Phytase enzymes, nucleic acid sequences encoding phytase enzymes and vectors and host cells incorporating same

• US 7,220,445 B2
• Filed: 10/25/2002
• Issued: 05/22/2007
• Est. Priority Date: 10/26/2001
• Status: Expired due to Fees

• Current Assignee: Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Original Assignee: Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Inventors: Goedegebuur, Frits, Power, Scott D., Bower, Benjamin, Clarkson, Kathleen A.
• Primary Examiner(s): Saidha; Tekchand
• Application Number:
  US10/492,782
• Publication Number:
  US 20050069978A1
• Time in Patent Office: 1,670 Days
• Field of Search: 426/656, 435/196, 435/252.3, 435/320.1, 536/23.2
• US Class Current: 426/656
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)

Abstract

DNA is provided which encodes an enzyme having phytase activity isolated from Penicillium, Fusarium, Humicola and Emericella. 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. An isolated polynucleotide encoding an enzyme having phytase activity comprising a nucleotide sequence

(i) having at least 90% identity to the nucleotide sequence of SEQ ID NO:

1 or SEQ ID NO;

43 or (ii) being fully complementary to a nucleotide sequence having at least 90% sequence identity to SEQ ID NO;

1 or SEQ ID NO;

43.

2. An expression construct comprising the polynucleotide of claim 1.

3. A vector including the expression construct of claim 2.

4. An isolated host cell transformed with the vector of claim 3.

7. An isolated phytase enzyme encoded by the polynucleotide of claim 1, wherein said enzyme is obtained from a Penicillium chrysogenum, and has the following physiochemical properties:

(1) molecular weight;

between about 49 and 51 kDa (non-glycosylated); and

(2) substrate;

phytate.

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

(a) providing a host cell transformed with an expression vector comprising a polynucleotide as defined in claim 1;

(b) cultivating said transformed host cell under conditions suitable for said host cell to produce said phytase; and

(c) recovering said phytase.

9. The method of claim 8, wherein said host cell is an Aspergillus species.

10. A purified enzyme having phytase activity, produced by the method of claim 9.

12. A method of separating phosphorous from phytate, comprising:

treating said phytate with an enzyme as defined in claim 8.

15. The method according to claim 8, wherein the polynucleotide encodes an enzyme having phytase activity and at least 90% sequence identity to SEQ ID NO:

2.

5. An isolated polynucleotide encoding an enzyme having phytase activity, wherein said enzyme comprises an amino acid sequence having at least 90% amino acid sequence identity to SEQ ID NO:

2 or SEQ ID NO;

44.

6. Food or animal feed comprising an enzyme having phytase activity, wherein said enzyme comprises an amino acid sequence having at least 90% amino acid sequence identity to SEQ ID NO:

2 or SEQ ID NO;

44.

11. A method of separating phosphorous from phytate, comprising:

treating said phytate with an enzyme having phytase activity comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO;

2 or SEQ ID NO;

44.

13. An isolated polynucleotide encoding an enzyme having phytase activity, wherein said enzyme comprises an amino acid sequence having at least 95% amino acid sequence identity to SEQ ID NO:

2 or SEQ ID NO;

44.

14. Food or animal feed comprising an enzyme having phytase activity, wherein said enzyme comprises an amino acid sequence having at least 95% amino acid sequence identity to SEQ ID NO:

2 or SEQ ID NO;

44.