Characterization and intermethod relationships of materials containing purified human pancreatic and salivary amylase.

Abstract

We describe the preparation and characterization of materials containing human pancreatic and salivary alpha-amylase (EC 3.2.1.1) and examine their relationship to endogenous amylase in human serum. Amylase was purified from human pancreas and saliva by solvent- and salt-fractionation and column chromatography to specific activities of 63 and 279 kU/g, respectively. Four liquid pools, differing only in activity, were prepared from each source of amylase, each in a matrix containing, per liter: 30 g of human albumin, 50 mmol of sodium chloride, 1 mmol of calcium chloride, and 50 mmol of Tris hydrochloride buffer, pH 7.4. Characterization of the pools showed that the amylase activity in the materials was stable for at least six months at 25 degrees C; among-vial variability of amylase activity was less than or equal to 0.5% (2 CV); and the pools were free from eight possible contaminating enzymes. Plots of salivary vs pancreatic amylase activity measure in our materials with eight commercially available methods showed least-squares slopes ranging from 0.51 to 1.0. The intermethod "commutability" of the materials (i.e., how closely they mimic endogenous serum amylase) was examined in relationship to approximately 100 human sera.

Inhibition, Characterization and Purification of α-AMY from Sera of Iraqi Breast Cancer Patients

“…According to the data, the average values of Km and Vmax of α-AMY were 0.714 mM and 0.047 mM/min, respectively. The inhibitors were no effect on Km, while Vmax in presence of 75 μg/ml of each of the extracts was decreased to 0.023, and 0.012 mmol min -l , respectively (25) .…”

“…Wasan and Firas, revealed that the specific activity of α-AMY enzyme purified from the blood of diabetic patients was 21.8 U/mg and the degree of purification reached 16.1 with an enzymatic outcome of 108.2% using Ion exchange chromatography and Sephadex G-100 gel filtration column chromatography (24) . From the human pancreas and saliva, α-AMY was purified using solvent and salt fractionation and column chromatography to achieve specific activities of 63 and 279 kU/g, respectively (25) . -----------------------------------------------------------------------------------------------------------------------------------Determination of α-AMY molecular weight By using gel filtration chromatography, the mol.wt.…”

Salivary Amylase as Potential Biochemical Marker in Diabetes Mellitus

“…3 Human salivary amylase (α1, 4 α-D glucan, 4 gluconohydrolase) was first described by Leuchs in the year 1831. 4 Human salivary amylase is a major component of human salivary secretions, responsible for hydrolysis of starch into small sugar molecules.…”