NEW METHODOLOGY FOR TOSYLATION OF HYDROXYLIC SUPPORTS AS EXEMPLIFIED BY THE IMMOBILIZATION OF MICROCOCCAL ENDONUCLEASE ON AGAROSE

被引:3
作者
ALCANTARA, A
BALLESTEROS, A
SINISTERRA, JV
机构
[1] UNIV COMPLUTENSE MADRID,FAC PHARM,DEPT ORGAN & PHARMACEUT CHEM,E-28040 MADRID,SPAIN
[2] CSIC,INST CATALYSIS,E-28006 MADRID,SPAIN
关键词
agarose; hydrolysis of DNA; immobilization of enzymes; Staphylococcal endonuclease; Tosylation of polysaccharides;
D O I
10.1007/BF02921508
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
Improvement have been made in a simplified procedure we previously reported (J.Mol.Catal. (1986),38,227 for the activation of tosyl chloride of supports possessing primary hydroxyl groups. The method is simple, can be completed in less than 90 min, yields a broad range of activation degrees, and, since it involves no toxic reagents, may be used for preparing immobilized enzymes to be utilized in food manufacturing and processing. The immobilization of Stophylococcal Nuclease has been carried out by this method. The insolubilized derivatives are more active than the native enzyme in the hydrolysis of DNA. The thermal stability of nuclease derivatives is greater than that of the native enzyme. These derivatives remain active at 50°C, and the native enzyme, 39°C. The insolubilized nuclease is more stable against organic solvents such as, dimethylsulfoxide (DMSO) or tetrahydroduran (THF) than the native enzyme. © 1990 Humana Press Inc.
引用
收藏
页码:297 / 310
页数:14
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