Table 1 Bacterial strains and plasmids

B. subtilis NZ8900

B. subtilis 168 strain amyE::spaRK

MoBiTec GmbH

B. subtilis NZ8963

ATCC6633 subtilin producing strain

MoBiTec GmbH

B. subtilis RIK1285

Marburg 168 derivative: trpC2, lys1, aprE Δ3, nprR2, nprE18

Takara Bio Inc.

E. coli JM109

Intermediate cloning host endA1 glnV44 thi-1 relA1 gyrA96 recA1 mcrB + Δ(lac-proAB) e14- [F’ traD36 proAB + lacIq lacZΔM15] hsdR17(rK-mK+)

Lab stock

E. coli MC1061

Intermediate cloning host F- araD139 Δara-leu)7696 Δ(lac)X74 galU galK hsdR2 mcrA mcrB1 rspL

MoBiTec GmbH

S. lividans TK24

[42]

KU Leuven

Plasmids

Genotype or characteristics

pNZ8901

Subtilin regulated expression (SURE) vector P_spaSmut, Cm^R

E. coli MC1061/

B. subtilis NZ8900

MoBiTec GmbH

pNZ-Bs SP AmDyP

pNZ containing signal peptide (SP) from B. subtilis NZ8900 dye-decolorizing peroxidase (DyP), DyP gene from Amycolatopsis 75iv2 (AmDyP), Tobacco Etch Virus cleavage site (TEV), 6 Histidine tag (6His)

E. coli MC1061 /

B. subtilis NZ8900

This study

pNZ-AmDyP

pNZ containing gene of interest AmDyP, TEV, 6His

E. coli MC1061 /

B. subtilis NZ8900

This study

pNZ-ScLac

pNZ containing gene of interest small laccase (Slac) from Streptomyces coelicolor (ScLac), TEV, 6His

E. coli MC1061 /

B. subtilis NZ8900

This study

pNZ-ScLac ∆SP

pNZ containing gene of interest ScLac with the deletion of the SP, TEV, 6His

E. coli MC1061 /

B. subtilis NZ8900

This study

pBE-S

pUB ori, Kan^R, ColE1 ori, Amp^R, aprE promoter, aprE SP, His tag

E. coli JM109 /

B. subtilis RIK1285

Takara Bio Inc.

pBE-Bs SP AmDyP

pBE containing B. subtilis SP, AmDyP, TEV, 6His

E. coli JM109 /

B. subtilis RIK1285

This study

pBE-AmDyP

pBE containing gene of interest AmDyP, TEV, 6His

E. coli JM109 /

B. subtilis RIK1285

This study

pBE-AmLac

pBE containing gene of interest Slac from Amycolatopsis 75iv2 (AmLac), TEV, 6His

E. coli JM109 /

B. subtilis RIK1285

This study

pBE-AmLac ∆SP

pBE containing gene of interest AmLac with the deletion of the SP, TEV, 6His

E. coli JM109 /

B. subtilis RIK1285

This study

pBE-ScLac

pBE containing gene of interest ScLac, TEV, 6His

E. coli JM109 /

B. subtilis RIK1285

This study

pBE- ScLac ∆SP

pBE containing gene of interest ScLac with the deletion of the SP, TEV, 6His

E. coli JM109 /

B. subtilis RIK1285

This study

pBE-SvLac

pBE containing gene of interest SLAC from Streptomyces viridosporus (SvLac), TEV, 6His

E. coli JM109 /

B. subtilis RIK1285

This study

pBE- SvLac ∆SP

pBE containing gene of interest SvLac with the deletion of the SP, TEV, 6His

E. coli JM109 /

B. subtilis RIK1285

This study

pBE- ScLac ∆SP-TEV-AmDyP

pBE containing ScLac with the deletion of the SP, TEV, AmDyP, TEV, 6His

E. coli JM109 /

B. subtilis RIK1285

This study

pBE- ScLac ∆SP-TEV-GFP

pBE containing ScLac with the deletion of the SP, TEV, GFP, TEV, 6His

E. coli JM109 /

B. subtilis RIK1285

This study

pIJ486

P_vsi, Rep ori, Ts^R, Kan^R

S. lividans TK24

KU Leuven

pIJ- ScLac

pIJ containing ScLac, TEV, 6His

S. lividans TK24

This study

pIJ- ScLac ∆SP

pIJ containing ScLac with the deletion of the SP, TEV, 6His

S. lividans TK24

This study

pIJ- AmLac

pIJ containing AmLac, TEV, 6His

S. lividans TK24

This study

pIJ- SvLac

pIJ containing SvLac, TEV, 6HIs

S. lividans TK24

This study

pIJ-Sc SP-AmDyP

pIJ containing SP from ScLac, AmDyP, TEV, 6His

S. lividans TK24

This study

pUC19-P_vsi

P_vsi, Rep ori, Amp^R, Amp^R promoter, CAP binding site, lac promoter, lacZα

E. coli JM109

[39]

pUC-ScLac

pUC containing P_vsi, ScLac, TEV, 6His

E. coli JM109

This study

pUC-ScLac ∆SP

pUC containing P_vsi, ScLac with the deletion of the SP, TEV, 6His

E. coli JM109

This study

pUC-AmLac

pUC containing P_vsi, AmLac, TEV, 6His

E. coli JM109

This study

pUC-SvLac

pUC containing P_vsi, SvLac, TEV, 6His

E. coli JM109

This study

pUC-Sc SP-AmDyP

pUC containing P_vsi, SP from ScLac, AmDyP, TEV, 6His

E. coli JM109

This study