Table 2 Plasmids and strains constructed in this study
From: Metabolic engineering of methylotrophic Pichia pastoris for the production of β-alanine
Names | Relative characteristics | References |
|---|---|---|
Plasmids | ||
pPICZA | Vector for extracellular expression recombinant protein carrying ZeoR | Invitrogen |
pAOα | Vector for extracellular expression; derived from pPICZαA and pAO815, HIS4 + | Zhu et al. (2011) |
pMPICZHis | Extracellular expression vector carrying HIS4 gene, ZeoR | This work |
pMPICKmHis | Replace antibiotic resistance marker (Zeocin) of pMPICZHis with kanamycin | This work |
pMPIC2H | Replace antibiotic resistance marker (Zeocin) of pMPICZHis with hygromycin | This work |
pMPICZHis-bliADC | Codon-optimized panD gene from Bacillus paralicheniformis ATCC 9945a cloned into pMPICZHis at BstBI and NotI site | This work |
pMPICZHis-bsADC | Codon-optimized panD gene from Bacillus subtilis cloned into pMPICZHis at BstBI and NotI site | This work |
pMPICZHis-sgeADC | Codon-optimized panD gene from Streptomyces griseorubiginosus cloned into pMPICZHis at BstBI and NotI site | This work |
pMPICZHis-cguADC | Codon-optimized panD gene from Corynebacterium glutamicum cloned into pMPICZHis at BstBI and NotI site | This work |
pMPICZHis-TcCSADC | Cysteine sulfinic acid decarboxylase from Tribolium castaneum (TcCSADC) cloned into pMPICZHis at BstBI and NotI site | This work |
pMPICZHis-Oan | aspDH gene from Ochrobactrum anthropi ATCC 4918 cloned into pMPICZHis at BstBI and NotI site | This work |
pMPICZHis-Spe | aspDH gene from Serratia proteamaculans cloned into pMPICZHis at BstBI and NotI site | This work |
pMPICZHis-ADC-Oan | panD expression cassette (pMPICZHis-ADC digest by SphI and BamHI) cloned into pMPICZHis-Oan at SphI and BglII site | This work |
pMPICZHis-ADC-Spe | panD expression cassette (pMPICZHis-ADC digest by SphI and BamHI) cloned into pMPICZHis-Spe at SphI and BglII site | This work |
pMPICKmHis-AAT1 | Cytosolic aspartate aminotransferase from P. pastoris GS115 cloned into pMPICZHis at BstBI and NotI site | This work |
pMPICKmHis-AAT2 | Mitochondrial aspartate aminotransferase from P. pastoris GS115 cloned into pMPICZHis at BstBI and NotI site | This work |
pMPICZHis-2ADC | Two copies of panD gene expression cassette cloned into pMPICZHis | This work |
pMPICZHis-2ADC-Spe | Two copies of panD gene expression cassette and one copy of SpeaspDH gene expression cassette cloned into pMPICZHis | This work |
pMPIC2H-PPC | ppc gene from Escherichia coli cloned into integration vector pMPIC2H at BstBI and NotI site | This work |
pMPIC2H-AsPCK | pck gene from Actinobacillus succinogenes cloned into integration vector pMPIC2H at BstBI and NotI site | This work |
pMPIC2H-PCK | pck gene from Escherichia coli cloned into integration vector pMPIC2H at BstBI and NotI site | This work |
Strains | ||
E. coli DH5α | Commercial transformation host for cloning | Invitrogen |
P. pastoris GS115 | Commercial transformation host for cloning; HIS4-, Mut + | Invitrogen |
Δku70 | GS115, ΔKU70; HIS4 + | This work |
BliADC | Δku70 harboring pMPICZHis-bliADC | This work |
BsADC | Δku70 harboring pMPICZHis-bsADC | This work |
SgeADC | Δku70 harboring pMPICZHis-sgeADC | This work |
CguADC | Δku70 harboring pMPICZHis-cguADC | This work |
TcCSADC | Δku70 harboring pMPICZHis-TcCSADC | This work |
ADC-Oan | Δku70 harboring pMPICZHis-ADC-Oan | This work |
ADC-Spe | Δku70 harboring pMPICZHis-ADC-Spe | This work |
ADC-AAT1 | Δku70 harboring pMPICZHis-bsADC and pMPICKmHis-AAT1 | This work |
ADC-AAT2 | Δku70 harboring pMPICZHis-bsADC and pMPICKmHis-AAT2 | This work |
2ADC | Δku70 harboring pMPICZHis-2ADC | This work |
2ADC-Spe | Δku70 harboring pMPICZHis-2ADC-Spe | This work |
PPC | Δku70 harboring pMPICZHis-2ADC-Spe and pMPIC2H-PPC | This work |
AsPCk | Δku70 harboring pMPICZHis-2ADC-Spe and pMPIC2H-AsPCK | This work |
PCK | Δku70 harboring pMPICZHis-2ADC-Spe and pMPIC2H-PCK | This work |