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Table I. List of all P450 BM3 variants generated by site-directed mutagenesis and random mutagenesis

Name of mutant Position of amino acid substitution Amino acid change Codon change Method for diversity generation
P450 BM3 F87A 87 F->A TTT->GCC Site-directed mutagenesis
P450 BMP F87A 87 F->A TTT->GCC Site-directed mutagenesis
P450 BM3 M1 47, 87 R->F, F->A CGT->TTT, TTT->GCC Site-directed saturation mutagenesis
P450 BM3 M2 47, 87, 354 R->F, F->A, M->S CGT->TTT, TTT->GCC, ATG->TCT Site-directed saturation mutagenesis
P450 BMP M2 47, 87, 354 R->F, F->A, M->S CGT->TTT, TTT->GCC, ATG->TCT Site-directed mutagenesis
P450 BM3 M3 47, 87, 354, 363 R->F, F->A, M->S, D->H CGT->TTT, TTT->GCC, ATG->TCT, GAC->CAC Site-directed mutagenesis
P450 BM3 M4 47, 87, 354, 281 R->F, F->A, M->S, V->G CGT->TTT, TTT->GCC, ATG->TCT, GTG->GGG Site-directed mutagenesis
P450 BM3 M5 47, 87, 281, 354, 363 R->F, F->A, V->G, M->S, D->H CGT->TTT, TTT->GCC, GTG->GGG, ATG->TCT, GAC->CAC Random mutagenesis (two rounds of epPCR)
P450 BMP M5 47, 87, 281, 354, 363 R->F, F->A, V->G, M->S, D->H CGT->TTT, TTT->GCC, GTG->GGG, ATG->TCT, GAC->CAC Site-directed mutagenesis

Given are the chosen names for the mutants used in the manuscript, positions of amino acids substitutions, mutations in the corresponding codons, and the method used for diversity generation.





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