Buttiauxella sp. phytase variants

Abstract

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.

What is claimed is: 
     
         1 - 15 . (canceled) 
     
     
         16 . A phytase variant having phytase activity and an amino acid sequence that varies from the amino acid sequence of wild type  Buttiauxella  sp. phytase (SEQ ID NO: 1), wherein the amino acid sequence of the phytase variant comprises at least one variation as compared with SEQ ID NO: 1, and wherein the phytase sequence variation comprises
 a) N37Y, S75P, A89T, D92A, T134I, H160R, F164E, T171V, T176K, A178P, S188P, G192A, K198R, K207E, A209S, S248L, Q256Y, A261E, N270K, A374P   b) N37Y, G77S, A89T, D92A, T134I, H160R, F164E, T171V, T176K, A178P, S188P, G192A, K198R, K207E, A209S, S248L, Q256Y, A261E, N270K, A374P   c) N37Y, S75P, Q76R, A89T, D92A, T134I, H160R, F164E, T171I, T176K, A178P, S188P, G192A, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, A374P   d) N37Y, A89T, D92A, T134I, F164E, T171V, T176K, A178P, G192A, K207E, A209S, A235V, S248L, Q256P, A261E, N270K, A374P   e) S75P, Q76R, A89T, D92A, T134I, H160R, F164E, T171I, T176K, A178P, S188P, G192A, K207E, A209S, S248L, Q256Y, A261E, N270K, A374P   f) N37Y, Q76R, A89T, D92A, T134I, H160R, F164E, T171I, T176K, A178P, S188P, G192A, K207E, A209S, S248L, Q256Y, A261E, N270K, A374P   g) N37Y, Q76R, A89T, D92A, T134I, F164S, T171V, T176K, A178P, S188P, G192A, K207E, A209S, A235V, S248L, Q256A, A261E, N270K, A374P   h) S75P, A89T, D92A, T134I, F164E, T171V, T176K, A178P, S188P, G192A, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, A374P   i) S75P, Q76R, A89T, D92A, T134I, H160R, F164E, T171V, T176K, A178P, S188P, G192A, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, P367L, A374P   j) N37Y, A89T, D92A, T134I, F164E, T171I, T176K, A178P, G192A, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, A374P   k) N37Y, Q76R, A89T, D92A, T134I, F164E, T171V, T176K, A178P, G192A, K207E, A209S, S248L, Q256Y, A261E, N270K, A374P   l) N37Y, Q76R, A89T, D92A, T134I, F164E, T171V, T176K, A178P, G192A, K207E, A209S, S248L, Q256A, A261E, N270K, A374P   m) N37Y, S75P, Q76R, A89T, D92A, T134I, F164E, T171V, T176K, A178P, K207E, A209S, A235V, S248L, Q256A, A261E, N270K, A374P   n) N37Y, S75P, A89T, D92A, T134I, H160R, F164E, T171V, T176K, A178P, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, A374P   o) N37Y, A89T, D92A, T134I, H160R, F164S, T171I, T176K, A178P, S188P, G192A, K207E, A209S, A235V, S248L, Q256E, A261E, N270K, A374P   p) A89T, D92A, T134I, H160R, F164E, T171V, T176K, A178P, G192A, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, A374P   q) N37Y, S75P, A89T, D92A, T134I, H160R, F164S, T171V, T176K, A178P, S188P, K207E, A209S, S248L, Q256H, A261E, N270K, A374P   r) N37Y, S75P, A89T, D92A, T134I, F164S, T171V, T176K, A178P, S188P, G192A, K207E, A209S, S248L, Q256A, A261E, N270K, A374P   s) S75P, Q76R, A89T, D92A, T134I, H160R, F164E, T171V, T176K, A178P, G192A, K207E, A209S, S248L, Q256A, A261E, N270K, A374P; or   t) N37Y, Q76R, A89T, D92A, T134I, H160R, F164S, T171V, T176K, A178P, G192A, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, A374P.   
     
     
         17 . The phytase variant of  claim 16 , wherein the phytase sequence variation comprises N37Y, S75P, Q76R, A89T, D92A, T134I, H160R, F164E, T171I, T176K, A178P, S188P, G192A, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, A374P. 
     
     
         18 . A phytase which has at least a minimum percent sequence identity and/or percent homology to the phytase of  claim 16 , wherein the minimum percent identity and/or homology is at least 50%, at least 60%, at least 75%, at least 80%, at least 85%, at least 90%, at least 93%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%. 
     
     
         19 . A phytase variant having phytase activity and an amino acid sequence that varies from the amino acid sequence of the wild type  Buttiauxella  sp. phytase (SEQ ID NO: 1), wherein the amino acid sequence of the phytase variant comprises at least one variation as compared with SEQ ID NO: 1, and wherein the variant has a variation at at least any one of the following positions: 70, 193, 197, 221 and 407. 
     
     
         20 . A phytase of  claim 19 , wherein the variant has at least the following variations: N70Y, H193R, F197E, S221P and A407P. 
     
     
         21 - 26 . (canceled) 
     
     
         27 . A nucleic acid encoding a phytase of  claim 16 . 
     
     
         28 . A vector comprising the nucleic acid of  claim 27 . 
     
     
         29 . A host cell comprising the vector of  claim 28 . 
     
     
         30 . A method of producing a phytase variant of  claim 16  in a host cell, comprising
 a) transforming a host cell with a DNA construct comprising the nucleic acid encoding the phytase of  claim 16 , and 
 b) cultivating the transformed host cell in a suitable culture medium. 
 
     
     
         31 . The method according to  claim 19 , wherein the host cell is a fungal cell, a bacterial cell or a plant cell. 
     
     
         32 . A phytase prepared by the method according to  claim 31  or  claim 31 . 
     
     
         33 . An enzyme composition comprising at least one phytase of  claim 16  including additionally 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. 
     
     
         34 . The enzyme composition of  claim 33 , wherein an alpha-amylase is included. 
     
     
         35 - 57 . (canceled)