Biosynthesis of beta-lactam antibiotics
OwnerKrupanandan Haranahalli, Shurong Hou +6inventors
InventorsKRUPANANDAN HARANAHALLI, SHURONG HOU, JONATHAN LEWIS MCMURRY, DYLAN STILES, ABRAHAM WALDMAN, EMILY E. WRENBECK, MORTEN MARTINUS CORNELIS HARALD VAN SCHIE, THOMAS VAN DER DOES
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Abstract
Penicillin G acylase (PGA) enzymes, including engineered PGA enzymes, and their use in catalyzing chemical reactions related to biosynthesis of beta-lactam antibiotics.
Claims36
A penicillin G acylase (PGA), wherein the PGA comprises amino acid substitutions relative to the sequence of SEQ ID NO:
- 1 selected from the group consisting of:
- (i) F330A and W333I;
- (ii) M182I and F330A;
- (iii) T90V, A189V, F330A, W333I and D380V;
- (iv) T90V, A189V, F330A, D380V, H498L and V563Q;
- (v) M182L, F330A and T482P;
- (vi) T90V, A189V, F330A, W333I, D380I and H498L;
- (vii) M182T, F330A and L362F;
- (viii) M182L, R185F, F330A and T482P;
- (ix) F330G;
- (x) F330A and W333Y;
- (xi) R185F, N326A, F330A and T482P;
- (xii) F330A and W333P;
- (xiii) F330A and L770T;
- (xiv) T181M and F330A;
- (xv) F330A and D380Q;
- (xvi) M182N and F330A;
- (xvii) R185F, F330A and T482P;
- (xviii) F330A and L770A;
- (xix) T90V, A189V, F330A, W333T, D380V and H498S;
- and
- (xx) T181M, M182F, F330A and L362F.
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12-16. (canceled)
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25-26. (canceled)
28-31. (canceled)
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38-63. (canceled)
65-74. (canceled)
A method of converting 6-aminopenicillanic acid (6-APA) and D-4-hydroxyphenylglucine methyl ester (D-HPGM) to D-amoxicillin, comprising contacting 6-APA and D-HPGM with a penicillin G acylase (PGA), wherein the PGA comprises amino acid substitution relative to the sequence of SEQ ID NO:
- 1 selected from the group consisting of:
- (i) F330A and W333I;
- (ii) M182I and F330A;
- (iii) T90V, A189V, F330A, W333I and D380V;
- (iv) T90V, A189V, F330A, D380V, H498L and V563Q;
- (v) M182L, F330A and T482P;
- (vi) T90V, A189V, F330A, W333I, D380I and H498L;
- (vii) M182T, F330A and L362F;
- (viii) M182L, R185F, F330A and T482P;
- (ix) F330G;
- (x) F330A and W333Y;
- (xi) R185F, N326A, F330A and T482P;
- (xii) F330A and W333P;
- (xiii) F330A and L770T;
- (xiv) T181M and F330A;
- (XV) F330A and D380Q;
- (xvi) M182N and F330A;
- (xvii) R185F, F330A and T482P;
- (xviii) F330A and L770A;
- (xix) T90V, A189V, F330A, W333T, D380V and H498S;
- and
- (xx) T181M, M182F, F330A and L362F.
76-78. (canceled)
A method of converting 7-aminodeacetoxycephalosporanic acid (7-ADCA) and D-phenylglycine methyl ester (D-PGM) to cephalexin, comprising contacting 7-ADCA and D-PGM with a penicillin G acylase (PGA), wherein the PGA comprises amino acid substitution relative to the sequence of SEQ ID NO:
- 1 selected from the group consisting of:
- (i) F330A and W333I;
- (ii) M182I and F330A;
- (iii) T90V, A189V, F330A, W333I and D380V;
- (iv) T90V, A189V, F330A, D380V, H498L and V563Q;
- (v) M182L, F330A and T482P;
- (vi) T90V, A189V, F330A, W333I, D380I and H498L;
- (vii) M182T, F330A and L362F;
- (viii) M182L, R185F, F330A and T482P;
- (ix) F330G;
- (x) F330A and W333Y;
- (xi) R185F, N326A, F330A and T482P;
- (xii) F330A and W333P;
- (xiii) F330A and L770T;
- (xiv) T181M and F330A;
- (XV) F330A and D380Q;
- (xvi) M182N and F330A;
- (xvii) R185F, F330A and T482P;
- (xviii) F330A and L770A;
- (xix) T90V, A189V, F330A, W333T, D380V and H498S;
- and
- (xx) T181M, M182F, F330A and L362F.
80-87. (canceled)
Dependencies are inferred from the claim text.