Mutant polypeptides and uses thereof
AssigneeInvista North America S.A.R.L.
InventorsNadia Kadi, Adriana L. Botes, Mihai Luchian Azoitei, Yih-En A. Ban, Daniela Grabs-Röthlisberger, Alexander Pisarchik, Alexandre Zanghellini, Eric Althoff
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Abstract
The present disclosure provides novel polypeptides with 3-buten-2-ol dehydratase activity, polypeptides with catalytic activity in the conversion of 3-methyl-3-buten-2-ol to isoprene, and crystal structure data for one of such polypeptides. Methods of making and using the polypeptides and their related crystal structure data are also provided.
Claims22
A polypeptide comprising an amino acid sequence with at least 95% amino acid sequence identity to SEQ ID NO:1, wherein said amino acid sequence comprises one to five mutations at the following X positions of SEQ ID NO:1:
- R1-95X96R97-98X99R100-122X123R124-186X187R188-203X204R205-211X212R213-272X273X274X275R276-323X324R325-327X328R329-359X360R361-365X366R367-381X382R383-398, wherein:
- X382 is mutated to a different amino acid selected from Y and W;
- X328 is mutated to a different amino acid selected from I, M, A, G, and V;
- X187 is mutated to a different amino acid selected from L, M, A, G, and V;
- X96 is mutated to a different amino acid selected from L, I, M, A, G, and V;
- X99 is mutated to a different amino acid selected from L, I, M, A, G, and V;
- X123 is mutated to a different amino acid selected from L, I, M, A, and G;
- X204 is mutated to a different amino acid selected from L, I, M, A, and G;
- X212 is mutated to a different amino acid selected from F, Y, and W;
- X273 is mutated to a different amino acid selected from C, M, S, and T;
- X274 is mutated to a different amino acid selected from F, Y, and W;
- X275 is mutated to a different amino acid selected from L, I, M, A, and G;
- X324 is mutated to a different amino acid selected from L, I, M, G, V, E, D, N, and Q;
- X360 is mutated to a different amino acid selected from F, Y, and W;
- X366 is mutated to a different amino acid selected from L, I, M, A, G, V, C, and T;
- and each R is the same as the corresponding amino acid in SEQ ID NO:1;
- wherein the amino acid sequence comprises at least one mutation selected from F382W, L328V, and I187M, optionally all three, optionally comprising the amino acid sequence of SEQ ID NO:462 or SEQ ID NO:462 without the periplasmic tag;
- wherein the polypeptide is capable of converting 3-buten-2-ol to 1,3-butadiene and/or converting 3-methyl-3-buten-2-ol to isoprene,
- and provided that the mutations chosen from one or more of positions X96, X99, X123, and X187 are not the only mutations in SEQ ID NO:1.
A composition comprising one or more polypeptides according to.
Dependencies are inferred from the claim text.