OPTIMIZATION OF ALKYL BETA-D-GALACTOPYRANOSIDE SYNTHESIS FROM LACTOSE USING COMMERCIALLY AVAILABLE BETA-GALACTOSIDASES

被引:60
作者
STEVENSON, DE
STANLEY, RA
FURNEAUX, RH
机构
[1] Industrial Research Ltd, DSIR, Lower Hutt
关键词
BETA-GALACTOSIDASE; LACTOSE; BETA-GALACTOPYRANOSIDE; SYNTHESIS; ORGANIC SOLVENT;
D O I
10.1002/bit.260420514
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Commercially available lactase (beta-D-galactoside galactohydrolase, EC 3.2.1.23) enzymes produced from Kluyveromyces fragilis and Kluyveromyces lactis were accessed as catalysts for use in the production of beta-galactopyranosides of various alcohols using lactose as galactosyl donor. The yield of galactoside was enhanced by using the highest practical concentrations of both lactose and alcohol acceptor. The concentrations and thus yield, were limited by the solubility of the substrates. The increase in galactoside yield with increasing lactose concentration appeared to be specific to the lactose substrate and not due to water activity alterations, because addition of maltose to a fixed concentration of lactose had no effect. During the course of the reaction, the yield of galactoside peaked after around 70% to 80% of the lactose was consumed, due to hydrolysis of the product by the enzyme. A wide variety of compounds with primary or secondary hydroxyl groups could act as acceptors, the essential requirement being at least some water solubility. Addition of organic cosolvents had little effect on galactoside yield except when it increased the water solubility of sparingly soluble alcohols. Some galactosides were synthesized on a gram scale to determine practical product recoveries and improve purification methods for large-scale synthesis. Initial purification by hydrophobic chromatography (for galactosides of hydrophobic alcohols) or strong anion-exchange chromatography (for galactosides of hydrophilic alcohols) separated galactosides, galactobiosides, and higher oligomers from reducing sugars. A facile separation of the galactoside and galactobioside could then be effected by flash chromatography on silica gel. (C) 1993 John Wiley & Sons, Inc.
引用
收藏
页码:657 / 666
页数:10
相关论文
共 29 条
[1]   THE SYNTHESIS OF OLIGOSACCHARIDES BY THE REVERSED HYDROLYSIS REACTION OF BETA-GLUCOSIDASE AT HIGH SUBSTRATE CONCENTRATION AND AT HIGH-TEMPERATURE [J].
AJISAKA, K ;
NISHIDA, H ;
FUJIMOTO, H .
BIOTECHNOLOGY LETTERS, 1987, 9 (04) :243-248
[2]   REGIOSELECTIVE SYNTHESIS OF TRISACCHARIDES BY USE OF A REVERSED HYDROLYSIS ACTIVITY OF ALPHA-D-GALACTOSIDASE AND BETA-D-GALACTOSIDASE [J].
AJISAKA, K ;
FUJIMOTO, H .
CARBOHYDRATE RESEARCH, 1989, 185 (01) :139-146
[3]   ENZYMATIC-SYNTHESIS OF GALACTOPYRANOSYL-L-SERINE DERIVATIVES USING GALACTOSIDASES [J].
CANTACUZENE, D ;
ATTAL, S .
CARBOHYDRATE RESEARCH, 1991, 211 (02) :327-331
[4]   ENZYMATIC-SYNTHESIS OF GLYCOSIDES USING THE BETA-GALACTOSIDASE OF ESCHERICHIA-COLI - REGIOCHEMICAL AND STEREOCHEMICAL STUDIES [J].
CROUT, DHG ;
MACMANUS, DA ;
CRITCHLEY, P .
JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 1, 1990, (07) :1865-1868
[5]   2-BROMOETHYL GLYCOSIDES .3. APPLICATIONS IN THE SYNTHESIS OF SPACER-ARM GLYCOSIDES [J].
DAHMEN, J ;
FREJD, T ;
GRONBERG, G ;
LAVE, T ;
MAGNUSSON, G ;
NOORI, G .
CARBOHYDRATE RESEARCH, 1983, 118 (JUL) :292-301
[6]   DISTORTIONLESS ENHANCEMENT OF NMR SIGNALS BY POLARIZATION TRANSFER [J].
DODDRELL, DM ;
PEGG, DT ;
BENDALL, MR .
JOURNAL OF MAGNETIC RESONANCE, 1982, 48 (02) :323-327
[7]   PRIMARY STRUCTURE OF 10 GALACTOSIDES FORMED BY TRANSGLYCOSYLATION DURING LACTOSE HYDROLYSIS BY BIFIDOBACTERIUM-BIFIDUM [J].
DUMORTIER, V ;
MONTREUIL, J ;
BOUQUELET, S .
CARBOHYDRATE RESEARCH, 1990, 201 (01) :115-123
[8]   LITHIUM-CHLORIDE EFFECT ON PHENYLETHYL-B-D-GALACTOSIDE SYNTHESIS BY ASPERGILLUS-ORYZAE B-D-GALACTOSIDASE IN THE PRESENCE OF HIGH LACTOSE CONCENTRATION [J].
FORTUN, Y ;
COLAS, B .
BIOTECHNOLOGY LETTERS, 1991, 13 (12) :863-866
[9]   DIASTEREOSELECTIVE D-GALACTOPYRANOSYL TRANSFER TO MESO DIOLS CATALYZED BY BETA-GALACTOSIDASES [J].
GAIS, HJ ;
ZEISSLER, A ;
MAIDONIS, P .
TETRAHEDRON LETTERS, 1988, 29 (45) :5743-5744
[10]   BINDING AND REACTIVITY AT THE GLUCOSE SITE OF GALACTOSYL-BETA-GALACTOSIDASE (ESCHERICHIA-COLI) [J].
HUBER, RE ;
GAUNT, MT ;
HURLBURT, KL .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1984, 234 (01) :151-160