anal Bioanal Tech 8: 373. doi: 10.4172/2155-9872.1000373
Copyright: © 2017 Rasheed WA, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
This study was conducted to purification G6PD enzyme from diabetic patients by using simple and cheap method the technique gel filtration on Sephadex G100 and determine molecular weight of enzyme and compare it with true molecular weight of enzyme and determine kinetic constant (Km, Vmax) and study the effect of temperature and substrate and pH and known the best condition to give optimum work of enzyme. study contain (60) patients with diabetes and (60) control Glucose and activity of G6PD were measured and the enzyme precipitated by Ammonium Sulfate with concentration (75%) and purification enzyme gel filtration on Sephadex G-100 with dimensions (1.5 × 30) cm and using the buffer solution from (Tris-HCl) at pH 8.2 to isolate the enzyme and determine molecular weight with same method. Specific activity was calculated (21.5 UI/mg), total activity (706.8 UI), number of purification (3.45) enzyme yield (23.188%) and enzyme activity (17.67 UI/ml). and the molecular weight was calculated with using same technique (57.82) kD. Effect of increased concentration of substrate on enzyme activity and found the activity increase with increase substrate and amount constant level not change however increase of concentration of substrate when drawing relation between activity and concentration of substrate format appear exchange excess and after study effect of pH found the optimum value (8.4) and study effect of temperature on activity found (38 C) the optimum temperature. Study of kinetic constant was done and the and the Michaelis-Menten (Km) value was (3.8 mM) and Vmax value (8 IU/ml).
