DNASE ACTIVITY OF MICROCOCCAL ENDONUCLEASE COVALENTLY IMMOBILIZED ON NYLON AND POLYSTYRENE

被引:9
作者
MICHALON, P
ROCHE, J
COUTURIER, R
FAVREBONVIN, G
MARION, C
机构
[1] UNIV PARIS 06,BIOL STRUCT LAB,BAT C,9 QUAI ST BERNARD,F-75252 PARIS 05,FRANCE
[2] UNIV LYON,CTR GENET MOLEC & CELLULAIRE,VILLEURBANNE,FRANCE
[3] UNIV LYON,BIOCHIM APPL LAB,VILLEURBANNE,FRANCE
[4] UNIV LYON,BIOCHIM ANALYT LAB,VILLEURBANNE,FRANCE
关键词
ENZYME IMMOBILIZATION; ENDONUCLEASES; SYNTHETIC SUPPORTS; NYLON; POLYSTYRENE;
D O I
10.1016/0141-0229(93)90140-W
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
Water-insoluble nucleases were prepared by immobilizing the endonuclease from S. aureus onto the surface of nylon-66 and polystyrene spheres. The activation phase of the synthetic supports was optimized to define optimal conditions of pH, temperature, and Ca2+ concentration for using immobilized enzymes. The activity, evaluated by hydrolysis of high-molecular-weight and supercoiled DNA, indicates that both derivatives are highly stable for storage and further use. Immobilization of the enzyme is much more effective when the covalent binding is performed on polystyrene. By using different activation methods with these matrices, a set of immobilized nucleases with various levels of enzymatic activity can be prepared. The possibility of working in a wide range of enzymatic activity and at low temperature and Ca2+ Concentrations in different buffers makes these immobilized nucleases very useful for investigating accessible DNA regions in chromatin structure.
引用
收藏
页码:215 / 221
页数:7
相关论文
共 19 条
[1]  
Anfinsen C, 1971, ENZYMES, V4, P177
[2]  
COUTURIER R, 1988, LIFE SCI ADV, V7, P107
[3]  
CUATRECA.P, 1967, J BIOL CHEM, V242, P1541
[4]   HYDROLYSIS OF NUCLEIC-ACIDS BY SEPHAROSE-MICROCOCCAL ENDONUCLEASE [J].
GUISAN, JM ;
BALLESTEROS, A .
ENZYME AND MICROBIAL TECHNOLOGY, 1981, 3 (04) :313-320
[5]  
GUISAN JM, 1979, J SOL-PHASE BIOCHEM, V4, P245
[6]   USE OF AN IMMOBILIZED ENZYME AND SPECIFIC ANTIBODIES TO ANALYZE THE ACCESSIBILITY AND ROLE OF HISTONE TAILS IN CHROMATIN STRUCTURE [J].
HACQUES, MF ;
MULLER, S ;
DEMURCIA, G ;
VANREGENMORTEL, MHV ;
MARION, C .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1990, 168 (02) :637-643
[7]   ACCESSIBILITY AND STRUCTURAL ROLE OF HISTONE DOMAINS IN CHROMATIN - BIOPHYSICAL AND IMMUNOCHEMICAL STUDIES OF PROGRESSIVE DIGESTION WITH IMMOBILIZED PROTEASES [J].
HACQUES, MF ;
MULLER, S ;
DEMURCIA, G ;
VANREGENMORTEL, MHV ;
MARION, C .
JOURNAL OF BIOMOLECULAR STRUCTURE & DYNAMICS, 1990, 8 (03) :619-641
[8]   PROTEASE IMMOBILIZATION ONTO POLYACROLEIN MICROSPHERES [J].
HAYASHI, T ;
IKADA, Y .
BIOTECHNOLOGY AND BIOENGINEERING, 1990, 35 (05) :518-524
[9]  
Hornby W E, 1976, Methods Enzymol, V44, P118
[10]  
INAKI Y, 1987, FUNCTIONAL MONOMERS, P461