# US 8,541,036 B2 · Synergetic effect of the phytase combination on the hydrolysis of phytic acid > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,541,036 B2 - Document ID: US8541036B2 - Status: Granted - Kind: B2 - Application number: 11/988,403 - Filed: 2006-07-07 - Published: 2010-11-04 - Granted: 2013-09-24 - Earliest claimed: 2005-07-08 - Assignees: Adisseo France S.A.S. - Applicants: Helene Boze; Guy Moulin - Inventors: Helene Boze; Guy Moulin - Web page: https://patentgrep.com/patent/US8541036B2 ## CPC - C12N 9/16: acting on ester bonds (3.1) - A61P 3/00: Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00) - A61P 7/00: Drugs for disorders of the blood or the extracellular fluid - C12P 3/00: Preparation of elements or inorganic compounds except carbon dioxide {(recovery of carbon dioxides as by-products C12F3/02)} - C12P 7/18: polyhydric - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)} ## Abstract The invention concerns compositions and methods combining at least two phytases for hydrolyzing phytic acid (myo-inositol hexakis phosphate) into inorganic monophosphates, into myo-inositols with lower degree of phosphorylation and into free myo-inositol. Said compositions and methods are of particular interest for animal feeding. ## Claims ### Claim 1 A composition combining at least two 3-phytases for hydrolysis of phytic acid (myo-inositol 1,2,3,4,5,6-hexakisphosphate), comprising: a) a first phytase exhibiting at least 95% identity with the phytase of SEQ ID NO: 1 or exhibiting at least 95% identity with the phytase of SEQ ID NO: 2; b) a second phytase exhibiting at least 95% identity with the phytase of SEQ ID NO: 3. wherein the first and second phytases have a synergistic hydrolysis effect on phytic acid in the absence of a phytase having a different stereospecificity. ### Claim 2 (depends on 1) The composition according to claim 1, wherein the first phytase is a 3-phytase which catalyses the hydrolysis of the six phosphate bonds of phytic acid. ### Claim 3 (depends on 1) The composition according to claim 1, wherein the first phytase has an optimum temperature of between 55° C. and 80° C. and an optimum pH of between pH 3.5 and pH 5. ### Claim 4 (depends on 1) The composition according to claim 1, wherein the first phytase is a phytase of the yeast Schwanniomyces castellii or a phytase of the yeast Debaryomyces castellii. ### Claim 5 (depends on 1) The composition according to claim 1, wherein the first phytase is the phytase of SEQ ID NO: 1 or the phytase of SEQ ID NO: 2, and the second phytase is the phytase of SEQ ID NO: 3. ### Claim 6 (depends on 1) The composition according to claim 1, wherein the second phytase is a 3-phytase which catalyses the hydrolysis of at least five phosphate bonds of phytic acid. ### Claim 7 (depends on 1) The composition according to claim 1, wherein the second phytase has an optimum temperature of between 50° C. and 60° C. and an optimum pH of between pH 2 and pH 6. ### Claim 8 (depends on 1) The composition according to claim 1, wherein the second phytase is an Aspergillus niger phytase. ### Claim 9 (depends on 1) The composition according to claim 1, wherein the first phytase exhibits at least 99% identity with the phytase of SEQ ID NO: 1 or exhibits at least 99% identity with the phytase of SEQ ID NO: 2, and the second phytase exhibits at least 99% identity with the phytase of SEQ ID NO: 3. ### Claim 10 (depends on 1) The composition according to claim 1, wherein said composition is in the form of a nutritional additive for animals or of an animal feed. ### Claim 11 (depends on 1) A method for supplementing the nutrition of an animal comprising feeding said animal the composition of claim 1 in a nutritional additive for animals or in an animal feed. ### Claim 12 (depends on 1) A method for increasing the availability of phytic phosphorus and improving the digestibility of animal feeds, comprising feeding animals the composition of claim 1. ### Claim 13 A method of hydrolysis of phytic acid (myo-inositol 1,2,3,4,5,6-hexakisphosphate) to inorganic monophosphates, to myo-inositols with a lower degree of phosphorylation and to free myo-inositol, comprising: hydrolyzing phytic acid with a first phytase and a second phytase, the first phytase exhibiting at least 95% identity with the 3-phytase of SEQ ID NO: 1 or exhibiting at least 95% identity with the 3-phytase of SEQ ID NO: 2, and the second phytase exhibiting at least 95% identity with the 3-phytase of SEQ ID NO: 3, wherein the first and second phytases have a synergistic hydrolysis effect on phytic acid in the absence of a phytase having a different stereospecificity. ### Claim 14 (depends on 13) The method according to claim 13, wherein the first phytase is a 3-phytase which catalyses the hydrolysis of the six phosphate bonds of phytic acid. ### Claim 15 (depends on 13) The method according to claim 13, wherein the first phytase has an optimum temperature of between 55° C. and 80° C. and an optimum pH of between pH 3.5 and pH 5. ### Claim 16 (depends on 13) The method according to claim 13, wherein the first phytase is a phytase of the yeast Schwanniomyces castellii or a phytase of the yeast Debaryomyces castellii. ### Claim 17 (depends on 13) The method according to claim 13, wherein the first phytase is the phytase of SEQ ID NO: 1 or the phytase of SEQ ID NO: 1 and the second phytase is the phytase of SEQ ID NO: 3. ### Claim 18 (depends on 13) The method according to claim 13, wherein the second phytase is a 3-phytase which catalyses the hydrolysis of at least five phosphate bonds of phytic acids. ### Claim 19 (depends on 13) The method according to claim 13, wherein the second phytase has an optimum temperature of between 50° C. and 60° C. and an optimum pH of between pH 2 and pH 6. ### Claim 20 (depends on 13) The method according to claim 13, wherein the second phytase is an Aspergillus niger phytase. ### Claim 21 (depends on 13) The method according to claim 13, wherein the first phytase exhibits at least 99% identity with the phytase of SEQ ID NO: 1 or exhibits at least 99% identity with the phytase of SEQ ID NO: 2, and the second phytase is the exhibits at least 99% identity with the phytase of SEQ ID NO: 3. ### Claim 22 A kit or assembly for feeding animals, comprising: the Schwanniomyces castellii 3-phytase of SEQ ID NO: 1 or the Debaryomyces castellii 3-phytase of SEQ ID NO: 2; and the Aspergillus niger 3-phytase of SEQ ID NO: 3, wherein the Schwanniomyces castellii 3-phytase of SEQ ID NO: 1 or the Debaryomyces castellii 3-phytase of SEQ ID NO: 2, in combination with the Aspergillus niger 3-phytase of SEQ ID NO: 3, have a synergistic hydrolysis effect on phytic acid in the absence of a phytase having a different stereospecificity. ### Claim 23 (depends on 13) The method according to claim 13, wherein phytic acid is completely hydrolyzed. ### Claim 24 (depends on 1) The composition according to claim 1, wherein the synergistic hydrolysis effect on phytic acid is obtained over a pH range of 3-6.5. ### Claim 25 (depends on 1) The composition according to claim 1, wherein the first phytase is the phytase exhibiting at least 95% identity with the phytase of SEQ ID NO: 2. ### Claim 26 (depends on 13) The method according to claim 13, wherein the first phytase is the phytase exhibiting at least 95% identity with the phytase of SEQ ID NO: 2. ### Claim 27 (depends on 22) The kit or assembly according to claim 22, wherein the kit or assembly comprises the Debaryomyces castellii 3-phytase of SEQ ID NO: 2, and wherein the Debaryomyces castellii 3-phytase of SEQ ID NO: 2, in combination with the Aspergillus niger 3-phytase of SEQ ID NO: 3, have a synergistic hydrolysis effect on phytic acid in the absence of a phytase having a different stereospecificity.