TABLE 2.

Strains and plasmids utilized in this study

E. coli strain or plasmidGenotypeUtilitySource or reference
Strains
    DH5αFΦ80 lacZΔM15 Δ(lacZYA-argF) U169 hsdR17 (rk, mk+) recA1 endA1 deoR thi-1 supE44 gyrA96 relA1 λEnzyme productionTOYOBO
    XL1-Red endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac mutD5 mutS mutT Tn10 (Tetr)In vivo mutagenesisTOYOBO
    MV1184 ara Δ(lac-proAB) rpsL thi (lacZΔM15) Δ(srl-recA) 306::Tn10 (Tetr)/F′ [traD36 proAB+lacIqlacZΔM15]Site-directed mutagenesisTakara Shuzo
Plasmids
    pUTE500k′Expression vector; Apr Kmr12
    pFA5FAOX-C gene coding; Apr15
    pFA101Expression of FAOX-C gene and template of mutagenesis; Apr KmrThis work
    pFAH11Positive clone of 1st round of directed evolution; Apr KmrThis work
    pFAH21Positive clone of 2nd round of directed evolution; Apr KmrThis work
    pFAH22Positive clone of 2nd round of directed evolution; Apr KmrThis work
    pFAH31Positive clone of 3rd round of directed evolution; Apr KmrThis work
    pFAH41Positive clone of 4th round of directed evolution; Apr KmrThis work
    pBluescriptII SK+For sequence of mutant FAOX genes; AprTOYOBO
    pKF19kFor site-directed mutagenesis; KmrTakara Shuzo
    pFA19kFAOX-C gene in pKF19k for site-directed mutagenesis; KmrThis work
    pFAT60AkHarboring T60A mutation derived from pFA19k; KmrThis work