US 11,746,357 B2
Method for producing HSL protein having improved catalytic activity for 2-oxoglutaric acid-dependently oxidizing 4-HPPD inhibitor
Yuzuru Tozawa, Saitama (JP); Satomi Takei, Saitama (JP); Masahiro Oshima, Tsukuba (JP); Sakiko Hirose, Tsukuba (JP); Motoshige Kawata, Tsu (JP); Keisuke Sekino, Tokyo (JP); and Akihiko Yamazaki, Tokyo (JP)
Assigned to National Agriculture and Food Research Organization, Tsukuba (JP); and SDS BIOTECH K.K., Tokyo (JP)
Appl. No. 16/485,046
Filed by National Agriculture and Food Research Organization, Tsukuba (JP); and SDS Biotech K.K., Tokyo (JP)
PCT Filed Feb. 9, 2018, PCT No. PCT/JP2018/004514
§ 371(c)(1), (2) Date Aug. 9, 2019,
PCT Pub. No. WO2018/147401, PCT Pub. Date Aug. 16, 2018.
Claims priority of application No. 2017-023294 (JP), filed on Feb. 10, 2017.
Prior Publication US 2020/0048315 A1, Feb. 13, 2020
Int. Cl. C12N 15/82 (2006.01); C07K 14/415 (2006.01); C12Q 1/6895 (2018.01)
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.