SPECIFIC ENZYMATIC ASSAY FOR D-GLUCARATE IN HUMAN SERUM
SPECIFIC ENZYMATIC ASSAY FOR D-GLUCARATE IN HUMAN SERUM
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DOI：
10.1016/0003-2697(90)90294-j
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发表时间：
1990-03-01
影响因子：
2.9
通讯作者：
BLUMENTHAL, DC
中科院分区：
文献类型：
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作者：
BLUMENTHAL, HJ;LUCUTA, VL;BLUMENTHAL, DC
建立了一种灵敏、特异的分光光度法测定人血清中d -葡萄糖酸盐水平的方法。该试验利用大肠杆菌葡萄糖酸分解代谢酶d -葡萄糖酸脱氢酶，α -酮- β。-脱氧d -葡二酸醛缩酶和酒石酸半醛还原酶，将d -葡二酸转化为等量的丙酮酸和TSA。在包括NADH，乳酸脱氢酶和三种大肠杆菌酶的单管反应中，1。D-glucarate定量转化为1 .mu。d -甘油三酯和l -乳酸各mol，同时利用2 μ m。mol NADH。使用该方法，测定血清中的d -葡萄糖酸盐，并从添加了d -葡萄糖酸盐的重复血清样品中定量回收真实的d -葡萄糖酸盐。葡萄糖酸盐是一种主要的血清有机酸，近似于以前由其他人测定的血丙酮酸水平。
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 .mu.mol of D-glucarate was quantitatively converted to 1 .mu.mol each of D-glycerate and L-lactate with concomitant utilization of 2 .mu.mol 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.