Compositions and methods of zearalenone detoxification

A plant cell having stably incorporated in its genome a nucleotide sequence operably linked to a promoter active in said cell, wherein said nucleotide sequence is selected from the group consisting of:

• a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• b) a nucleotide sequence comprising at least 40 contiguous nucleotides of the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• c) a nucleotide sequence comprising the cDNA insert of the plasmid deposited as Patent Deposit No. ______, ______, or ______;
• d) a nucleotide sequence comprising a sequence having at least 70% identity to the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5, wherein said sequence encodes a polypeptide having zearalenone detoxifying activity;
• e) a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:7;
• and,
• f) a nucleotide sequence hybridizing under stringent conditions to the complement of a nucleotide sequence of a), b), c), d), or e), wherein said sequence encodes a polypeptide having zearalenone detoxification activity.

A plant having stably incorporated in its genome a nucleotide sequence operably linked to a promoter active in said plant, wherein said nucleotide sequence is selected from the group consisting of:

• a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• b) a nucleotide sequence comprising at least 40 contiguous nucleotides of the sequences set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• c) a nucleotide sequence comprising the cDNA insert of the plasmid deposited as Patent Deposit No. ______, ______, or ______;
• d) a nucleotide sequence comprising a sequence having at least 70% identity to the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5, wherein said sequence encodes a polypeptide having a zearalenone detoxification activity;
• e) a nucleotide sequence encoding the amino acid sequences of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:7;
• and,
• f) a nucleotide sequence hybridizing under stringent conditions to a complement of a sequence of a), b), c), d) or e), wherein said sequence encodes a polypeptide having zearalenone detoxification activity.

A microorganism having stably incorporated a heterologous nucleotide sequence operably linked to a promoter active in said microorganism, wherein said heterologous nucleotide sequence is selected from the group consisting of:

• a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• b) a nucleotide sequence comprising at least 40 contiguous nucleotides of the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• c) a nucleotide sequence comprising the cDNA insert of the plasmid deposited as Patent Deposit No. ______, ______, or ______;
• d) a nucleotide sequence comprising a sequence having at least 70% identity to the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5, wherein said sequence encodes a polypeptide having a zearalenone detoxification activity;
• e) a nucleotide sequence encoding the amino acid sequences of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:7;
• and,
• f) a nucleotide sequence hybridizing under stringent conditions to a complement of a sequence of a), b), c), d) or e), wherein said sequence encodes a polypeptide that retains zearalenone detoxification activity.

An isolated polypeptide comprising an amino acid sequence selected from the group consisting of:

• a) a polypeptide encoded by the nucleotide sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• b) a polypeptide comprising the amino acid sequences set forth in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:7;
• c) a polypeptide sequence comprising at least 40 contiguous amino acids of the sequence set forth in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:7;
• d) a polypeptide encoded by the nucleic acid sequence of the cDNA insert of the plasmid deposited as Patent Deposit No. ______, ______, or ______;
• and,
• e) a polypeptide having at least 60% identity to the sequence set forth in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:7, or a fragment thereof, wherein said polypeptide retains zearalenone detoxification activity.

A method for detoxifying zearalenone in a plant, said method comprising stably incorporating into a plant cell a nucleotide sequence operably linked to a promoter that is capable of initiating transcription in a plant cell, wherein said nucleotide sequence comprises a nucleotide sequence selected from the group consisting of:

• a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• b) a nucleotide sequence comprising at least 40 contiguous nucleotides of the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• c) a nucleotide sequence comprising the cDNA insert of the plasmid deposited as Patent Deposit No. ______, ______, or ______;
• d) a nucleotide sequence comprising a sequence having at least 70% identity to the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5, wherein said sequence encodes a polypeptide having a zearalenone detoxification activity;
• e) a nucleotide sequence encoding the amino acid sequences of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:7;
• and,
• f) a nucleotide sequence hybridizing under stringent conditions to a complement of a sequence of a), b), c), d) or e), wherein said sequence encodes a polypeptide that retains zearalenone detoxification activity.

A method for the detoxification of zearalenone, said method comprising contacting said zearalenone with microorganisms expressing a heterologous nucleotide sequence selected from the group consisting of:

• a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• b) a nucleotide sequence comprising at least 40 contiguous nucleotides of the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• c) a nucleotide sequence comprising the cDNA insert of the plasmid deposited as Patent Deposit No. ______, ______, or ______;
• d) a nucleotide sequence comprising a sequence having at least 70% identity to the sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ i) NO:5, wherein said sequence encodes a polypeptide having zearalenone detoxification activity;
• e) a nucleotide sequence encoding the amino acid sequences of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:7;
• and,
• f) a nucleotide sequence hybridizing under stringent conditions to a complement of a sequence of a), b), c), d) or e), wherein said sequence encodes a polypeptide having zearalenone detoxification activity.

A method for the detoxification of zearalenone, said method comprising contacting said zearalenone with a polypeptide selected from the group consisting of:

• a) a polypeptide encoded by the nucleotide sequence set forth in SEQ ID NO:1, SEQ ID NO:3, or SEQ ID NO:5;
• b) a polypeptide comprising the amino acid sequences set forth in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:7;
• c) a polypeptide sequence comprising at least 40 contiguous amino acids of the sequence set forth in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:7;
• d) a polypeptide encoded by the nucleic acid sequence of the cDNA insert of the plasmid deposited as Patent Deposit No. ______, ______, or ______;
• e) a polypeptide having at least 60% identity to the sequence set forth in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:7, or a fragment thereof, wherein said polypeptide retains zearalenone detoxification activity.

A method for the expression of a polypeptide comprising:

• a) stably introducing into an isolated non-human animal cell an expression cassette comprising a nucleotide sequence, wherein said nucleotide sequence is selected from the group consisting of:
• i) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:1, 3, or 5;
• ii) a nucleotide sequence comprising at least 40 contiguous nucleotides of the sequence set forth in SEQ ID NO:1, 3, or 5;
• iii) a nucleotide sequence comprising the cDNA insert of the plasmid deposited as Patent Deposit No. ______, ______, or ______;
• iv) a nucleotide sequence comprising a sequence having at least 70% identity to the sequence set forth in SEQ ID NO:1, 3, or 5, wherein said sequence encodes a polypeptide having zearalenone, and structural analogs thereof, detoxifying activity;
• v) a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:2, 4, 6, or 7;
• and
• vi) a nucleotide sequence that hybridizes under stringent conditions to a sequence of i), ii), iii), iv), v), or a complement thereof;
• b) culturing said non-human animal cell under conditions such that the polypeptide encoded by the nucleotide sequence is expressed in said non-human animal cell.