Substrate assisted catalysis
AssigneeGenentech, Inc.
InventorsPaul J. Carter, James A. Wells
Life of this patent
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Abstract
Novel enzyme mutants are disclosed which are derived from a precursor enzyme by replacing or modifying at least one catalytic functional group of an amino acid residue in a precursor enzyme. Such mutant enzymes have a catalytic preference for substrates which provide the replaced or modified catalytic group or its equivalent such that the substrate together with the enzyme mutant assists in its own catalysis.
Claims14
A process comprising contacting a subtilisin-related protease variant and a modified substrate wherein said variant is not found in nature and is formed by the replacement in a precursor subtilisin-related protease of a catalytic histidine with an amino acid having a side-chain volume less than the side chain volume of histidine, said histidine being catalytically functional when in contact with a selected region of a substrate, wherein said variant is substantially less active catalytically with said substrate as compared to said catalytic activity of said variant with at least one modified substrate, said modified substrate being formed by replacing an amino acid residue in said selected region with histidine.
A process for cleaving a modified substrate containing histidine within a cleavage recognition sequence comprising contacting said modified substrate with a mutant subtilisin selected from the group consisting of H64A subtilisin, H64A:G166A subtilisin, H64A:E156S:G169A:Y217L subtilisin and H64A:E156S:G166A:G169A:Y217L subtilisin.
Dependencies are inferred from the claim text.