Schoevaart R, van Rantwijk F, Sheldon R A
Laboratory of Organic Chemistry and Catalysis, Delft University of Technology, Julianalaan 1362628 BL Delft, The Netherlands.
Biotechnol Bioeng. 2000 Nov 5;70(3):349-52. doi: 10.1002/1097-0290(20001105)70:3<349::aid-bit12>3.0.co;2-0.
A coupled enzymatic assay was developed for quantitative determination of the stereoisomeric products formed in aldol reactions catalyzed by dihydroxyacetone phosphate (DHAP)-dependent aldolases. Three of the four stereoisomers could be determined directly; the fourth one was calculated. This procedure is based on the reversibility of the aldol reaction and requires no derivatization or work-up of the product samples, only removal or inactivation of the biocatalyst. In comparison with other methods the enzymatic assay is highly accurate and fast. Determination of isomer formation with 10 different acceptor substrates applying this procedure gave unprecedented insight in the stereochemistry of fructose-1,6-bisphosphate aldolase from Staphylococcus carnosus and l-rhamnulose-1-phosphate aldolase from E. coli.
开发了一种耦合酶测定法，用于定量测定由磷酸二羟丙酮（DHAP）依赖性醛缩酶催化的醛醇反应中形成的立体异构产物。四种立体异构体中的三种可以直接测定；第四种通过计算得出。该方法基于醛醇反应的可逆性，无需对产物样品进行衍生化或后处理，只需去除生物催化剂或使其失活。与其他方法相比，酶测定法具有高度准确性和快速性。应用该方法对10种不同的受体底物进行异构体形成的测定，为肉葡萄球菌果糖-1,6-二磷酸醛缩酶和大肠杆菌L-鼠李糖-1-磷酸醛缩酶的立体化学提供了前所未有的见解。
