Trichoderma reesei phytase enzymes, nucleic acids encoding such phytase enzymes, vectors and host cells incorporating same and methods of making and using same

• US 7,510,831 B2
• Filed: 10/09/2002
• Issued: 03/31/2009
• Est. Priority Date: 10/26/2001
• Status: Expired due to Term

• Current Assignee: Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Original Assignee: Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Inventors: Ward, Michael, Goedegebuur, Frits, Dunn-Coleman, Nigel
• Primary Examiner(s): Guzo; David
• Assistant Examiner(s): Joike; Michele K
• Application Number:
  US10/492,783
• Publication Number:
  US 20050130148A1
• Time in Patent Office: 2,365 Days
• Field of Search: None
• US Class Current: 435/6.13
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)

Abstract

A novel DNA is provided which encodes an enzyme having phytase activity isolated from Trichoderma. Also provided for is a method of isolating DNA encoding an enzyme having phytase activity from organisms which possess such DNA, transformation of the DNA into a suitable host organism, expression of the transformed DNA and the use of the expressed phytase protein in feed as a supplement.

Claims

1. A method of producing an enzyme having phytase activity, comprising:

(a) providing a host cell transformed with an expression vector comprising a polynucleotide which codes for a enzyme having phytase activity having the amino acid sequence with at least 90% sequence identity over the entire length of SEQ ID NO;

5, wherein the phosphate binding region has a sequence (V/L)L(A/S)RHGAR;

(b) culturing said transformed host cell under culture conditions for production of said enzyme; and

(c) recovering said enzyme wherein said phytase hydrolyzes phytate to inositol in a phytase assay.

2. The method of claim 1, wherein said host cell is an Aspergillus species.

3. The method of claim 1, wherein said host cell is a Trichoderma species.

4. A purified enzyme having phytase activity, produced by the method of claim 1, said enzyme having an amino acid sequence with at least 90% sequence identity to SEQ ID NO:

5, wherein the phosphate binding region has a sequence (V/L)L(A/S)RHGAR.

5. The method according to claim 1, wherein the polynucleotide codes for an enzyme having phytase activity having at least 95% sequence identity to SEQ ID NO:

5.

6. The method according to claim 1, wherein the polynucleotide has at least 85% sequence identity to the nucleotide sequence disclosed in SEQ ID NOS:

1, 2 or 3.

7. An enzyme composition comprising the purified enzyme of claim 4.

8. The method of claim 2, wherein the Aspergillus species is A. niger.

9. The method of claim 3, wherein the Trichoderma species is T. reesei.

10. The method according to claim 5, wherein the polynucleotide codes for an enzyme comprising SEQ ID NO:

5.

11. An enzyme composition comprising the recovered enzyme of claim 1.

12. The method of claim 1, wherein at least one of the domains selected from the group consisting of:

Box 2, Box 4, box 5, and Box 6 in SEQ ID NO;

5 is conserved.