1. Protease Activity on an Immobilized Substrate Modified by Polymers: Subtilisin BPN‘
- Author
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and Channing R. Robertson, Giuseppe Trigiante, Philip Frederick Brode, Alan R. Esker, Deborah S. Rauch, Alice P. Gast, Hyuk Yu, and Donn N. Rubingh
- Subjects
Serine protease ,chemistry.chemical_classification ,Protease ,Ethylene oxide ,biology ,Chemistry ,medicine.medical_treatment ,Surfaces and Interfaces ,Polymer ,Condensed Matter Physics ,Hydrolysis ,chemistry.chemical_compound ,Adsorption ,Covalent bond ,Polymer chemistry ,Electrochemistry ,medicine ,biology.protein ,Copolymer ,Organic chemistry ,General Materials Science ,Spectroscopy - Abstract
We describe the adsorption and catalytic behavior of the serine protease subtilisin BPN‘ on controlled pore glass (CPG) beads with a short (aminopropyl) or a long (aminoalkyl CH2 > 12) chain covalent link separating the reporter peptide succinyl-alanine-alanine-proline-phenylalanine-p-nitroanilide (sAAPFpNA) from the surface. The propyl-linked sAAPFpNA modified glass surface (aminopropyl CPG:sAAPFpNA) showed a 2-fold increase in protease adsorption over an aminopropyl−glass surface. In contrast, the sAAPFpNA surface with the long chain connector showed a 2-fold drop in adsorption relative to an aminoalkyl surface. BPN‘-catalyzed hydrolysis rates showed an inverse relationship to adsorption. Water-soluble polymers [poly(vinylpyrrolidone) (PVP), poly(ethylene oxide) (PEO), poly(4-vinylpyridine-N-oxide) (PVPO) and a copolymer of 1-vinyl-2-pyrrolidone and 1-vinylimidazole (PVPVI)] neutralize the 2-fold increase in BPN‘ adsorption and provide more than a 3-fold increase in the initial rate of hydrolysis for BP...
- Published
- 2000
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