人血清中D-葡萄糖醛酸的特异性酶法测定。

Specific enzymatic assay for D-glucarate in human serum.

作者信息

Blumenthal H J, Lucuta V L, Blumenthal D C

机构信息

Department of Microbiology, Loyola University of Chicago, Stritch School of Medicine, Maywood, Illinois 60153.

出版信息

Anal Biochem. 1990 Mar;185(2):286-93. doi: 10.1016/0003-2697(90)90294-j.

DOI:10.1016/0003-2697(90)90294-j

PMID:2187374

Abstract

A sensitive and specific spectrophotometric assay was developed to determine levels of D-glucarate in human serum. This assay makes use of the Escherichia coli glucarate catabolic enzymes D-glucarate dehydrase, alpha-keto-beta-deoxy-D-glucarate aldolase, and tartronate semialdehyde (TSA) reductase, to convert D-glucarate to equimolar quantities of pyruvate and TSA. In a one-tube reaction that included NADH, lactate dehydrogenase, and the three E. coli enzymes, 1 mumol of D-glucarate was quantitatively converted to 1 mumol each of D-glycerate and L-lactate with concomitant utilization of 2 mumol of NADH. Using this method, D-glucarate in serum was measured, along with quantitative recovery of authentic D-glucarate from duplicate serum samples to which it had been added. Glucarate is a major serum organic acid, approximating blood pyruvate levels previously determined by others.

摘要

开发了一种灵敏且特异的分光光度法来测定人血清中D-葡糖二酸的水平。该测定法利用大肠杆菌葡糖二酸分解代谢酶D-葡糖二酸脱水酶、α-酮-β-脱氧-D-葡糖二酸醛缩酶和酒石酸半醛（TSA）还原酶，将D-葡糖二酸转化为等摩尔量的丙酮酸和TSA。在一个包含NADH、乳酸脱氢酶和三种大肠杆菌酶的单管反应中，1 μmol的D-葡糖二酸被定量转化为各1 μmol的D-甘油酸和L-乳酸，同时消耗2 μmol的NADH。使用该方法测定了血清中的D-葡糖二酸，并从添加了纯D-葡糖二酸的重复血清样本中实现了其定量回收。葡糖二酸是一种主要的血清有机酸，其水平与其他人先前测定的血液丙酮酸水平相近。