3.1.1.45 A150T site-directed mutagenesis, the mutant shows slightly higher Vmax with olmesartan medoxomil compared to the wild-type enzyme 729629 3.1.1.45 C123A inactive 94355 3.1.1.45 C123S - 94357 3.1.1.45 C123S 100% activity towards alpha-naphthyl acetate compared with the wild type enzyme -, 671042 3.1.1.45 C123S burst kinetics with p-nitrophenyl acetate, 10% as active as DLH 94355 3.1.1.45 C123S maximal activity 20% that of the wild type protein 94356, 94358 3.1.1.45 C123S site-directed mutagenesis, mutation of the active site cysteine to a serine, giving a catalytic triad found in serine proteases, completely changes the catalytic activity to a dienelactone isomerase -, 650835 3.1.1.45 C123S/R206A 65% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 C132A mutant shows a drastic reduction of the olmesartan medoxomil-hydrolyzing activity 704653 3.1.1.45 C132A mutant shows a low activity, 30% of wild-type 704653 3.1.1.45 C151S site-directed mutagenesis, almost inactive mutant 714312 3.1.1.45 C60S no reduction in activity 94355, 94364 3.1.1.45 D107A mutation of a residue in the putative metal-binding site of trans-DLH results in a drastic decrease in activity 702072 3.1.1.45 D107A/H111A mutation of a residue in the putative metal-binding site of trans-DLH results in a drastic decrease in activity 702072 3.1.1.45 D171N no detectable activity 94356 3.1.1.45 D198N site-directed mutagenesis, almost inactive mutant 714312 3.1.1.45 E294A mutation of a residue in the putative metal-binding site of trans-DLH results in a drastic decrease in activity 702072 3.1.1.45 E36A no detectable activity 94356 3.1.1.45 E36A/C123S no activity activity towards alpha-naphthyl acetate -, 671042 3.1.1.45 E36D 104% activity towards alpha-naphthyl acetate compared with the wild type enzyme -, 671042 3.1.1.45 E36D/C123S 183% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/C123S/A134S/S208G/A229V/K234R 256% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/C123S/A134T/A229V 164% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/C123S/A205D 217% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/C123S/A205D/A229V 222% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/C123S/A205D/A229V 224% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/C123S/A205D/A229V 230% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/C123S/A229V 187% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/C123S/F173A no activity activity towards alpha-naphthyl acetate 671042 3.1.1.45 E36D/C123S/G211D/A229V 172% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/C123S/G211D/K234N 205% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/C123S/R206T/A229V 122% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/R105H/C123S/G211D/K234N 233% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/R45Q/C123S 172% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/R45Q/C123S/A205D/A229V 148% activity towards alpha-naphthyl acetate compared with the wild type enzyme 671042 3.1.1.45 E36D/R81A/C123S/R206A 114% activity towards alpha-naphthyl acetate compared with the wild type enzyme -, 671042 3.1.1.45 E36N/C123S 140% activity towards alpha-naphthyl acetate compared with the wild type enzyme -, 671042 3.1.1.45 H111A mutation of a residue in the putative metal-binding site of trans-DLH results in a drastic decrease in activity 702072 3.1.1.45 H229N site-directed mutagenesis, almost inactive mutant -, 714312 3.1.1.45 H281A mutation of a residue in the putative metal-binding site of trans-DLH results in a drastic decrease in activity 702072 3.1.1.45 additional information the surface mutations Q110L, Y137C and N154D do not influence the kinetic properties. Mutations Q110L and N154D have stabilizing effects, the Y137C mutation alone is destabilizing. The three mutations together increase the melting temperature by 3.4 degrees -, 752118 3.1.1.45 additional information to evaluate the metal dependence of trans-DLH, a strep-tagged protein is purified 690759 3.1.1.45 Q105A mutation of a residue in the putative metal-binding site of trans-DLH results in a drastic decrease in activity 702072 3.1.1.45 Q35H/F38L/C123S/Y137C/Y145C/E199G/S208G/G211D mutations do not influence the kinetic properties -, 752118 3.1.1.45 Q35H/F38L/C123S/Y145C/E199G/S208G/G211D mutations do not influence the kinetic properties -, 752118 3.1.1.45 Q35H/F38L/C123S/Y145C/N154D/E199G/S208G/G211D mutations do not influence the kinetic properties -, 752118 3.1.1.45 Q35H/F38L/Q110L/C123S/Y137C/Y145C/N154D/E199G/S208G/G211D mutations do not influence the kinetic properties 752118 3.1.1.45 Q35H/F38L/Q110L/C123S/Y145C/E199G/S208G/G211D mutations do not influence the kinetic properties -, 752118 3.1.1.45 Q35H/F38L/Q110L/C123S/Y145C/N154D/E199G/S208G/G211D mutations do not influence the kinetic properties 752118 3.1.1.45 S102A site-directed mutagenesis, the mutant is 3-5fold less active than the wild-type enzyme -, 714312 3.1.1.45 S103A site-directed mutagenesis, 20% reduced activity compared to the wild-type enzyme -, 714312 3.1.1.45 S120A site-directed mutagenesis, the mutant is 3-5fold less active than the wild-type enzyme -, 714312 3.1.1.45 S139A site-directed mutagenesis, the mutant is 3-5fold less active than the wild-type enzyme 714312 3.1.1.45 S147A site-directed mutagenesis, 20% reduced activity compared to the wild-type enzyme 714312 3.1.1.45 S210A site-directed mutagenesis, the mutant shows similar activity like the wild-type enzyme -, 714312 3.1.1.45 Y155C site-directed mutagenesis, the enzyme mutant is caused by a single nucleotide polymorphism rs35489000 and shows 50% reduced olmesartan medoxomil-hydrolase activity, slightly higher Km, and lower Vmax compared to the wild-type protein 729629