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US 9,422,533 B2Granted
Nucleic acid encoding N-methylputrescine oxidase and uses thereof
Assignee22nd Century Limited, LLC
InventorsJonathan E. Page, Enwu Liu
Life of this patent
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Abstract
The gene encoding N-methylputrescine oxidase (MPO) and constructs comprising such DNA are provided, including methods of regulating MPO expression independently or with other alkaloid biosynthesis genes to modulate alkaloid production in plants and host cells. MPO genes or fragments thereof are useful for reducing pyrrolidine or tropane alkaloid production in plants, for increasing pyrrolidine or tropane alkaloid production in plants, and for producing an MPO enzyme in host cells.
Claims15
A method of producing MPO enzyme, comprising
- (1) genetically engineering a cell with a nucleic acid construct comprising an isolated nucleic acid molecule selected from the group consisting of:
- (a) the nucleotide sequence set forth in SEQ ID NO:
- 1;
- (b) the nucleotide sequence set forth in SEQ ID NO:
- 2;
- (c) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO:
- 3;
- (d) a nucleotide sequence that is at least 95% identical to the full length nucleotide sequences of (a), (b), or (c) and encodes an N-methylputrescine oxidase (MPO) enzyme;
- (e) a nucleotide sequence that hybridizes under high stringent conditions to the full length nucleotide sequences of (a), (b), (c), or (d) and encodes an MPO enzyme;
- and
- (f) a nucleotide sequence that differs from the nucleic acid sequence of (a) or (b) due to the degeneracy of the genetic code and encodes an MPO enzyme;
- and
- (2) growing said engineered cell under conditions such that MPO is produced.
10 more elements
A recombinant MPO enzyme having the amino acid sequence set forth in SEQ ID NO:
- 3, or a variant thereof, wherein the MPO enzyme is fused to a polyhistidine tag.
Dependencies are inferred from the claim text.