ACTIVE-SITE PROTECTION OF PROTEOLYTIC-ENZYMES BY POLY(ETHYLENE GLYCOL) SURFACE MODIFICATION

被引:39
作者
CALICETI, P [1 ]
SCHIAVON, O [1 ]
SARTORE, L [1 ]
MONFARDINI, C [1 ]
VERONESE, FM [1 ]
机构
[1] UNIV PADUA,CNR,CTR STUDIO CHIM FARM & PRODOTTI BIOL ATTIVI,DEPT PHARMACEUT SCI,VIA F MARZOLO 5,I-35121 PADUA,ITALY
关键词
D O I
10.1177/088391159300800103
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
A method to prevent the loss of enzymatic activity of proteolytic enzymes toward high molecular weight substrates that occurs upon derivatization with monomethoxypoly(ethylene glycol) (mPEG) is described. It is based on the heterogenous phase enzyme modification after the enzyme is bound to an active site inhibitor immobilized on an insoluble resin. This procedure protects the active site and the surrounding area from mPEG linkage. Trypsin modified by mPEG in a heterogeneous phase, using benzamidine bound to Sepharose maintained a high degree of its ability to hydrolize large molecular weight substrates, such as bovine serum albumin or casein, compared to the mPEG derivatives obtained without any protection or with free benzamidine in solution.
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页码:41 / 50
页数:10
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