Sotolongo V, Johnson D V, Wahnon D, Wainer I W
Department of Oncology, McGill University, Montreal, Quebec, Canada.
Chirality. 1999;11(1):39-45. doi: 10.1002/(SICI)1520-636X(1999)11:1<39::AID-CHIR7>3.0.CO;2-L.
Horse liver alcohol dehydrogenase (HLADH) has been non-covalently immobilized on an immobilized artificial membrane (IAM) high-performance liquid chromatography (HPLC) stationary phase. The resulting IAM-HLADH retained the reductive activity of native HLADH as well as the enzyme's enantioselectivity and enantiospecificity. HLADH was also immobilized in an IAM HPLC stationary phase prepacked in a 13 x 4.1 mm ID column to create an immobilized enzyme reactor (HLADH-IMER). The reactor was connected through a switching valve to a column containing a chiral stationary phase (CSP) based upon p-methylphenylcarbamate derivatized cellulose (Chiralcel OJR-CSP). The results from the combined HLADH-IMER/CSP and chromatographic system demonstrate that the enzyme retained its activity and stereoselectivity after immobilization in the column and that the substrate and products from the enzymatic reduction could be transferred to a second column for analytical or preparative separation. The combined HLADH-IMER/CSP system is a prototype for the preparative on-line use of cofactor-dependent enzymes in large-scale chiral syntheses.
马肝醇脱氢酶（HLADH）已通过非共价方式固定在固定化人工膜（IAM）高效液相色谱（HPLC）固定相上。所得的IAM - HLADH保留了天然HLADH的还原活性以及该酶的对映体选择性和对映体特异性。HLADH还被固定在一个内径为13×4.1 mm的预填充IAM HPLC固定相中，以创建一个固定化酶反应器（HLADH - IMER）。该反应器通过一个切换阀连接到一根装有基于对甲基苯基氨基甲酸酯衍生化纤维素的手性固定相（CSP）（Chiralcel OJR - CSP）的柱子上。HLADH - IMER/CSP组合色谱系统的结果表明，该酶在固定于柱中后仍保留其活性和立体选择性，并且酶促还原反应的底物和产物可以转移到第二根柱子上进行分析或制备分离。HLADH - IMER/CSP组合系统是大规模手性合成中依赖辅因子的酶在线制备应用的一个原型。
