Lactonase and stabilized mutants thereof for treating fungal infections in plants
AssigneeMigal Galilee Research Institute Ltd.
InventorsLivnat AFRIAT-JURNOU, Dov PRUSKY
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Abstract
Methods for treating or preventing infection of a fungus secreting patulin in plants or products made therefrom; and for reducing the concentration of patulin in plants, products made therefrom, or non-plant food products, using a lactonase such as an acyl-homoserine lactonase, e.g., a phosphotriesterase-like lactonase; or the wild-type putative parathion hydrolase from M. tuberclorosis (PPH) or a mutant thereof, or a functional fragment thereof.
Claims23
A method for treating or preventing infection of a fungus in a plant or a part, organ or a propagation material thereof, or in a product made from said plant, part, organ or propagation material, said fungus secreting patulin, and said method comprising applying a lactonase or a functional fragment thereof on said plant, part, organ or propagation material;
- or to said product, wherein said lactonase is (i) an acyl-homoserine lactonase;
- or (ii) the wild-type putative parathion hydrolase from M. tuberclorosis (PPH;
- SEQ ID NO:
- 1), or a phosphotriesterase-like lactonase having at least 30% identity to said wild-type PPH, a TIM-barrel fold substantially identical to that of the wild-type PPH, and preserved catalytic residues in its active site.
The method of claim 2, wherein said phosphotriesterase-like lactonase is a mutated phosphotriesterase-like lactonase in which the glycine residue corresponding to G59 of SEQ ID NO:
- 1 is substituted by valine or the histidine residue corresponding to H172 of SEQ ID NO:
- 1 is substituted by tyrosine;
- and said mutated phosphotriesterase-like lactonase has an increased thermostability in comparison with thermostability of a non-mutated wild-type phosphotriesterase-like lactonase or substantially similar or higher lactonase catalytic activity provided with N-(3-oxo-hexanoyl)-homoserine lactone as a substrate in comparison with said non-mutated phosphotriesterase-like lactonase. The method of claim 9, wherein said mutated phosphotriesterase-like lactonase comprises or essentially consists of the amino acid sequence as set forth in SEQ ID NO:
- 2 or SEQ ID NO:
- 3;
- said increased thermostability expressed as T50 is about 55° C. to about 80° C.;
- and/or said mutated phosphotriesterase-like lactonase has an extended shelf-life as compared with said non-mutated phosphotriesterase-like lactonase.
The method of claim 15, wherein said phosphotriesterase-like lactonase is a mutated phosphotriesterase-like lactonase, in which a glycine residue corresponding to G59 of SEQ ID NO:
- 1 is substituted by an amino acid residue selected from the group consisting of valine, alanine, leucine, and isoleucine, or a histidine residue corresponding to H172 of SEQ ID NO:
- 1 is substituted by an amino acid residue selected from the group consisting of tyrosine, phenylalanine and tryptophan.
The method of claim 16, wherein the histidine residue corresponding to H172 of SEQ ID NO:
- 1 is substituted by tyrosine. The method of claim 19, wherein said mutated phosphotriesterase-like lactonase comprises or essentially consists of the amino acid sequence as set forth in SEQ ID NO:
- 3.
The method of claim 15, wherein said acyl-homoserine lactonase is the wild-type putative acyl-homoserine lactonase from E. amylovora (EaAiiA;
- SEQ ID NO:
- 7), or an acyl-homoserine lactonase having at least 30% identity to said wild-type EaAiiA, a MBL fold substantially identical to that of the wild-type EaAiiA, and preserved catalytic residues in its active site.
The method of claim 15, wherein said plant is selected from the group consisting of apple tree, cherry tree, blueberry shrub, plum tree, banana tree, strawberry bush, grape vine, barley grain, wheat grain, and corn grain;
- said organ is a fruit of said plant;
- said plant product is a sauce, juice, jam, or an alcoholic beverage, made from said fruit;
- and said non-plant food product is shellfish.
Dependencies are inferred from the claim text.