Phosphotransferase activity in acid phosphatase in camel heart

Two forms of low molecular weight acid phosphatase (lmw-acpase) from camel heart were purified by salt fractionation,ion exchange chromatography,gel filtration and affinity chromatography. these were designated as peak-1 and peak-2. the peak-1 was purified 500-fold with specific activity of 28 u/mg of protein and a yield of 3 %,while peak-2 was purified 850 times with specific activity of 44 u/mg. the recovery was 4 %. the homogeneity of the enzymes was checked on sds-polyacrylamide gel electrophoresis (sds-page). both forms showed similar patterns on sds-page and had molecular weight of 18 kda. the lmw-acpase activity was increased in the presence of alcohols. this activation reflected phosphotransferase activity. the presence of various alcohols in the incubation mixture which acted as phosphate acceptors increased the overall rate of breakdown of substrate p-nitrophenyl phosphate as judged by the amount of pnitrophenol released and the competition between hydrolysis and phosphate transfer reaction was started. the rate of transfer of phosphate group to phosphate acceptors was increased while the amount of inorganic phosphate (pi) produced remained same and was equal to the amount of pi released in water when no alcohol was added. as the alcohol concentrations increased,the rate of formation of p-nitrophenol and the rate of pi transfer were greatly increased while the level of inorganic phosphate produced was found constant. thus,smaller than 1 ratios of pi/p-nitrophenol and greater ratios of phosphate transfer to hydrolysis showed the presence of transphosphorylation reaction.