Immobilized penicillin G acylase as reactor and chiral selector in liquid chromatography

被引:53
作者
Massolini, G
Calleri, E
De Lorenzi, E
Pregnolato, M
Terreni, M
Félix, G
Gandini, C
机构
[1] Univ Pavia, Dept Pharmaceut Chem, I-27100 Pavia, Italy
[2] ENSCPB, F-33402 Talence, France
关键词
immobilized enzyme reactor; chiral stationary phase; LC; enantiomer separation; on-line hydrolysis; penicillin G acylase; enzymes;
D O I
10.1016/S0021-9673(01)00850-0
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
In this paper, the use of penicillin G acylase (PGA) as a biocatalyst and as a chiral selector is described. Penicillin G-acylase is an interesting enzyme used in the manufacture of semisynthetic antibiotics and, in particular, in the production of 6-APA by hydrolysis of penicillin G. Five PGA-based HPLC columns have been prepared by using two different silica supports by employing two immobilization methods, namely "in situ" and "in batch". The effects of the immobilization techniques and of different silica pore size on the catalytic properties of the enzyme as well as the applicability of the PGA-bonded stationary phases as chiral selectors for a number of chiral drugs have been investigated. The HPLC columns based on immobilized PGA combine the hydrolytic activity and the chiral recognition properties of PGA, therefore they have been used for the development of a combined reaction-separation system for chiral and achiral substrates. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:147 / 160
页数:14
相关论文
共 32 条
[1]   INFLUENCE OF THE CHEMICAL AND TEXTURAL PROPERTIES OF THE SUPPORT IN THE IMMOBILIZATION OF PENICILLIN G-ACYLASE FROM KLUYVENA-CITROPHILA ON INORGANIC SUPPORTS [J].
APARICIO, J ;
SINISTERRA, JV .
JOURNAL OF MOLECULAR CATALYSIS, 1993, 80 (02) :269-276
[2]   PEN-G ACYLASE CATALYZED RESOLUTION OF PHENYLACETATE ESTERS OF SECONDARY ALCOHOLS [J].
BALDARO, E ;
DARRIGO, P ;
PEDROCCHIFANTONI, G ;
ROSELL, CM ;
SERVI, S ;
TAGLIANI, A ;
TERRENI, M .
TETRAHEDRON-ASYMMETRY, 1993, 4 (05) :1031-1034
[3]   Immobilization of penicillin G acylase on aminoalkylated polyacrylic supports [J].
Bianchi, D ;
Golini, P ;
Bortolo, R ;
Cesti, P .
ENZYME AND MICROBIAL TECHNOLOGY, 1996, 18 (08) :592-596
[4]   Biocatalysis for Pharmaceuticals-Status and Prospects for a Key Technology [J].
Buckland, Barry C. ;
Robinson, David K. ;
Chartrain, Michel .
METABOLIC ENGINEERING, 2000, 2 (01) :42-48
[5]   Cross-linked enzyme aggregates: A simple and effective method for the immobilization of penicillin acylase [J].
Cao, LQ ;
van Rantwijk, F ;
Sheldon, RA .
ORGANIC LETTERS, 2000, 2 (10) :1361-1364
[6]   ENZYME-BASED HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY SUPPORTS AS PROBES OF ENZYME-ACTIVITY AND INHIBITION - THE IMMOBILIZATION OF TRYPSIN AND ALPHA-CHYMOTRYPSIN ON AN IMMOBILIZED ARTIFICIAL MEMBRANE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY SUPPORT [J].
CHUI, WK ;
WAINER, IW .
ANALYTICAL BIOCHEMISTRY, 1992, 201 (02) :237-245
[7]   Riboflavin binding proteins as chiral selectors in HPLC and CE [J].
De Lorenzi, E ;
Massolini, G .
PHARMACEUTICAL SCIENCE & TECHNOLOGY TODAY, 1999, 2 (09) :352-364
[8]   PENICILLIN ACYLASE HAS A SINGLE-AMINO-ACID CATALYTIC CENTER [J].
DUGGLEBY, HJ ;
TOLLEY, SP ;
HILL, CP ;
DODSON, EJ ;
DODSON, G ;
MOODY, PCE .
NATURE, 1995, 373 (6511) :264-268
[9]   Stereochemical resolution of racemates, in HPLC, using a chiral stationary phase based upon immobilized α-chymotrypsin.: II.: Enzyme chiral separations [J].
Félix, G ;
Descorps, V .
CHROMATOGRAPHIA, 1999, 49 (11-12) :606-614
[10]   Stereochemical resolution of racemates, in HPLC, using a chiral stationary phase based upon immobilized α-chymotrypsin.: I.: Structural chiral separations [J].
Félix, G ;
Descorps, V .
CHROMATOGRAPHIA, 1999, 49 (11-12) :595-605