Feedback-resistant pyruvate carboxylase gene from corynebacterium
AssigneeArcher Daniels Midland Company
InventorPaul D. Hanke
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Abstract
The present invention relates to a mutated pyruvate carboxylase gene from Corynebacterium. The mutant pyruvate carboxylase gene encodes a pyruvate carboxylase enzyme which is resistant to feedback inhibition from aspartic acid. The present invention also relates to a method of replacing the wild-type pyruvate carboxylase gene in Corynebacterium with this feedback-resistant pyruvate carboxylase gene. The present invention further relates to methods of the production of amino acids, preferably lysine, comprising the use of this mutant pyruvate carboxylase enzyme in microorganisms.
Claims20
An isolated or purified nucleic acid molecule comprising a nucleotide sequence which codes for a pyruvate carboxylase enzyme of SEQ ID NO:19, wherein said pyruvate carboxylase enzyme contains at least one mutation which desensitizes said pyruvate carboxylase enzyme to feedback inhibition by aspartic acid selected from the group consisting of:
- a) methionine at position 1 is replaced with a valine,
- b) glutamic acid at position 153 is replaced with an aspartic acid,
- c) alanine at position 182 is replaced with a serine,
- d) alanine at position 206 is replaced with a serine,
- e) histidine at position 227 is replaced with an arginine,
- f) alanine at position 452 is replaced with a glycine, and
- g) aspartic acid at position 1120 is replaced with a glutamic acid.
An isolated or purified nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of:
- a) the nucleotide sequence encoding amino acids 1 to 1157 of SEQ ID NO:2;
- b) the nucleotide sequence encoding amino acids 1 to 1140 of SEQ ID NO:4;
- c) a nucleotide sequence encoding the amino acid sequence encoded by the DNA contained in Deposit Number NRRL B-30293;
- and
- d) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b) or (c).
An isolated or purified polypeptide comprising the amino acid sequence of the polypeptide encoded by the DNA plasmid encoding pyruvate carboxylase contained in Deposit Number NRRL B-11474, the amino acid sequence of SEQ ID NO:2 or the amino acid sequence of SEQ ID NO:4.
An isolated or purified polypeptide comprising an amino acid sequence selected from the group consisting of:
- SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16 and SEQ ID NO:18.
Dependencies are inferred from the claim text.