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Substrate and the enzyme fixation manner and enzyme desorption manner null for enzyme fixation
Substrate and the enzyme fixation manner and enzyme desorption manner null for enzyme fixation
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机译:酶固定的底物和酶固定方式及酶解吸方式无效
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摘要
PURPOSE:To provide the subject carrier, enzyme immobilization process and desorption process so designed that an enzyme is bonded to the primary amine of a specific carrier through gultaraldehyde(GLA) as the crosslinking agent to retain the activity of the enzyme at high level for a long period and facilitate the desorption of said enzyme leading to improved economy. CONSTITUTION:First, (A) the objective carrier consisting, as the matrix, of a basic anion exchange resin of huge network structure with an average granular size of 0.2-1mm, pore size of 100-2,000Angstrom and pore volume of 0.5-1.5ml/g can be obtained by adding, as the functional group, e.g., ethanolamine to a matrix prepared by polymerization of a methacrylic glycidyl ester or the like. Secondary, the carrier A is buffered with (B) a pH buffer solution for stabilizing the enzyme to be used, and is brought into contact with the solution B containing 0.5-10wt.% of GLA at 4-40 deg.C for 0.5-2hrs. followed by washing an excess of the GLA off with the solution B. Thirdly, the resultant carrier A is brought into contact with (C) an enzyme (e.g., protease) solution for =1hr. to immobilize said enzyme so as to be 0.5-50mg/ml carrier in the enzyme concentration. Finally, when needed, the resultant immobilized enzyme is brought into contact with 2-5wt.% aq. NaOH solution or the like at 40-70 deg.C for 0.5hrs. to effect desorption of the enzyme C from the carrier A.
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