Patent ID: 8148128

Claim:
A modified subtilisin that catalyzes a transamidation or a transpeptidation or a transesterification reaction and has a replacement by cysteine of one or more amino acid residues at residue positions selected from the group consisting of residue position 156 in the S1 subsite, residue position 166 in the S1 subsite, residue position 217 in the S1′ subsite, and residue position 62 in the S2 subsite, wherein the amino acid residue positions are numbered by correspondence with the residue positions in the amino acid sequence of Bacillus amyloliquifaciens subtilisin, and wherein the replacement cysteine is modified by replacing the thiol hydrogen in the cysteine with a substituent selected from the group consisting of (R)-2-methoxy-2-phenyl-ethylmethanethiosulfonate, (S)-2-methoxy-2-phenyl-ethylmethanethiosulfonate, (R)-2-hydroxy-2-phenyl-ethylmethanethiosulfonate, (S)-2-hydroxy-2-phenyl-ethylmethanethiosulfonate, N-(3′-methanethiosulfonatopropyl)-2-oxazoldinone, N-(3′-methanethiosulfonatopropyl)-(R)-4-isopropyl-2-oxazolidinone, N-(3′-methanethiosulfonatopropyl)-(S)-4-isopropyl-2-oxazolidinone, N-(3′-methanethiosulfonatopropyl)-(R)-4-phenyl-2-oxazolidinone, N-(3′-methanethiosulfonatopropyl)-(S),-4-phenyl-2-oxazolidinone, N-(3′-methanethiosulfonatopropyl)-(R)-4-benzyl-2-oxazolidinone, N-(3′-methanethiosulfonatopropyl)-(S)-4-benzyl-2-oxazolidinone, N-(2′-methanethiosulfonatoethyl)-(R)-4-phenyl-2-oxazolidinone, N-(2′-methanethiosulfonatoethyl)-(S)-4-phenyl-2-oxazolidinone, N-(2′-methanethiosulfonatopropyl)-(R)-4-benzyl-2-oxazolidinone, N-(2′-methanethiosulfonatopropyl)-(S)-4-benzyl-2-oxazolidinone, N-(3′-methanethiosulfonatopropyl)-(3aR-cis)-3,3a-8,8a-tetrahydro-2H-indeno[1,2-d]-oxazol-2-one, and N-(3′-methanethiosulfonatopropyl)-(3aS-cis)-3,3a-8,8a-tetrahydro-2H-indeno[1,2-d]-oxazol-2-one.