IP Library Granted Patent US 7,078,183
Granted Patent B2
US 7,078,183 · App. 10/776,104 · Granted Jul 18, 2006

Modified phytases

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Quick Facts
Patent No.
US 7,078,183
App. No.
10/776,104
Granted
Jul 18, 2006
Kind
B2
Abstract

A process for the production of a modified phytase with a desired property improved over the property of the corresponding unmodified phytase is disclosed, as well as modified phytases, polynucleotides encoding modified phytases, and animal feed including modified phytases.

Claims (14)

1. A process for preparing a modified Aspergillus phytase with a specific activity improved over the specific activity of an unmodified Aspergillus phytase which comprises:

(a) determining the three dimensional structure of the unmodified Aspergillus phytase and of a second phytase which has the improved specific activity by aligning the amino acid sequences of said phytases with the amino acid sequence of a third phytase which is the phytase of Aspergillus niger and using the three dimensional structure of the phytase of Aspergillus niger as a template based on the alignment to determine said three dimensional structures;

(b) determining from the structures of step (a) the amino acids of the active sites of the unmodified Aspergillus phytase and of the second phytase having the improved specific activity which active site provides the improved specific activity and comparing the amino acids which form the active sites to identify which amino acids are different in the active site of the second phytase from the amino acids in the active site of the unmodified Aspergillus phytase;

(c) constructing a DNA sequence coding for the modified phytase by obtaining the DNA sequence of the unmodified Aspergillus phytase and changing the nucleotides coding for the active site which provides the improved specific activity for said unmodified Aspergillus phytase so that at least one of the amino acids in the active site which provides the improved specific activity is substituted by one of the amino acids which was identified as being different in step (b);

(d) integrating such a DNA sequence into a vector capable of expression in a suitable host cell; and

(e) transforming the suitable host cell by the DNA sequence of step (c) or the vector of step (d), growing said host cell under suitable growth conditions and isolating the modified phytase from the host cell or the culture medium.

2. The process of claim 1 wherein the unmodified phytase is a phytase from Aspergillus fumigatus.

3. The process of claim 1 wherein the phytase with the improved specific activity is of eukaryotic origin.

4. The process of claim 3 wherein the phytase with the improved specific activity is of fungal origin.

5. The process of claim 4 wherein the phytase with the improved specific activity is of Aspergillus origin.

6. The process of claim 5 wherein the phytase with the improved specific activity is a phytase from Aspergillus terreus.

7. The process of claim 1 wherein the unmodified phytase is a phytase of Aspergillus fumigatus and the second phytase with the improved specific activity is an the Aspergillus niger phytase.

8. The process of claim 1 wherein the unmodified phytase is a phytase of Aspergillus fumigatus and the second phytase with the improved specific activity is an the Aspergillus terreus phytase.

9. The process according to claim 1 , wherein the DNA sequence encoding the modified Aspergillus phytase has been changed at a position corresponding to position 27 of the phytase of Aspergillus niger (SEQ ID NO:1) to an amino acid selected from the group consisting of Ala, Val, Leu, lie, Thr, Gly, and Asn.