Synthesis and Characterization of Luciferin Derivatives for Use in Bioluminescence Enhanced Enzyme Immunoassays. New Ultrasensitive Detection Systems for Enzyme Immunoassays, I.
Synthesis and Characterization of Luciferin Derivatives for Use in Bioluminescence Enhanced Enzyme Immunoassays. New Ultrasensitive Detection Systems for Enzyme Immunoassays, I.
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用于生物发光增强酶免疫测定的荧光素衍生物的合成和表征。
DOI：
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发表时间：
1987
期刊：
影响因子：
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通讯作者：
R. Geiger
中科院分区：
文献类型：
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作者：
W. Miska;R. Geiger
合成了作为高灵敏度底物用于酶测定的D-甜菜碱甲酯、D-甜菜碱-L-苯丙氨酸、D-甜菜碱-L-N α-精氨酸、D-甜菜碱-O-硫酸盐和D-甜菜碱-O-磷酸盐的甜菜碱衍生物。通过紫外分光光度法、荧光分光光度法、氨基酸分析法和快原子轰击质谱法对所合成的化合物进行了表征。证明了酶对化合物的酶裂解导致D-胡萝卜素的释放。测定了以下酶/底物对的动力学常数：D-精氨酸甲酯/羧酸酯酶、D-乙酰-L-苯丙氨酸/羧肽酶A、D-乙酰-L-N α-精氨酸/羧肽酶B、D-乙酰-O-硫酸酯/芳基硫酸酯酶、D-乙酰-O-磷酸酯/碱性磷酸酶。所有的化合物被证明是可接受的底物的各自的酶，D-甜菜碱-O-磷酸伴随着一个特别高的营业额（kcat = 1010 s-1）与碱性磷酸酶。
Derivatives of luciferin, D-luciferin methyl ester, D-luciferyl-L-phenylalanine, D-luciferyl-L-N alpha-arginine, D-luciferin-O-sulphate and D-luciferin-O-phosphate, were synthesized for use as highly sensitive substrates for enzyme assays. The luciferin derivatives were characterized by ultraviolet and fluorescence spectrophotometry, by amino acid analysis and by fast atom bombardement mass spectrometry. Enzymatic cleavage of the compounds by enzymes leading to the release of D-luciferin was demonstrated. Kinetic constants were determined for the following enzyme/substrate pairs: D-luciferin methyl ester/carboxylic esterase, D-luciferyl-L-phenylalanine/carboxypeptidase A, D-luciferyl-L-N alpha-arginine/carboxypeptidase B, D-luciferin-O-sulphate/arylsulphatase, D-luciferin-O-phosphate/alkaline phosphatase. All compounds proved to be acceptable substrates for the respective enzymes, D-luciferin-O-phosphate being accompanied by an especially high turnover number (kcat = 1010 s-1) with alkaline phosphatase.