Transaminase polypeptides
View Patent ↗The present disclosure provides engineered transaminase enzymes having improved properties as compared to a naturally occurring wild-type transaminase enzyme. Also provided are polynucleotides encoding the engineered transaminase enzymes, host cells capable of expressing the engineered transaminase enzymes, and methods of using the engineered transaminase enzymes to synthesize a variety of chiral compounds.
1. An engineered transaminase enzyme comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:2 and has an amino acid residue substitution at X153, wherein the enzyme has transaminase activity.
2. The engineered transaminase enzyme of claim 1 , wherein the amino acid residue substitution X153 of SEQ ID NO:2 is A, C, G, N, M, Q, S, or T.
3. The engineered transaminase enzyme of claim 1 , wherein the amino acid sequence further comprises at least one combination of substitutions selected from the group consisting of:
X57 is F, X86 is H and X153 is Q;
X57 is F, X86 is F and X153 is Q;
X57 is L, X86 is S and X153 is A;
X57 is L, X86 is F, X153 is S, X233 is T, and X417 is T;
X57 is A, X153 is S and X318 is G;
X85 is S, X153 is A and X233 is T;
X85 is A, X153 is S and X417 is S;
X86 is H, X153 is A, X228 is G, and X417 is I;
X86 is H, X153 is A and X417 is C;
X86 is H, X153 is S and X417 is C;
X153 is A, X233 is T and X417 is C;
X153 is C, X233 is L and X318 is R;
X153 is S, X228 is G and X417 is V;
X153 is T, X228 is G and X417 is A;
X153 is A, X317 is L and X318 is G;
X153 is S, X317 is L and X417 is C; and
X153 is S, X318 is R and X417 is E.
4. The engineered transaminase enzyme of claim 1 , wherein enzyme has increased transaminase activity as compared to native SEQ ID NO:2 for conversion of an amino acceptor substrate to a corresponding chiral amino product.
5. The engineered transaminase enzyme of claim 1 , wherein the enzyme has at least 10% residual activity when converting pyruvate to L-alanine in the presence of amino donor isopropylamine after treatment of the polypeptide at 50° C. for 23 h.