Clermont Université, Université Blaise Pascal, Laboratoire SEESIB, BP 10448, F-63000 Clermont-Ferrand, France.
J Biotechnol. 2010 Feb 15;145(4):359-66. doi: 10.1016/j.jbiotec.2009.12.022. Epub 2010 Jan 14.
5-O-Coumarinyl-d-xylulose was studied as a fluorogenic substrate for the stereospecific assay of transketolase enzyme. Enzymatic C2-C3 cleavage released an alpha-hydroxyl, beta-coumarinyl substituted aldehyde. Although the subsequent beta-elimination step was rate limiting under chemical or enzymatic catalysis, we detected a TK activity as low as 0.7mIU. To improve the fluorescence signal release, kinetic and product distribution analyses of this reaction were performed by LC/UV/MS coupling.
5-O-香豆酰基-D-木酮糖被研究为酮醇酶立体特异性测定的荧光底物。酶促 C2-C3 裂解释放出 α-羟基、β-香豆酰取代的醛。尽管在化学或酶催化下，随后的β消除步骤是限速步骤，但我们检测到的 TK 活性低至 0.7mIU。为了提高荧光信号释放，通过 LC/UV/MS 偶联对该反应进行了动力学和产物分布分析。
