Variant E1 protein gene for pyruvate dehydrogenase complex and variant E1 protein of pyruvate dehydrogenase complex

• US 5,432,071 A
• Filed: 03/22/1994
• Issued: 07/11/1995
• Est. Priority Date: 04/08/1993
• Status: Expired due to Fees

• Current Assignee: Kikkoman Corporation
• Original Assignee: Kikkoman Corporation
• Inventors: Otake, Hideko, Koyama, Yasuji, Ichikawa, Toshio, Nakano, Eiichi
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Moore, William W.
• Application Number:
  US08/215,709
• Time in Patent Office: 476 Days
• Field of Search: 435/191, 435/189, 435/190, 435/320.1, 435/69.1, 435/252.3, 435/252.33, 435/, 536/23.2
• US Class Current: 435/190
• CPC Class Codes: C12N 9/0008 acting on the aldehyde or oxo group of donors (1.2)
• CPC Class Codes: C12Y 102/04001 Pyruvate dehydrogenase (acetyl-transferring) (1.2.4.1)
• CPC Class Codes: C12Y 102/05001 Pyruvate dehydrogenase (quinone) (1.2.5.1)

Abstract

The present invention relates to variant E₁ protein of pyruvate dehydrogenase complex of high activity in which arginine at the 146-position is replaced by proline in the amino acid sequence of wild-type E₁ protein of pyruvate dehydrogenase complex, a gene coding for said variant E₁ protein, a novel recombinant DNA comprising said variant E₁ protein-encoding gene inserted in a vector DNA, and a process for producing variant E₁ protein by said recombinant DNA. The present invention provides the variant E₁ protein of pyruvate dehydrogenase complex of high activity.

Claims

1. An isolated gene coding for the amino acid sequence of variant E₁ protein of E. coli pyruvate dehydrogenase complex in which arginine at the 146-position is replaced by proline in the amino acid sequence of wild-type E₁ protein of E. coli pyruvate dehydrogenase complex.

2. A recombinant DNA comprising the gene for the variant E₁ protein of E. coli pyruvate dehydrogenase complex of claim 1 inserted into a vector DNA.

3. A process for producing variant E₁ protein of E. coli pyruvate dehydrogenase complex, comprising culturing a microorganism of genus Escherichia carrying the recombinant DNA of claim 2 and recovering the variant E₁ protein of E. coli pyruvate dehydrogenase complex from the culture.

4. Variant E₁ protein of E. coli pyruvate dehydrogenase complex in which arginine at the 146-position is replaced by proline in the amino acid sequence of wild-type E₁ protein of E. coli pyruvate dehydrogenase.