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Coupled-Enzyme System for Measuring Viral Neuraminidase Activity

机译：耦合酶系统测量病毒神经氨酸酶活性

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摘要

A coupled-enzyme assay for determining viral neuraminidase activity is described. All reactants—viral neuraminidase, the initial substrate (fetuin), N-acetylneuraminic acid aldolase, lactic acid dehydrogenase, and reduced nicotinamide adenine dinucleotide—are combined in a single cuvette. Thus, in a single coupled system neuraminidase releases N-acetylneuraminic acid, which is cleaved to N-acetyl-D-mannosamine and pyruvic acid; finally, pyruvate is reduced to lactate as reduced nicotinamide adenine dinucleotide is oxidized. The rate of change of absorbance at 340 nm, as reduced nicotinamide adenine dinucleotide is oxidized, is a measure of the rate of reaction of the coupled system. This procedure, which measures the rate of release of N-acetylneuraminic acid by neuraminidase, is an alternate method for those procedures which require multistep, colorimetric determinations.
机译：描述了用于确定病毒神经氨酸酶活性的偶联酶测定法。所有反应物（病毒神经氨酸酶，起始底物（胎蛋白），N-乙酰神经氨酸醛缩酶，乳酸脱氢酶和还原烟酰胺腺嘌呤二核苷酸）都合并在一个比色皿中。因此，在单个偶联的系统中，神经氨酸酶释放N-乙酰神经氨酸，其被裂解为N-乙酰-D-甘露糖胺和丙酮酸。最后，当还原的烟酰胺腺嘌呤二核苷酸被氧化时，丙酮酸被还原为乳酸。当还原的烟酰胺腺嘌呤二核苷酸被氧化时，在340 nm处吸光度的变化速率是偶联系统反应速率的量度。该方法测量神经氨酸酶释放N-乙酰神经氨酸的速率，是需要多步比色测定的那些方法的另一种方法。

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