Phytase variants and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-12037613-B2
Application number	US-202117402099-A
Country	US
Kind code	B2
Filing date	Aug 13, 2021
Priority date	Aug 13, 2020
Publication date	Jul 16, 2024
Grant date	Jul 16, 2024

Abstract

The present invention relates to phytase variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

First claim

The invention claimed is:

1. A phytase variant which has phytase activity and an amino acid sequence having at least 70% identity to SEQ ID NO: 2, which comprises alterations N31C/G52C/A99C/K141C/T177C/V199C/N203L as compared to SEQ ID NO: 2, and which further comprises a substitution in one or more position selected from the following: 30, 36, 43, 46, 57, 60, 64, 73, 79, 119, 121, 123, 130, 134, 138, 151, 155, 161, 162, 168, 176, 180, 184, 190, 207, 224, 230, 243, 273, 286, 336, 340, 358 and 375, using SEQ ID NO: 2 for numbering.

2. The variant of claim 1 , comprising substitutions in the positions: 57, 73, 121, 134, 155, 207 and 273.

3. The variant of claim 2 , further comprising substitutions in the positions: 36, 60, 64, 73, 119, 130, 138,161, 162, 168, 176, 180, 184, 190, 224, 230, 243, 336 and 340.

4. The variant of claim 1 , where the substitutions are selected among: 30Q, 36A, 43C, 46C, 57Y, 60H, 64Q, 73P, 79Q, 119P, 121P, 123C, 130T, 130C, 134Q, 138A, 151S, 155F, 161T, 162A, 168R 176P, 180N, 184Q, 190T, 207T, 224Q, 230E, 243N, 273L, 286S, 336R, 340L, 340P, 358Q and 375K.

5. The variant of claim 1 , comprising the substitutions 57Y/73P/121P/134Q/155F/207T/273L.

6. A phytase variant which has phytase activity and an amino acid sequence having at least 70% identity to SEO ID NO: 2, and which comprises alterations and substitutions, as compared to SEO ID NO: 2, selected from the group consisting of:
31C/52C/57Y/73P/99C/121P/134Q/141C/155F/177C/199C/203L/207T/273L;
31C/36A/52C/57Y/60H/64Q/73P/99C/119S/121P/130T/134Q/138A/141C/155F/161T/162A/176P/177C/180N/184Q/190T/199C/203L/207T/224Q/230E/243N/273L/336R/340L;
31C/36A/43C/46C/52C/57Y/60H/64Q/73P/99C/1195/121P/130T/134Q/138A/141C/155F/161T/162A/176P/177C/180N/184Q/190T/199C/203L/207T/224Q/230E/243N/273L/2865/336R/340L;
31C/36A/52C/57Y/60H/64Q/73P/99C/1195/121P/123C/130C/134Q/138A/141C/151S/155F/161T/162A/176P/177C/180N/184Q/190T/199C/203L 207T/224Q/230E/243N/273L/336R/340L; and
31C/36A/43C/46C/52C/57Y/60H/64Q/73P/99C/1195/121P/123C/130C/134Q/138A/141C/155F/161T/162A/176P/177C/180N/184Q/190T/199C/203L/207T/224Q/230E/243N/273L/286S/336R/340L.

7. The variant of claim 6 , having an amino acid sequence which has at least 75% sequence identity to the amino acid sequence of SEQ ID NO: 2 comprising alterations and substitutions, as compared to SEO ID NO: 2, selected from the group consisting of:
N31C/G52C/E57Y/N73P/A99C/N121P/S134Q/K141C/Y155F/T177C/V199C/N203L/P207T/M273L;
N31C/Q36A/G52C/E57Y/Q60H/L64Q/N73P/A99C/E119S/N121P/M13OT/S134Q/L138A/K141C/Y155F/S161T/S162A/E176P/T177C/T180N/S184Q/P190T/V199C/N203L/P207T/E224Q/Q230E/R243N/M273L/K336R/T340L;
N31C/Q36A/P43C/W46C/G52C/E57Y/Q60H/L64Q/N73P/A99C/E119S/N121P/M130T/S134Q/L138A/K141C/Y155F/S161T/S162A/E176P/T177C/T180N/S184Q/P190T/V199C/N203L/P207T/E224Q/Q230E/R243N/M273L/N286S/K336R/T340L;
N31C/Q36A/G52C/E57Y/Q60H/L64Q/N73P/A99C/E119S/N121P/P123C/M130C/S134Q/L138A/K141C/N151S/Y155F/S161T/S162A/E176P/T177C/T180N/S184Q/P190T/V199C/N203L/P207T/E224Q/Q230E/R243N/M273L/K336R/T340L; and
N31C/Q36A/P43C/W46C/G52C/E57Y/Q60H/L64Q/N73P/A99C/E119S/N121P/P123C/M130C/S134Q/L138A/K141C/Y155F/S161T/S162A/E176P/T177C/T180N/S184Q/P190T/V199C/N203L/P207T/E224Q/Q230E/R243N/M273L/N286S/K336R/T340L.

8. A composition comprising at least one phytase variant according to claim 1 .

9. The composition of claim 8 further comprising
at least one fat soluble vitamin;
at least one water soluble vitamin; and/or
at least one trace mineral.

10. The composition of claim 8 , further comprising at least one additional enzyme selected from amylases, phytases other than said phytase variant, phosphatases, xylanases, galactanases, alpha-galactosidases, proteases, phospholipases and beta-glucanases.

11. An animal feed composition having a crude protein content of 50 to 800 g/kg and comprising the phytase variant of claim 1 .

12. The phytase variant of claim 1 , having at least 75% identity to SEQ ID NO: 2.

13. The phytase variant of claim 1 , having at least 80% identity to SEQ ID NO: 2.

14. The phytase variant of claim 1 , having at least 85% identity to SEQ ID NO: 2.

15. The phytase variant of claim 1 , having at least 90% identity to SEQ ID NO: 2.

16. The phytase variant of claim 6 , having at least 80% identity to SEQ ID NO: 2.

17. A phytase variant comprising an amino acid sequence selected from SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18 and SEQ ID NO: 20.

18. The phytase variant of claim 17 , comprising SEQ ID NO: 12.

19. The phytase variant of claim 17 , comprising SEQ ID NO: 14.

20. The phytase variant of claim 17 , comprising SEQ ID NO: 18.

Assignees

• Novozymes As

Inventors

• Zhai Hengxiao
• Vind Jesper
• Skov Lars Kobberoe
• Zhang Qian
• Santigosa Ester
• Sorbara Jose-Otavio B
• Seon Aurelia Anne Catherine Charlotte
• Walk Carrie Louise

Classifications

• C12Y301/03008

  3-Phytase (3.1.3.8) · CPC title

• C12N15/75

  for Bacillus · CPC title

• A23K10/14

  Pretreatment of feeding-stuffs with enzymes · CPC title

• A23K20/30

  Oligoelements · CPC title

• A23K20/189

  Enzymes · CPC title