Amylase polypeptides
View Patent ↗This invention relates to polypeptides, more specifically amylase polypeptides and nucleic acids encoding these, and their uses e.g. as non-maltogenic exoamylases in producing food or feed products.
1. A recombinantly produced polypeptide having amylase activity comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 1, and wherein the polypeptide comprises one or more amino acid substitutions at the following positions: 235, 240, 311, 350, 364, or 409 with reference to the position numbering of the sequence shown in SEQ ID NO: 1.
2. The polypeptide according to claim 1 , wherein the polypeptide further comprises:
one or more of the following amino acids 33Y, 34N, 70D, 121F, 134R, 141P, 146G, 157L, 161A, 178F, 179T, 223E/S/K/A, 229P, 307K, 309P and 334P;
one or more amino acid substitutions at positions 88 or 205;
one or more of the following amino acid substitutions: 42K/A/V/N/I/H/F, 34Q, 100Q/K/N/R, 272D, 392 K/D/E/Y/N/Q/R/T/G or 399C/H;
one or more amino acid substitutions at positions 44, 96, 204, 354 or 377; or
a substitution at position 392S.
3. The polypeptide according to claim 2 , wherein the polypeptide comprises one or more amino acid substitutions at the following positions: 235, 88, 205, 240, 248, 266, 311, 377 or 409 and one or more of the following amino acid substitutions: 42K/A/V/N/I/H/F, 34Q, 100Q/K/N/R, 272D or 392K/D/E/Y/N/Q/R/S/T/G.
4. The polypeptide according to claim 2 , wherein the polypeptide comprises one or more amino acid substitutions at the following positions: 235, 88, 205, 240, 311 or 409 and one or more of the following amino acid substitutions: 42K/N/I/H/F, 272D, or 392 K/D/E/Y/N/Q/R/S/T/G.
5. The polypeptide according to claim 2 , wherein the polypeptide comprises amino acid substitutions at least in four, five or in all of the following positions: 88, 205, 235, 240, 311 or 409 and has at least one or two of the following amino acid substitutions: 42K/N/I/H/F, 272D or 392 K/D/E/Y/N/Q/R/S/T/G.
6. The polypeptide according to claim 1 having 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 1.
7. The polypeptide according to claim 1 , wherein the polypeptide further comprises an amino acid substitution in position 88.
8. The polypeptide according to claim 7 , wherein the polypeptide has the amino acid 88L.
9. The polypeptide according to claim 1 , wherein the polypeptide comprises an amino acid substitution in position 235.
10. The polypeptide according to claim 9 , wherein the polypeptide has the amino acid 235R.
11. The polypeptide according to claim 1 , wherein the polypeptide further comprises one or more of the following amino acids 121F, 134R, 141P, 229P, or 307K.
12. The polypeptide according to claim 1 , comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33 and SEQ ID NO: 34.
13. The polypeptide according to claim 1 having a linker fused at the C-terminus.
14. The polypeptide according to claim 1 having exoamylase activity.
15. The polypeptide according to claim 1 having non-maltogenic exoamylase activity.
16. A food or feed additive comprising the polypeptide of claim 1 .
17. The polypeptide according to claim 2 , comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33 and SEQ ID NO: 34.
18. A food, feed product, dough product or bakery product, comprising a polypeptide of claim 1 .
19. A method of making a saccharide syrup, comprising adding a polypeptide according to claim 1 to a granular starch liquefact to form the saccharide syrup.