通过荧光光谱滴定法测定牛α-糜蛋白酶、胰蛋白酶、凝血酶和Xa因子溶液的操作摩尔浓度。
Determination of the operational molarity of solutions of bovine alpha-chymotrypsin, trypsin, thrombin and factor Xa by spectrofluorimetric titration.
作者信息
Jameson G W, Roberts D V, Adams R W, Kyle W S, Elmore D T
出版信息
Biochem J. 1973 Jan;131(1):107-17. doi: 10.1042/bj1310107.
Abstract
Several esters of 4-methylumbelliferone and 2-naphthol were synthesized and examined as possible spectrofluorimetric titrants for bovine alpha-chymotrypsin, trypsin, thrombin, Factor Xa and for subtilisin Novo. 4-Methylumbelliferyl p-guanidinobenzoate hydrochloride (MUGB) is a satisfactory titrant for alpha- and beta-trypsin, thrombin and Factor Xa and 4-methylumbelliferyl p-(NNN-trimethylammonium)cinnamate (MUTMAC) is a good titrant for alpha-chymotrypsin. The amount of enzyme used for spectrofluorimetric titration is 0.02-3.00nmol and the amount of 4-methylumbelliferone liberated is independent of the concentration of titrant and stoicheiometrically equal to the amount of enzyme used. Results obtained with MUGB and MUTMAC have been checked by spectrophotometric titration with p'-nitrophenyl p-guanidinobenzoate hydrochloride and p-nitrophenyl N(2)-acetyl-N(1)-benzylcarbazate respectively. p-Nitrophenyl N(2)-acetyl-N(1)-(9-anthrylmethyl)carbazate has been synthesized; it did not react with alpha-chymotrypsin. A satisfactory spectrofluorimetric titrant for subtilisin Novo was not discovered.
摘要
合成了4-甲基伞形酮和2-萘酚的几种酯，并将其作为牛α-胰凝乳蛋白酶、胰蛋白酶、凝血酶、因子Xa和诺维信枯草杆菌蛋白酶的潜在荧光分光光度滴定剂进行了研究。4-甲基伞形酮对胍基苯甲酸盐酸盐（MUGB）是α-和β-胰蛋白酶、凝血酶和因子Xa的满意滴定剂，4-甲基伞形酮对（NNN-三甲基铵）肉桂酸盐（MUTMAC）是α-胰凝乳蛋白酶的良好滴定剂。用于荧光分光光度滴定的酶量为0.02 - 3.00nmol，释放的4-甲基伞形酮量与滴定剂浓度无关，化学计量上等于所用酶的量。用MUGB和MUTMAC得到的结果分别用对硝基苯基对胍基苯甲酸盐酸盐和对硝基苯基N(2)-乙酰基-N(1)-苄基氨基甲酸酯进行分光光度滴定进行了验证。合成了对硝基苯基N(2)-乙酰基-N(1)-(9-蒽甲基)氨基甲酸酯；它不与α-胰凝乳蛋白酶反应。未发现诺维信枯草杆菌蛋白酶的满意荧光分光光度滴定剂。
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p-Nitrophenyl-p'-guanidinobenzoate HCl: a new active site titrant for trypsin.对硝基苯基-对'-胍基苯甲酸盐酸盐：一种用于胰蛋白酶的新型活性位点滴定剂。
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The determination of the concentration of hydrolytic enzyme solutions: alpha-chymotrypsin, trypsin, papain, elastase, subtilisin, and acetylcholinesterase.水解酶溶液浓度的测定：α-胰凝乳蛋白酶、胰蛋白酶、木瓜蛋白酶、弹性蛋白酶、枯草杆菌蛋白酶和乙酰胆碱酯酶。
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Phenanthrenequinone as an analytical reagent for arginine and other monosubstituted guanidines.菲醌作为精氨酸及其他单取代胍的分析试剂。
Biochim Biophys Acta. 1966 Dec 28;130(2):538-40. doi: 10.1016/0304-4165(66)90256-x.
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A titration method for bovine trypsin.一种用于牛胰蛋白酶的滴定方法。
Anal Biochem. 1969 May;29(2):305-10. doi: 10.1016/0003-2697(69)90314-5.
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