Volume : II, Issue : IV, April - 2013

Molecular Weight Determination of Glutaminase Enzyme Produced from Erwinia

Pushpinder Paul

Abstract :

The purpose of this study was to determine molecular weight of glutaminase enzyme by SDS-PAGE.Glutaminase enzyme was produced from Erwinia under optimized condition. After production of glutaminase enzyme it was isolated by acetone precipitation & ammonium sulphate precipitation method. After isolation purification of enzyme was done by Gel Filtration chromatography and Ion Exchange chromatography. Then SDS-PAGE (Sodium dodecyl sulphate polyacrylamide gel electrophoresis) was performed to determine the molecular weight and the purity of the sample. It was observed after SDS-PAGE, that the number of bands after acetone extraction were less as compared to number of bands observed for protein crude sample before acetone extraction. Also the number of bands observed after ammonium sulphate fractionation are less. So the ammonium sulphate fractions was subjected in Gel Filtration which showed further purification. The Ion exchange chromatography showed more diffusion and less recovery of protein sample as compared to Gel Filtration Chromatography. This was confirmed by the achievement of one band during SDS-PAGE which confirmed the purity of enzyme. From the results of SDS-PAGE ,it was clear that molecular weight of Glutaminase isolated from Erwinia species came to be 145 Kda.

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Article: Download PDF   DOI : 10.36106/ijsr  

Cite This Article:

Pushpinder Paul Molecular Weight Determination of Glutaminase Enzyme Produced from Erwinia International Journal of Scientific Research, Vol.II, Issue.IV April 2013

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