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Cow Brain Glutaminase: Purification And Influence Of Phosphate And Borate Ions On Its Activity And Molecular Weight, Mahnaz Badamchian
Cow Brain Glutaminase: Purification And Influence Of Phosphate And Borate Ions On Its Activity And Molecular Weight, Mahnaz Badamchian
All Graduate Theses and Dissertations, Spring 1920 to Summer 2023
An efficient and simple purification procedure for cow brain glutaminase (L- glutamine amidohydrolase, EC 3.5.1.2) is described. The main steps consisted of acetone extraction and French press treatment in the presence of phosphate ions, precipitation of nucleic acids and lipids by 0.08 % protamine·S04 and high speed centrifugation (300,000xg), ammonium sulfate fractionation, and gel filtration on Sepharose 4B first as the low molecular weight and then as the aggregated form. The yield was 22% and the final preparation had a specific activity of 142 μmoles/min/mg. The purification was more than 8000-fold over crude brain homogenate. The enzyme showed one strong …