Table 2.
Strains and plasmids used in this study
| Strain | Description | Reference | |
|---|---|---|---|
| Escherichia coli DH5α | General cloning host, F-thi-1 endA1 hsdR17(r-, m-) supE44 _lacU169 (_80lacZ_M15) recA1 gyrA96 relA1 | Stratagene/Genomics Agilent | |
| Bacillus methanolicus MGA3 | Wild type strain | ATCC53907 | |
| B. licheniformis MW3 | Bacillus licheniformis DSM13 (ΔhsdR1, ΔhsdR2) | [100] | |
| B. subtilis 168 | Wild type strain | ATCC23857 | |
| Geobacillus stearothermophilus 10 | Wild type strain | DSM13240 | |
| Plasmid | Strain abbreviationa | Description | Reference |
|---|---|---|---|
| pBV2xp | EV | KmR and ApR; pHCMC04 derivative, gene expression under the control of the inducible xylose promoter from B. megaterium, theta replicating | [83] |
| Protease gene carrying strains/plasmids | |||
| pBV2xp-aprBl | spPBl-aprBl | KmR and ApR; pBV2xp derivative for expression of apr from B. licheniformis with native signal peptide (locus tag BLI_RS05490) | This study |
| pBV2xp-aprBs | spPBs-aprBs | KmR and ApR; pBV2xp derivative for expression of aprE from B. subtilis with native signal peptide (locus tag GP2222_11200) | This study |
| pBV2xp-aprGs | spPGs-aprGs | KmR and ApR; pBV2xp derivative for expression of aprGs from G. stearothermophilus with native signal peptide (locus tag GT50_00190) | This study |
| α-amylase gene carrying strains/plasmids | |||
| pBV2xp-spBl-amyBl | spBl-amyBl | KmR and ApR; pBV2xp derivative for expression of amyL from B. licheniformis with native signal peptide (locus tag BL00499) | This study |
| pBV2xp-spBs-amyBs | spBs-amyBs | KmR and ApR; pBV2xp derivative for expression of amyE from B. subtilis with native signal peptide (locus tag BSU03040) | This study |
| pBVxp-spGs-amyGs | spGs-amyGs | KmR and ApR; pBV2xp derivative for expression of amyS from G. stearothermophilus with native signal peptide (locus tag GT50_07800) | This study |
| pBV2xp-spBm-amyBm | spBm-amyBm | KmR and ApR; pBV2xp derivative for expression of amy from B. methanolicus with native signal peptide (locus tag BMMGA3_04345) | This study |
| pBV2xp-spGs-amyBs | spGs-amyBs | KmR and ApR; pBV2xp derivative for expression of amyE from B. subtilis with α-amylase signal peptide from G. stearothermophilus | This study |
| pBVxp-spGs-amyBl | spGs-amyBl | KmR and ApR; pBV2xp derivative for expression of amyL from B. licheniformis with α-amylase signal peptide from G. stearothermophilus | This study |
| sfGFP gene carrying strains/plasmids | |||
| sfGFP-pBAD | AmR; pBAD/His derivative for expression of sfGFP under the control of ParaBAD | sfGFP-pBAD was a gift from Michael Davidson and Geoffrey Waldo (Addgene plasmid # 54519) [82] | |
| pBV2xp-sfGFP | sp0-sfGFP | KmR and ApR; pBV2xp derivative for expression of sfGFP from sfGFP-pBAD under control of the xp promoter | This study |
| pBV2xp-spGs-sfGFP | spGs-sfGFP | KmR and ApR; pBV2xp-sfGFP derivative with amylase signal peptide from G. stearothermophilus | This study |
| pBV2xp-spBl-sfGFP | spBl-sfGFP | KmR and ApR; pBV2xp-sfGFP derivative with amylase signal peptide from B. licheniformis | This study |
| pBV2xp-spBs-sfGFP | spBs-sfGFP | KmR and ApR; pBV2xp-sfGFP derivative with amylase signal peptide from B. subtilis | This study |
| pBV2xp-spBm-sfGFP | spBm-sfGFP | KmR and ApR; pBV2xp-sfGFP derivative with amylase signal peptide from B. methanolicus | This study |
aAbbreviation used for B. methanolicus strain MGA3 carrying the listed plasmid. spP: signal peptide preceding protease; sp: signal peptide preceding α-amylase. Example: Strain spBl-amyBl carries plasmid with (α-amylase) signal peptide from B. licheniformis, and α-amylase gene from B. licheniformis
KmR kanamycin resistance marker, ApR ampicillin resistance marker