Chaubey Asha, Parshad Rajinder, Gupta Pankaj, Taneja Subhash C, Qazi Ghulam N, Rajan C R, Ponrathnam S
Indian Institute of Integrative Medicine, Canal Road, Jammu Tawi 180001, India.
Bioorg Med Chem. 2009 Jan 1;17(1):29-34. doi: 10.1016/j.bmc.2008.11.023. Epub 2008 Nov 18.
Recent reports on immobilization of lipase from Arthrobacter sp. (ABL, MTCC 5125; IIIM isolate) on insoluble polymers have shown altered properties including stability and enantioselectivity. Present work demonstrates a facile method for the preparation of enantiopure beta-amino alcohols by modulation of ABL enzyme properties via immobilization on insoluble as well as soluble supports using entrapment/covalent binding techniques. Efficacies of immobilized ABL on insoluble supports prepared from tetraethylorthosilicate/aminopropyltriethoxy silane and soluble supports derived from copolymerization of N-vinyl pyrrolidone-allylglycidyl ether (ANP type)/N-vinyl pyrrolidone-glycidyl methacrylate (GNP type) for kinetic resolution of masked beta-amino alcohols have been studied vis-à-vis free ABL enzyme/wet cell biomass. The immobilized lipase on different insoluble/soluble supports has shown 21-110 mg/g protein binding and 30-700 U/g activity for hydrolyzing tributyrin substrate. The findings have shown a significant enhancement in enantioselectivity (ee 99%) vis-à-vis wet cell biomass providing ee 70-90% for resolution of beta-amino alcohols.
最近关于将节杆菌属脂肪酶（ABL，MTCC 5125；IIIM分离株）固定在不溶性聚合物上的报道显示，其性质发生了改变，包括稳定性和对映选择性。目前的工作展示了一种简便的方法，通过使用包埋/共价结合技术将ABL酶固定在不溶性和可溶性载体上，调节其酶性质，从而制备对映体纯的β-氨基醇。研究了通过原硅酸四乙酯/氨丙基三乙氧基硅烷制备的不溶性载体以及由N-乙烯基吡咯烷酮-烯丙基缩水甘油醚（ANP型）/N-乙烯基吡咯烷酮-甲基丙烯酸缩水甘油酯（GNP型）共聚得到的可溶性载体固定化ABL对掩蔽β-氨基醇进行动力学拆分的效果，并与游离ABL酶/湿细胞生物质进行了对比。固定在不同不溶性/可溶性载体上的脂肪酶对三丁酸甘油酯底物的水解显示出21 - 110 mg/g蛋白质结合量和30 - 700 U/g活性。研究结果表明，相对于湿细胞生物质，对映选择性有显著提高（ee为99%），湿细胞生物质对β-氨基醇拆分的ee为70 - 90%。
