Method for manufacturing useful substance

1. A method for producing an objective substance, the method comprising:

culturing a microorganism having an objective substance-producing ability in a medium to produce and accumulate the objective substance in the medium or in cells of the microorganism; and

collecting the objective substance from the medium or the cells,

wherein the microorganism has been modified so that the activity of a dicarboxylic acid exporter protein is reduced;

wherein the gene encoding the dicarboxylic acid exporter protein is selected from the group consisting of yjjP gene, yjjB gene, yeeA gene, ynfM gene, sucE1 gene, and combinations thereof;

wherein the yjjP gene is a DNA selected from the group consisting of:

(A) a DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 158 or 160;

(B) a DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 158 or 160, but including substitution, deletion, insertion, or addition of 1-10 amino acid residues, the protein having an activity to export a dicarboxylic acid;

(C) a DNA comprising the nucleotide sequence of SEQ ID NO: 157 or 159; and

(D) a DNA able to hybridize under stringent conditions with a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 157 or 159, or with a probe that can be prepared from the complementary nucleotide sequence, and encoding a protein having an activity to export a dicarboxylic acid;

wherein the yjjB gene is a DNA selected from the group consisting of:

(A) DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 162 or 164;

(B) DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 162 or 164, but including substitution, deletion, insertion, or addition of 1-10 amino acid residues, the protein having an activity to export a dicarboxylic acid;

(C) DNA comprising the nucleotide sequence of SEQ ID NO: 161 or 163; and

(D) DNA able to hybridize under stringent conditions with a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 161 or 163, or with a probe that can be prepared from the complementary nucleotide sequence, and encoding a protein having an activity to export a dicarboxylic acid;

wherein the yeeA gene is a DNA selected from the group consisting of:

(A) DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 166, 168, or 170;

(B) DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 166, 168, or 170, but including substitution, deletion, insertion, or addition of 1-10 amino acid residues, the protein having an activity to export a dicarboxylic acid;

(C) DNA comprising the nucleotide sequence of SEQ ID NO: 165, 167, or 169; and

(D) DNA able to hybridize under stringent conditions with a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 165, 167, or 169, or with a probe that can be prepared from the complementary nucleotide sequence, and encoding a protein having an activity to export a dicarboxylic acid;

wherein the ynfM gene is a DNA selected from the group consisting of:

(A) DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 172, 174, 176, 178, or 180;

(B) DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 172, 174, 176, 178, or 180, but including substitution, deletion, insertion, or addition of 1-10 amino acid residues, the protein having an activity to export a dicarboxylic acid;

(C) DNA comprising the nucleotide sequence of SEQ ID NO: 171, 173, 175, 177, or 179; and

(D) DNA able to hybridize under stringent conditions with a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 171, 173, 175, 177, or 179, or with a probe that can be prepared from the complementary nucleotide sequence, and encoding a protein having an activity to export a dicarboxylic acid;

wherein the sucE1 gene is a DNA selected from the group consisting of:

(A) DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 278 or 280;

(B) DNA encoding a protein comprising the amino acid sequence of SEQ ID NO: 278 or 280, but including substitution, deletion, insertion, or addition of 1-10 amino acid residues, the protein having an activity to export a dicarboxylic acid;

(C) DNA comprising the nucleotide sequence of SEQ ID NO: 277 or 279; and

(D) DNA able to hybridize under stringent conditions with a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 277 or 279, or with a probe that can be prepared from the complementary nucleotide sequence, and encoding a protein having an activity to export a dicarboxylic acid;

wherein said stringent conditions are 0.1×SSC, 0.1% SDS at 68° C.; and

wherein the objective substance is a metabolite derived from acetyl-CoA and/or an L-amino acid.

2. The method according to claim 1 , wherein the activity of the dicarboxylic acid exporter protein is reduced by attenuating the expression of a gene encoding the dicarboxylic acid exporter protein or by deleting the gene.

3. The method according to claim 1 , wherein the metabolite derived from acetyl-CoA and/or the L-amino acid is selected from the group consisting of isopropyl alcohol, ethanol, acetone, propylene, isoprene, 1,3-butanediol, 1,4-butanediol, 1-propanol, 1,3-propanediol, 1,2-propanediol, ethylene glycol, isobutanol, and combinations thereof.

4. The method according to claim 1 , wherein the metabolite derived from acetyl-CoA and/or the L-amino acid is selected from the group consisting of citric acid, itaconic acid, acetic acid, butyric acid, 3-hydroxybutyric acid, polyhydroxybutyric acid, 3-hydroxyisobutyric acid, 3-aminoisobutyric acid, 2-hydroxyisobutyric acid, methacrylic acid, 6-aminocaproic acid, and combinations thereof.

5. The method according to claim 1 , wherein the metabolite derived from acetyl-CoA and/or the L-amino acid is selected from the group consisting of polyglutamic acid, L-glutamic acid, L-glutamine, L-arginine, L-ornithine, L-citrulline, L-leucine, L-isoleucine, L-valine, L-cysteine, L-serine, L-proline, and combinations thereof.

6. The method according to claim 5 , wherein the L-glutamic acid is monoammonium L-glutamate or monosodium L-glutamate.

7. The method according to claim 1 , wherein the microorganism is a coryneform bacterium or a bacterium belonging to the family Enterobacteriaceae.

8. The method according to claim 7 , wherein the coryneform bacterium is Corynebacterium glutamicum.

9. The method according to claim 7 , wherein the bacterium belonging to the family Enterobacteriaceae is Escherichia coli, Pantoea ananatis , or Enterobacter aerogenes.

10. The method according to claim 1 , wherein the dicarboxylic acid is selected from the group consisting of malic acid, succinic acid, fumaric acid, 2-hydroxyglutaric acid, and α-ketoglutaric acid.

11. The method according to claim 1 , wherein the microorganism has been further modified so that malyl-CoA-producing ability is increased.

12. The method according to claim 1 , wherein the microorganism has been further modified so that α-ketoglutarate synthase activity is increased.