The present invention provides a fungus of the genus Rhizopus having high productivity of an organic acid. The present invention also provides a mutant of the genus Rhizopus with reduced pyruvate decarboxylase activity.
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1. A mutant of the genus Rhizopus wherein the mutant's pdc gene is deleted or inactivated, wherein the pdc gene is at least one polynucleotide selected from the group consisting of:a polynucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 1;a polynucleotide consisting of a nuc
1. A mutant of the genus Rhizopus wherein the mutant's pdc gene is deleted or inactivated, wherein the pdc gene is at least one polynucleotide selected from the group consisting of:a polynucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 1;a polynucleotide consisting of a nucleotide sequence having at least 80% identity to the nucleotides represented by SEQ ID NO: 1, and encoding a polypeptide which has pyruvate decarboxylase activity;a polynucleotide consisting of a nucleotide sequence having deletion, insertion, substitution or addition of one or more nucleotides with respect to the nucleotide sequence represented by SEQ ID NO: 1, and encoding a polypeptide which has pyruvate decarboxylase activity;a polynucleotide encoding a polypeptide which consists of the amino acid sequence represented by SEQ ID NO: 2;a polynucleotide encoding a polypeptide which consists of an amino acid sequence having at least 80% identity to the amino acid sequence represented by SEQ ID NO: 2 and has pyruvate decarboxylase activity; anda polynucleotide encoding a polypeptide which consists of an amino acid sequence having deletion, insertion, substitution or addition of one or more amino acid residues with respect to the amino acid sequence represented by SEQ ID NO: 2 and has pyruvate decarboxylase activity. 2. The mutant of the genus Rhizopus according to claim 1, wherein the pyruvate decarboxylase activity is reduced to 50% or less as compared with that before mutation. 3. The mutant of the genus Rhizopus according to claim 1, wherein the fungus of the genus Rhizopus is Rhizopus oryzae or Rhizopus delemar. 4. The mutant of the genus Rhizopus according to claim 1, wherein the mutant has improved productivity of an organic acid. 5. The mutant of the genus Rhizopus according to claim 4, wherein the organic acid is fumaric acid, lactic acid, succinic acid, malic acid or α-ketoglutaric acid. 6. A method for producing a mutant of the genus Rhizopus, comprising deleting or inactivating the fungus' pdc gene in a fungus of the genus Rhizopus, whereinthe pdc gene is at least one polynucleotide selected from the group consisting of:a polynucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 1;a polynucleotide consisting of a nucleotide sequence having at least 80% identity to the nucleotides represented by SEQ ID NO: 1, and encoding a polypeptide which has pyruvate decarboxylase activity;a polynucleotide consisting of a nucleotide sequence having deletion, insertion, substitution or addition of one or more nucleotides with respect to the nucleotide sequence represented by SEQ ID NO: 1, and encoding a polypeptide which has pyruvate decarboxylase activity;a polynucleotide encoding a polypeptide which consists of the amino acid sequence represented by SEQ ID NO: 2;a polynucleotide encoding a polypeptide which consists of an amino acid sequence having at least 80% identity to the amino acid sequence represented by SEQ ID NO: 2 and has pyruvate decarboxylase activity; anda polynucleotide encoding a polypeptide which consists of an amino acid sequence having deletion, insertion, substitution or addition of one or more amino acid residues with respect to the amino acid sequence represented by SEQ ID NO: 2 and has pyruvate decarboxylase activity. 7. A method for improving productivity of an organic acid of a fungus of the genus Rhizopus, comprising deleting or inactivating the fungus' pdc gene in the fungus of the genus Rhizopus, whereinthe pdc gene is at least one polynucleotide selected from the group consisting of:a polynucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 1;a polynucleotide consisting of a nucleotide sequence having at least 80% identity to the nucleotides represented by SEQ ID NO: 1, and encoding a polypeptide which has pyruvate decarboxylase activity;a polynucleotide consisting of a nucleotide sequence having deletion, insertion, substitution or addition of one or more nucleotides with respect to the nucleotide sequence represented by SEQ ID NO: 1, and encoding a polypeptide which has pyruvate decarboxylase activity;a polynucleotide encoding a polypeptide which consists of the amino acid sequence represented by SEQ ID NO: 2;a polynucleotide encoding a polypeptide which consists of an amino acid sequence having at least 80% identity to the amino acid sequence represented by SEQ ID NO: 2 and has pyruvate decarboxylase activity; anda polynucleotide encoding a polypeptide which consists of an amino acid sequence having deletion, insertion, substitution or addition of one or more amino acid residues with respect to the amino acid sequence represented by SEQ ID NO: 2 and has pyruvate decarboxylase activity. 8. The method according to claim 6, wherein the deletion or inactivation of the pdc gene is performed by genome editing of a pdc gene locus using a programmable nuclease. 9. The method according to claim 8, wherein the genome editing is performed using TALEN, Crispr-cas9 system, or ZFN. 10. The method according to claim 8, wherein the genome editing is transfer of TALEN peptides or polynucleotides encoding the TALEN peptides to the fungus of the genus Rhizopus. 11. The method according to claim 10, wherein the TALEN peptides consists of the following polypeptides (L) and (R): the polypeptide (L) having a TAL effector having 17 repeats linked, whereinthe 1st to 16th repeats counted from the upstream end among the 17 repeats each consist of an amino acid sequence having at least 85% identity to the amino acid sequence represented by SEQ ID NO: 11,the 17th repeat counted from the upstream end among the 17 repeats consists of an amino acid sequence having at least 95% identity to the amino acid sequence represented by SEQ ID NO: 27, andthe TAL effector recognizes the sequence represented by SEQ ID NO: 9, and the polypeptide (R)having a TAL effector having 17 repeats linked, whereinthe 1st to 16th repeats counted from the upstream end among the 17 repeats each consist of an amino acid sequence having at least 85% identity to the amino acid sequence represented by SEQ ID NO: 28,the 17th repeat counted from the upstream end among the 17 repeats consists of an amino acid sequence having at least 95% identity to the amino acid sequence represented by SEQ ID NO: 44, andthe TAL effector recognizes the sequence represented by SEQ ID NO: 10. 12. The method according to claim 10, wherein the TALEN peptides consists of the following polypeptides (L) and (R): the polypeptide (L) being a polypeptide which consists of the amino acid sequence represented by SEQ ID NO: 4;a polypeptide which consists of an amino acid sequence having at least 95% identity to the amino acid sequence represented by SEQ ID NO: 4 and has a TAL effector targeting the sequence represented by SEQ ID NO: 9 and a DNA cleavage domain consisting of Fok1-like DNA nuclease;a polypeptide which is encoded by a polynucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 3; ora polypeptide which is encoded by a polynucleotide consisting of a nucleotide sequence having at least 95% identity to the nucleotide sequence represented by SEQ ID NO: 3, and has a TAL effector targeting the sequence represented by SEQ ID NO: 9 and a DNA cleavage domain consisting of Fok1-like DNA nuclease, and the polypeptide (R) being a polypeptide which consists of the amino acid sequence represented by SEQ ID NO: 6;a polypeptide which consists of an amino acid sequence having at least 95% identity to the amino acid sequence represented by SEQ ID NO: 6 and has a TAL effector targeting the sequence represented by SEQ ID NO: 10 and a DNA cleavage domain consisting of Fok1-like DNA nuclease;a polypeptide which is encoded by a polynucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 5; ora polypeptide which is encoded by a polynucleotide consisting of a nucleotide sequence having at least 95% identity to the nucleotide sequence represented by SEQ ID NO: 5, and has a TAL effector targeting the sequence represented by SEQ ID NO: 10 and a DNA cleavage domain consisting of Fok1-like DNA nuclease. 13. The method according to claim 10, further comprising transferring an exonuclease or a polynucleotide encoding the exonuclease to the fungus of the genus Rhizopus. 14. The method according to claim 13, wherein the polynucleotide encoding the exonuclease is a polynucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 7;a polynucleotide consisting of a nucleotide sequence having at least 95% identity to the nucleotide sequence represented by SEQ ID NO: 7, and encoding a polypeptide which has exonuclease activity;a polynucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 82;a polynucleotide consisting of a nucleotide sequence having at least 95% identity to the nucleotide sequence represented by SEQ ID NO: 82, and encoding a polypeptide which has exonuclease activity;a polynucleotide encoding a polypeptide which consists of the amino acid sequence represented by SEQ ID NO: 8; ora polynucleotide encoding a polypeptide which consists of an amino acid sequence having at least 95% identity to the amino acid sequence represented by SEQ ID NO: 8 and has exonuclease activity. 15. The method according to claim 6, wherein the fungus of the genus Rhizopus is Rhizopus oryzae or Rhizopus delemar. 16. A method for producing an organic acid, comprising culturing the mutant of the genus Rhizopus according to claim 1. 17. The method for producing an organic acid according to claim 16, further comprising collecting the organic acid from the culture after the culturing. 18. The method for producing an organic acid according to claim 16, wherein the organic acid is fumaric acid, lactic acid, succinic acid, malic acid or α-ketoglutaric acid. 19. The method for producing an organic acid according to claim 16, wherein the culturing is aerobic culturing.
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