Patent
Provided herein are variants of Buttiauxella sp. phytases that may be used in industrial applications including methods for starch liquefaction, alcohol fermentations and for enhancing phosphate digestion in foods and animal feeds.
1. An isolated phytase variant having phytase activity and an amino acid sequence that varies from the amino acid sequence of the wild type Buttiauxella sp. phytase of SEQ ID NO: 1, wherein the amino acid sequence of the phytase variant comprises a variation at one or more positions corresponding to positions 75, 76, 77, 198, 367 or 374 of SEQ ID NO: 1, and wherein the amino acid sequence of the phytase variant is 90% identical to SEQ ID NO: 1.
2. The phytase variant of claim 1 , wherein the phytase variant further comprises one or more additional variations, wherein the additional variations are substitutions corresponding to substitutions in SEQ ID NO: 1 selected from the group consisting of D92A, F164E, F164S, T171V, T171I, G192A, Q256A, Q256E, and Q256P.
3. The phytase variant of claim 2 , wherein the phytase variant comprises one or more substitutions corresponding to substitutions in SEQ ID NO: 1 selected from the group consisting of S75P, Q76R, G77S, K198R, P367L, and A374P.
4. The phytase variant of claim 3 , wherein the phytase variant further comprises one or more additional variations at positions corresponding to positions of SEQ ID NO: 1 selected from the group consisting of positions 26, 37, 89, 134, 160, 176, 178, 188, 190, 207, 209, 211, 235, 261, 270, 303 and 318.
5. The phytase variant of claim 4 , wherein the one or more additional variations are substitutions corresponding to substitutions in SEQ ID NO: 1 selected from the group consisting of K26E, N37Y, A89T, T134I, T134V, H160R, T176K, A178P, S188N, D190E, K207E, K207T, A209S, D211C, A235V, A261E, N270K, I303F and N318D.
6. The phytase variant of claim 5 , wherein the phytase variant further comprises one or more additional variations at positions corresponding to positions of SEQ ID NO: 1 selected from the group consisting of positions 1, 10, 11, 38, 66, 71, 81, 92, 109, 111, 119, 120, 121, 141, 142, 152, 155, 193, 214, 239, 245, 248, 255, 268, 277, 283, 285, 287, 288, 293, 296, 314, 337, 345, 350, 364, 371, 372, 396, 399, 406 and 413.
7. The phytase variant of claim 6 , wherein the one or more additional variations are substitutions corresponding to substitutions in SEQ ID NO: 1 selected from the group consisting of N1S, V10I, E11I, T38S, Q66E, Q71K, T81A, D92E, Q141R, V142L, T152M, D155E, L193Q, D211C, I214V, A235V, N239K, E245D, S248L, V255A, A261E, R268A, R268T, N270K, A277T, N283D, N285K, T287D, E288A, D293G and P296S.
8. An enzyme composition comprising the phytase variant according to claim 1 , further comprising one or more of a glucoamylase, an alpha-amylase, a protease, a pullulanase, an isoamylase, a cellulase, a hemicellulase, a xylanase, a cyclodextrin glycotransferase, a lipase, a laccase, an oxidase, an esterase, a cutinase, another phytase or any combinations thereof.
9. The enzyme composition of claim 8 , wherein an alpha-amylase is included.