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US7416874: Recombinant bacterial phytases and uses thereof

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Filing Information

Inventor(s) Jay M. Short ·
Assignee(s) Verenium Corporation ·
Attorney/Agent(s) Morrison & Foerster LLP ·
Primary Examiner Delia M Ramirez ·
Application Number US10430356
Filing date 05/05/2003
Issue date 08/26/2008
Prior Publication Data
Predicted expiration date 08/13/2017
U.S. Classifications 435/196  · 435/252.3  · 435/691  · 435/6  · 435/440  · 435/348  · 435/419  · 435/325  · 435/254.11  · 435/254.2  · 530/350  · 536/232  · 800/278  · 435/320.1  ·
International Classifications A01H106  · C07H2100  · C07K1400  · C12N115  · C12N119  · C12N121  · C12N1500  · C12N506  · C12N510  · C12N916  · C12P2100  · C12Q168  ·
Kind CodeB2
Related U.S. Application DataCROSS-REFERENCE TO RELATED APPLICATIONS
This application is a divisional and claims the benefit of priority under 35 U.S.C. §120 to of U.S. patent application Ser. No. U.S. Ser. No. 09/580,515, filed May 25, 2000, now U.S. Pat. No. 6,720,014, issued Apr. 13, 2004, and divisional U.S. Ser. No. 10/034,985, filed Dec. 21, 2001, which is a continuation-in-part of U.S. Ser. No. 09/318,528, filed May 25, 1999, now U.S. Pat. No. 6,183,740, which is a continuation-in-part of U.S. Ser. No. 09/291,931, filed Apr. 13, 1999, now U.S. Pat. No. 6,190,897, which is a continuation of U.S. Ser. No. 09/259,214, filed Mar. 1, 1999, now U.S. Pat. No. 6,110,719, which is a divisional of U.S. Ser. No. 08/910,798, now U.S. Pat. No. 5,876,997, filed Aug. 13, 1997, the contents of which are incorporated by reference in their entirety herein.
68 Claims, 3 Drawings


Abstract

A purified recombinant phytase enzyme derived from Escherichia coli B. The enzyme has a molecular weight of about 47.1 kilodaltons and has phytase activity. The enzyme can be produced from native or recombinant host cells and can be used to aid in the digestion of phytate where desired. In particular, the phytase of the present invention can be used in foodstuffs to improve the feeding value of phytate rich ingredients.

Independent Claims | See all claims (68)

  1. 1. An isolated, synthetic or recombinant nucleic acid encoding a polypeptide having phytase activity that is expressed and secreted in a yeast cell, wherein the nucleic acid is generated by a method comprising the following steps: (a) providing a nucleic acid comprising (i) the sequence of SEQ ID NO: 1, (ii) the sequence of SEQ ID NO: 1, wherein T can also be U, (iii) the sequence of a nucleic acid encoding a polypeptide comprising the sequence of SEQ ID NO: 2, (iv) the sequence of a nucleic acid encoding a polypeptide having the amino acid sequence of SEQ ID NO: 2 from amino acid residues 1 to 432, (v) the sequence of a nucleic acid encoding a polypeptide having the amino acid sequence of SEQ ID NO: 2 from amino acid residue 1 to 432 and a heterologous amino acid sequence; (vi) the sequence of SEQ ID NO: 1 from nucleotide residue 1 to 1296, or (vii) sequences completely complementary to (i), (ii), (iii), (iv), (v), or (vi); (b) modifying one or more nucleotides in the nucleic acid to another nucleotide, deleting one or more nucleotides in the nucleic acid, or adding one or more nucleotides to the nucleic acid, wherein the modified nucleic acid encodes a polypeptide having phytase activity and the modified nucleic acid hybridizes under stringent hybridization conditions to the nucleic acid of SEQ ID NO: 1, wherein the hybridization occurs because the modified nucleic acid has at least 95% sequence identity to SEQ ID NO: 1, and all the modifications result in a conservative amino acid substitution of one amino acid for another amino acid; and (c) adding to the nucleic acid of (b) a nucleic acid encoding a yeast secretory signal peptide or transit peptide; wherein expressing the nucleic acid in a yeast cell generates a secreted polypeptide having phytase activity.
  2. 36. An isolated, synthetic or recombinant nucleic acid encoding a polypeptide having phytase activity generated by a method comprising the following steps: (a) providing a nucleic acid comprising (i) the sequence of SEQ ID NO: 1, (ii) the sequence of SEQ ID NO: 1, wherein T can also be U, (iii) the sequence of a nucleic acid encoding a polypeptide comprising the sequence of SEQ ID NO: 2, (iv) the sequence of a nucleic acid encoding a polypeptide having the amino acid sequence of SEQ ID NO: 2 from amino acid residues 1 to 432, (v) the sequence of a nucleic acid encoding a polypeptide having the amino acid sequence of SEQ ID NO: 2 from amino acid residue 1 to 432 and a heterologous amino acid sequence; (vi) the sequence of SEQ ID NO: 1 from nucleotide residue 1 to 1296, or (vii) sequences completely complementary to (i), (ii), (iii), (iv), (v), or (vi); (b) modifying one or more nucleotides in the nucleic acid to another nucleotide, deleting one or more nucleotides in the nucleic acid, or adding one or more nucleotides to the nucleic acid, wherein the modified nucleic acid encodes a polypeptide having phytase activity and the modified nucleic acid hybridizes under stringent hybridization conditions to the nucleic acid of SEQ ID NO: 1, wherein the hybridization occurs because the modified nucleic acid has at least 95% sequence identity to SEQ ID NO: 1, and all the modifications result in a conservative amino acid substitution of one amino acid for another amino acid; and (c) adding to the nucleic acid of (b) a nucleic acid encoding a heterologous secretory signal peptide or transit peptide; wherein expressing the nucleic acid produces a polypeptide having phytase activity.
  3. 42. An isolated, synthetic or recombinant nucleic acid encoding a polypeptide having phytase activity that is expressed in a plant cell, wherein the nucleic acid is generated by a method comprising the following steps: (a) providing a nucleic acid comprising (i) the sequence of SEQ ID NO: 1, (ii) the sequence of SEQ ID NO: 1, wherein T can also be U, (iii) the sequence of a nucleic acid encoding a polypeptide comprising the sequence of SEQ ID NO: 2, (iv) the sequence of a nucleic acid encoding a polypeptide having the amino acid sequence of SEQ ID NO: 2 from amino acid residues 1 to 432, (v) the sequence of a nucleic acid encoding a polypeptide having the amino acid sequence of SEQ ID NO: 2 from amino acid residue 1 to 432 and a heterologous amino acid sequence; (vi) the sequence of SEQ ID NO: 1 from nucleotide residue 1 to 1296, or (vii) sequences completely complementary to (i), (ii), (iii), (iv), (v), or (vi); (b) modifying one or more nucleotides in the nucleic acid to another nucleotide, deleting one or more nucleotides in the nucleic acid, or adding one or more nucleotides to the nucleic acid, wherein the modified nucleic acid encodes a polypeptide having phytase activity and the modified nucleic acid hybridizes under stringent hybridization conditions to the nucleic acid of SEQ ID NO: 1, wherein the hybridization occurs because the modified nucleic acid has at least 95% sequence identity to SEQ ID NO: 1, and all the modifications result in a conservative amino acid substitution of one amino acid for another amino acid; and (c) adding to the nucleic acid of (b) a nucleic acid encoding a plant secretory signal peptide or transit peptide; wherein expressing the nucleic acid in a plant cell produces a polypeptide having phytase activity.

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Referenced By

The current document is not referenced by other documents.

Patent Family

Document NumberAssigneeInventorsIssue/Pub Date
US20030232041 Keith Kretz et al. Dec 2003
US7416874 Verenium Corporation Jay M. Short Aug 2008