Variants of thermovibrio ammonificans carbonic anhydrase and CO
The present description relates to recombinant or engineered carbonic anhydrase polypeptides, variants, and functional derivatives thereof, having improved properties that make them advantageous for use in CO 2 capture operations (e.g., CO 2 capture solvents, alkaline pH, and/or elevated temperatures), as well as polynucleotides and vectors encoding same. The present description also relates to methods, processes and systems for CO 2 capture which make use of the recombinant or engineered carbonic anhydrase polypeptides, variants, and functional derivatives thereof.
1. A recombinant carbonic anhydrase polypeptide having carbonic anhydrase activity comprising an amino acid sequence having at least 90% identity to SEQ ID NO: 7, and one or more differences as compared to SEQ ID NO: 7 at residue positions selected from 2, 5, 7, 8, 9, 12, 13, 16, 17, 18, 19, 21, 22, 52, 73, 77, 116, 125, 126, 131, 138, 156, 190, 193, and 206.
2. The recombinant polypeptide of claim 1 , wherein said amino acid difference is at a position corresponding to position 2, 5, 7, 8, 9, 12, 13, 16, 17, 18, 19, 21, 22, or any combination thereof, of SEQ ID NO: 7.
3. The recombinant carbonic anhydrase polypeptide of claim 1 , comprising one or more amino acid differences as compared to SEQ ID NO: 7 selected from: 2Q or 2V; 5C or 5R; 7C, 7F, or 7H; 8C or 8R; 9C, 9P or 9N; 12D, 12R or 12V; G13Q; 16C, 16G, or 16R; 17C; 18V; 19S; 21L, 21D, 21N, 21Q, 21V, or 21G; 22K, 22L, 22P, or 22N; 52I; 73C; 77D; 116N; 125A; 126L; 131I; 138N or 138R; 156E; 190C or 190M; 193S; and 206T.
4. The recombinant carbonic anhydrase polypeptide of claim 1 comprising two or more amino acid differences as compared to SEQ ID NO: 7 which are: 12D and 21Q; 12D and 27T; 12D and 142I; 16C and 167S; 16R and 206T; 16G and 50M; 19S and 138R; 19S and 158G; 150M and 193S; 190M and 193S; 141H and 151C; 8C and 141H; 8C and 151C; 8C, 141H, and 151C; 9P and 21D; 9P and 21N; 9P and 21Q; 9P and 21V; 9P and 21G; 9P and 22P; 8R, 9P, and 22P; 8R, 9P, 22P, and 156E.
5. The recombinant carbonic anhydrase polypeptide of claim 1 , further comprising 227Y and 228G.
6. The recombinant carbonic anhydrase polypeptide of claim 1 , having at least 90% identity to SEQ ID NO:7.
7. The recombinant carbonic anhydrase polypeptide of claim 1 , having at least 95% identity to SEQ ID NO:7.
8. The recombinant carbonic anhydrase polypeptide of claim 1 , comprising one or more amino acid differences as compared to SEQ ID NO: 7 selected from:
(a) 8C or 8R;
(b) 9C, 9P or 9N;
(c) 22K, 22L, 22P, or 22N; or
(d) any combination of (a) to (c),
wherein said recombinant carbonic anhydrase polypeptide exhibits improved stability in a CO2 capture solvent comprising carbonate ions as compared to the polypeptide of SEQ ID NO:7.
9. A recombinant carbonic anhydrase polypeptide having carbonic anhydrase activity comprising:
(a) an amino acid sequence having at least 90% identity to SEQ ID NO:7; and
(b) one or more differences as compared to SEQ ID NO: 7 at residue positions selected from 2, 5, 7, 8, 9, 12, 13, 16, 17, 18, 19, 21, 22, 52, 73, 77, 116, 125, 126, 131, 138, 156, 190, 193, and 206,
wherein said recombinant carbonic anhydrase polypeptide has improved stability relative to the carbonic anhydrase of SEQ ID NO: 7, following 1 hour of exposure in 1.45 M K 2 CO 3 pH 10 at 70° C., 85° C. or 90° C.
10. The recombinant polypeptide of claim 9 , wherein said amino acid difference is at a position corresponding to position 2, 5, 7, 8, 9, 12, 13, 16, 17, 18, 19, 21, 22, or any combination thereof, of SEQ ID NO: 7.
11. The recombinant carbonic anhydrase polypeptide of claim 9 , comprising one or more amino acid differences as compared to SEQ ID NO: 7 selected from: 2Q or 2V; 5C or 5R; 7C, 7F, or 7H; 8C or 8R; 9C, 9P or 9N; 12D, 12R or 12V; G13Q; 16C, 16G, or 16R; 17C; 18V; 19S; 21L, 21D, 21N, 21Q, 21V, or 21G; 22K, 22L, 22P, or 22N; 52I; 73C; 77D; 116N; 125A; 126L; 131I; 138N or 138R; 156E; 190C or 190M; 193S; and 206T.
12. The recombinant carbonic anhydrase polypeptide of claim 9 comprising two or more amino acid differences as compared to SEQ ID NO: 7 which are: 12D and 21Q; 12D and 27T; 12D and 142I; 16C and 167S; 16R and 206T; 16G and 50M; 19S and 138R; 19S and 158G; 150M and 193S; 190M and 193S; 141H and 151C; 8C and 141H; 8C and 151C; 8C, 141H, and 151C; 9P and 21D; 9P and P21N; 9P and 21Q; 9P and 21V; 9P and 21G; 9P and 22P; 8R, 9P, and 22P; 8R, 9P, 22P, and 156E.
13. The recombinant carbonic anhydrase polypeptide of claim 9 , further comprising 227Y and 228G.
14. The recombinant carbonic anhydrase polypeptide of claim 9 , having at least 85% identity to SEQ ID NO: 7.
15. The recombinant carbonic anhydrase polypeptide of claim 9 , having at least 90% identity to SEQ ID NO: 7.
16. The recombinant carbonic anhydrase polypeptide of claim 9 , having at least 95% identity to SEQ ID NO: 7.
17. The recombinant carbonic anhydrase polypeptide of claim 9 , comprising one or more amino acid differences as compared to SEQ ID NO: 7 selected from:
(a) 8C or 8R;
(b) 9C, 9P or 9N;
(c) 22K, 22L, 22P, or 22N; or
(d) any combination of (a) to (c).
18. A recombinant variant of a Thermovibrio ammonificans carbonic anhydrase polypeptide, said Thermovibrio ammonificans carbonic anhydrase polypeptide comprising an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 7, the variant having carbonic anhydrase activity and comprising one or more amino add differences as compared to SEQ ID NO: 7 at residue positions selected from 2, 5, 7, 8, 9, 12, 13, 16, 17, 18, 19, 21, 22, 52, 73, 77, 116, 125, 126, 131, 138, 156, 190, 193, and 206, wherein said variant has improved stability relative to the carbonic anhydrase of SEQ ID NO: 7, following 1 hour of exposure in 1.45 M K 2 CO 3 pH 10 at 70° C., 85° C. or 90° C.
19. The recombinant carbonic anhydrase polypeptide of claim 18 , comprising one or more amino acid differences as compared to SEQ ID NO: 7 selected from: 2Q or 2V; 5C or 5R; 7C, 7F, or 7H; 8C or 8R; 9C, 9P or 9N; 12D, 12R or 12V; G13Q; 16C, 16G, or 16R; 17C; 18V; 19S; 21L, 21D, 21N, 21Q, 21V, or 21G; 27T; 50M; 52I; 73C; 77D; 116N; 125A; 126L; 131I; 138N or 138R; 141H; 142I; 150M; 151C; 156E; 158G; 167S; 173C; 181Y; 190C or 190M; 193S; and 206T.
20. The recombinant carbonic anhydrase polypeptide of claim 18 , comprising two or more amino acid differences as compared to SEQ ID NO: 7 which are: 12D and 21Q; 12D and 27T; 12D and 142I; 16C and 167S; 16R and 206T; 16G and 50M; 19S and 138R; 19S and 158G; 150M and 193S; 190M and 193S; 141H and 151C; 8C and 141H; 8C and 151C; 8C, 141H, and 151C; 9P and 21D; 9P and P21N; 9P and 21Q; 9P and 21V; 9P and 21G; 9P and 22P; 8R, 9P, and 22P; 8R, 9P, 22P, and 156E.