Kochmann G, Kochmann R, Pape W, Blank M, Baumann K
J Clin Chem Clin Biochem. 1983 Aug;21(8):503-9. doi: 10.1515/cclm.1983.21.8.503.
Most biological fluids contain both neutral and acid alpha-glucosidase. Optimal conditions were therefore developed for the selective determination of the activity of neutral and acid alpha-glucosidase, using 2-step, discontinuous assays. In the first step of the assay of neutral alpha-glucosidase, glucose was liberated from maltose (citrate-phosphate buffer, pH 6.8, 20 mmol/l maltose, 25 mmol/l turanose). Under these incubation conditions, turanose inhibited the residual activity of acid alpha-glucosidase almost completely without influencing the activity of neutral alpha-glucosidase. In the first step of the acid alpha-glucosidase assay, glucose was liberated from maltose (citrate-phosphate buffer, pH 3.8, 50 mmol/l maltose, 2 mol/l potassium chloride). Under these incubation conditions, potassium ions stimulate the activity of acid alpha-glucosidase and simultaneously inhibit almost completely the residual activity of neutral alpha-glucosidase. In the second step of the assay of neutral and acid alpha-glucosidase, the liberated glucose was measured by hexokinase/glucose-6-phosphate dehydrogenase. The effect of turanose and potassium ions on neutral and acid alpha-glucosidase from human urine was characterized.
大多数生物体液中都含有中性和酸性α-葡萄糖苷酶。因此，采用两步、不连续测定法，开发了用于选择性测定中性和酸性α-葡萄糖苷酶活性的最佳条件。在中性α-葡萄糖苷酶测定的第一步中，葡萄糖从麦芽糖中释放出来（柠檬酸-磷酸盐缓冲液，pH 6.8，20 mmol/L麦芽糖，25 mmol/L松二糖）。在这些孵育条件下，松二糖几乎完全抑制酸性α-葡萄糖苷酶的残留活性，而不影响中性α-葡萄糖苷酶的活性。在酸性α-葡萄糖苷酶测定的第一步中，葡萄糖从麦芽糖中释放出来（柠檬酸-磷酸盐缓冲液，pH 3.8，50 mmol/L麦芽糖，2 mol/L氯化钾）。在这些孵育条件下，钾离子刺激酸性α-葡萄糖苷酶的活性，同时几乎完全抑制中性α-葡萄糖苷酶的残留活性。在中性和酸性α-葡萄糖苷酶测定的第二步中，通过己糖激酶/葡萄糖-6-磷酸脱氢酶测量释放的葡萄糖。对松二糖和钾离子对人尿中中性和酸性α-葡萄糖苷酶的影响进行了表征。
