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Fig. 5 | Microbial Cell Factories

Fig. 5

From: Restoration of biofuel production levels and increased tolerance under ionic liquid stress is enabled by a mutation in the essential Escherichia coli gene cydC

Fig. 5

Production of the Biofuel Precursor Limonene and Isopentenol Under Exogenous [EMIM]OAc Growth Conditions. a E. coli DH1 cells with or without the cydC-D86G mutation at the genomic locus and a limonene production plasmid (JBEI-6409) were tested for limonene production in the presence or absence of 100 mM [EMIM]OAc (“Methods”). Time points were sampled 24 h or 48 h after pathway induction with 15 µM IPTG. Black bars; WT strains. Grey bars; cydC-D86G strains. Orange bars; ∆rcdA strains. b E. coli DH1 ∆ptapoxBackA (JBEI-3606) harboring [pTE50/PcydD-cydD-cydC] (white bars), [pTE88/PcydD-cydD-cydC-D86G] (red striped bars), or [pTE100 PcydD-cydD-cydC-D86V] (blue bars) along with an isopentenol production plasmid (JBEI-9321) were prepared for biofuel production in the presence of 75 mM [EMIM]OAc. Sampling was conducted at 24 h or 48 h after pathway induction with 500 µM IPTG. Production of isopentenol was compared to E. coli DH1 strains harboring an empty plasmid (PcydD only) and the isopentenol production plasmid (JBEI-9321) in the absence of [EMIM]OAc. Only trace isopentenol production (with high acetate formation) was observed in the presence of [EMIM]OAc using either E. coli DH1 or E. coli DH1 cydC-D86G strains (data not shown)

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