TABLE 1.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant characteristic(s)Source or reference
E. coli strains
    BL21(DE3)F ompT gal dcm lon hsdSB(rB mB) λ(DE3)20
    DH1endA1 recA1 gyrA96 thi-1 glnV44 relA1 hsdR17(rK mK+) λ13
    LT-ΔfadEDH1 ΔfadE with pKS118
    EGS084LT-ΔfadE with pEC-XK99EThis study
    EGS145LT-ΔfadE with pEG142This study
    EGS180LT-ΔfadE with pEG174This study
    EGS210LT-ΔfadE with pEG200This study
    EGS212LT-ΔfadE with pEG205This study
    EGS220BL21(DE3) with pEG185This study
    EGS244LT-ΔfadE with pEG225This study
    EGS300LT-ΔfadE with pEG275This study
Micrococcus luteus ATCC 4698Wild typeATCC
Plasmids
    pEC-XK99EKmr; E. coli-C. glutamicum shuttle expression vector with ColE1 origin of replication and trc promoter11
    pKS1Cmr; p15a derivative containing ‘tesAa under the lacUV5 promoter18
    pSKB3Kmr; derivative of expression vector pET-28a with the thrombin protease site replaced by a TEV protease siteStephen K. Burley
    pEG142Kmr; ∼5.2 kb containing Mlut_13230-13250 cloned into pEC-XK99E at KpnI and XbaI sitesThis study
    pEG174Kmr; ∼1 kb containing Mlut_13230 #1 into pEC-XK99E at EcoRI and XbaI sitesThis study
    pEG185Kmr; ∼1-kb fragment of Mlut_13230 #2 cloned into pSKB3 at NdeI and SalI sitesThis study
    pEG200Kmr; ∼1.2-kb fragment of Mlut_09290 cloned into pEC-XK99E at EcoRI and XbaI sitesThis study
    pEG205Kmr; ∼1-kb fragment of Mlut_09310 cloned into pEC-XK99E at EcoRI and XbaI sitesThis study
    pEG225Kmr; ∼3-kb fragment of Mlut_13240 cloned into pEC-XK99E at XbaI siteThis study
    pEG275Kmr; ∼1.1-kb fragment of Mlut_13250 cloned into pEC-XK99E at XbaI and SbfI sitesThis study
  • a ‘tesA, tesA thoiesterase gene from E. coli from which the region encoding the leader sequence has been removed.