CPC C12N 15/8274 (2013.01) [C07K 14/415 (2013.01); C12Q 1/6895 (2013.01)] | 2 Claims |
1. A method for producing a mutant HSL (4-Hydroxyphenylpyruvate dioxygenase Inhibitor sensitive 1-Like) protein with increased catalytic activity to oxidize a 4-HPPD (4-hydroxyphenylpyruvate dioxygenase) inhibitor in a 2-oxoglutarate-dependent manner,
said method comprising:
expressing, in a plant cell, a mutant HSL protein, wherein said mutant HSL protein is a mutant of a wild type HSL protein, wherein said wild type HSL protein comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence of any one of SEQ ID NO: 4, 18, 20, or 22,
wherein the mutant HSL protein comprises the substitution of a phenylalanine or glutamine to a basic amino acid at a position corresponding to position 140 of SEQ ID NO: 4, and optionally further comprises one or both of the following substitutions: (1) a leucine or tyrosine to phenylalanine at a position corresponding to position 204 of SEQ ID NO: 4, and (2) a phenylalanine to leucine at a position corresponding to position 298 of SEQ ID NO: 4, wherein the basic amino acid is lysine, histidine or arginine, and
wherein said mutant HSL protein has no more than three substitutions relative to the wild type HSL protein.
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