用于测量病毒神经氨酸酶活性的偶联酶系统。
Coupled-enzyme system for measuring viral neuraminidase activity.
作者信息
Ziegler D W, Hutchinson H D
出版信息
Appl Microbiol. 1972 Jun;23(6):1060-6. doi: 10.1128/am.23.6.1060-1066.1972.
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.
本文描述了一种用于测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、初始底物（胎球蛋白）、N-乙酰神经氨酸醛缩酶、乳酸脱氢酶和还原型烟酰胺腺嘌呤二核苷酸——都在一个比色皿中混合。因此，在一个单一的偶联系统中，神经氨酸酶释放出N-乙酰神经氨酸，后者被裂解为N-乙酰-D-甘露糖胺和丙酮酸；最后，随着还原型烟酰胺腺嘌呤二核苷酸被氧化，丙酮酸被还原为乳酸。在340nm处吸光度的变化速率，即还原型烟酰胺腺嘌呤二核苷酸被氧化时的速率，是偶联系统反应速率的一种度量。该方法通过测量神经氨酸酶释放N-乙酰神经氨酸的速率，为那些需要多步比色测定的方法提供了一种替代方法。
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