Enantioselectivity modulation through immobilization of Arthrobacter sp lipase:: Kinetic resolution of fluoxetine intermediate

被引:65
作者
Chaubey, Asha [1 ]
Parshad, Rajinder [1 ]
Koul, Surrinder [1 ]
Taneja, Subhash C. [1 ]
Qazi, Ghulam N. [1 ]
机构
[1] Reg Res Lab, CSIR, Div Biotechnol, Jammu 180001, India
关键词
immobilization; enantioselectivity; kinetic resolution; Arthrobacter;
D O I
10.1016/j.molcatb.2006.06.011
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Arthrobacter sp. lipase (ABL, MTCC no. 5125) has been identified for its excellent performance in kinetic resolution of a number of drug intermediates. ABL free enzyme provided product 11 and V (ee < 95%) from racemic fluoxetine intermediate (I and IV) compared to its cell biomass in naturally immobilized state (ee < 98%). To overcome this problem and obtaining high enantioselectivity (R isomer ee 99%), ABL enzyme was modulated by immobilization using various methods vis-A-vis substrate modification (Scheme 2). Immobilized enzyme obtained by hydrophobic binding provided 6710-7720 U/g, covalent binding 200 U/g, and sol-gel entrapment 65-110 U/g activity. Substantial improvement in enantioselectivity was obtained using acylates of ethyl 3-hydroxy-3-phenylpropanoate a fluoxetine drug intermediate (R isomer ee from 93 to 99% and E from 43 to 127-473) at 29-45% conversion in fixed time period of 21 h, indicating thereby some change in conformation of ABL immobilized enzyme. The ABL immobilized by covalent binding and sol-gel entrapment has demonstrated reasonable superiority over the free ABL in enantioselectivity, as well as over all rate of hydrolysis. Immobilized enzymes prepared by covalent and entrapment methods have shown excellent operational stability and used for 10 cycles without loss in activity and the technique can be upscaled for process development. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:39 / 44
页数:6
相关论文
共 39 条
[1]  
ANAND NA, 2005, Patent No. NF0266NF2005
[2]   A novel enzyme support derived from aminated silica gel and polysuccinimide: preparation and application for the immobilization of porcine pancreatic lipase [J].
Bai, ZW ;
Zhou, YK .
REACTIVE & FUNCTIONAL POLYMERS, 2004, 59 (01) :93-98
[3]   Controlling lipase enantioselectivity for organic synthesis [J].
Berglund, P .
BIOMOLECULAR ENGINEERING, 2001, 18 (01) :13-22
[4]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[5]   A MODEL FOR INTERFACIAL ACTIVATION IN LIPASES FROM THE STRUCTURE OF A FUNGAL LIPASE-INHIBITOR COMPLEX [J].
BRZOZOWSKI, AM ;
DEREWENDA, U ;
DEREWENDA, ZS ;
DODSON, GG ;
LAWSON, DM ;
TURKENBURG, JP ;
BJORKLING, F ;
HUGEJENSEN, B ;
PATKAR, SA ;
THIM, L .
NATURE, 1991, 351 (6326) :491-494
[6]  
CHAUBEY A, 2006, IN PRESS APPL MICROB
[7]   QUANTITATIVE-ANALYSES OF BIOCHEMICAL KINETIC RESOLUTIONS OF ENANTIOMERS [J].
CHEN, CS ;
FUJIMOTO, Y ;
GIRDAUKAS, G ;
SIH, CJ .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1982, 104 (25) :7294-7299
[8]   Hydrophobic interaction chromatography of Chromobacterium viscosum lipase on polypropylene glycol immobilised on Sepharose [J].
Diogo, MM ;
Silva, S ;
Cabral, JMS ;
Queiroz, JA .
JOURNAL OF CHROMATOGRAPHY A, 1999, 849 (02) :413-419
[9]   Immobilization of lipases by selective adsorption on hydrophobic supports [J].
Fernandez-Lafuente, R ;
Armisén, P ;
Sabuquillo, P ;
Fernández-Lorente, G ;
Guisán, JM .
CHEMISTRY AND PHYSICS OF LIPIDS, 1998, 93 (1-2) :185-197
[10]   Modulation of lipase properties in macro-aqueous systems by controlled enzyme immobilization:: enantioselective hydrolysis of a chiral ester by immobilized Pseudomonas lipase [J].
Fernández-Lorente, G ;
Terreni, M ;
Mateo, C ;
Bastida, A ;
Fernández-Lafuente, R ;
Dalmases, P ;
Huguet, J ;
Guisán, JM .
ENZYME AND MICROBIAL TECHNOLOGY, 2001, 28 (4-5) :389-396