Table 1.
Strains and plasmids used in this study
| Strain or plasmid | Characteristics | Source |
|---|---|---|
| Strains | ||
| L. lactis | ||
| TIL46 | WT strain derived from L. lactis NCDO763 cured of its 2-kb plasmid | National Collection of Food Bacteria (Shinfield, Reading, United Kingdom) |
| TIL46noxE | TIL46 noxE-negative mutant obtained by SCOa integration of pORInoxE; Eryr | 34 |
| TIL46noxE-pJIM::noxE | TIL46 noxE-negative mutant complemented with pJIM::noxE; Eryr Cmr | 34 |
| NZ9000 | MG1363; pepN::nisRK | 19a |
| NZ9000noxE | NZ9000 noxE-negative mutant obtained by SCO integration of pORInoxE; Eryr | This study |
| E. coli | ||
| TG1 repA+ | TG1 derivative with the repA gene integrated into the chromosome, allowing replication of L. lactis plasmids | P. Renault (INRA, Génétique Microbienne, Jouy-en-Josas, France) |
| Rosetta | F−ompT hsdSB (rB− mB−) gal dcm pRARE; Cmr | Novagen |
| Plasmids | ||
| pET28a | 5.4-kb, expression vector for C-terminal 6-His-tag fusion; Kanr | Novagen |
| pET28a::noxE_TIL46 | pET28a with noxE gene from TIL46 + C-terminal 6-His tag under the control of the T7 promoter | This study |
| pET28a::noxE_A144T | pET28a::noxE_TIL46 containing the A144T mutation in noxE | This study |
| pET28a::noxE_A303T | pET28a::noxE_TIL46 containing the A303T mutation in noxE | This study |
| pET28a::noxE_K384N | pET28a::noxE_TIL46 containing the K384N mutation in noxE | This study |
| pET28a::noxE_A303G | pET28a::noxE_TIL46 containing theA303G mutation in noxE | This study |
| pET28a::noxE_N302S | pET28a::noxE_TIL46 containing the N302S mutation in noxE | This study |
| pET28a::noxE_L299T | pET28a::noxE_TIL46 containing the L299T mutation in noxE | This study |
| pET28a::noxE_A300T | pET28a::noxE_TIL46 containing the A300T mutation in noxE | This study |
| pET28a::noxE_G307A | pET28a::noxE_TIL46 containing the G307A mutation in noxE | This study |
| pET28a::noxE_G307S | pET28a::noxE_TIL46 containing the G307S mutation in noxE | This study |
| pET28a::noxE_CSK1382 | pET28a with noxE gene from CSK1382 + C-terminal 6-His tag under the control of the T7 promoter | This study |
| pET28a::noxE_T303A | pET28a::noxE_CSK1382 containing the T303A mutation in noxE | This study |
| pNZ8048 | nisAp + NcoI + MCS + terminator | 19a |
| pNZ8048::noxE_TIL46 | pNZ8048 with noxE gene of TIL46 under nisAp control | This study |
| pNZ8048::noxE_CSK1382 | pNZ8048 with noxE gene of CSK1382 under nisAp control | This study |
| pNZ8048::noxE_6His | pNZ8048 with noxE gene of TIL46 + C-terminal 6-His tag under nisAp control | This study |
| pNZ8048::noxE_A303T_6His | pNZ8048::noxE_6His containing the A303T mutation in noxE | This study |
a
SCO, single-crossover.