Figure 4From: Cytosolic re-localization and optimization of valine synthesis and catabolism enables increased isobutanol production with the yeast Saccharomyces cerevisiaeComplementation experiments of triple ilv deletion mutants. Isoy16 was transformed with plasmids expressing genes for wild-type and truncated Ilv2ΔN54, Ilv5Δ48 and Ilv3Δ19 enzymes, respective combinations, and codon-optimized genes for truncated Ilv2ΔN54, Ilv5Δ48 and Ilv3Δ19 proteins. Transformants were grown on selective SMD + leu-val + ile medium, washed with sterile water and spotted in serial dilutions on selective SMD medium agar plates containing or lacking valine or isoleucine, respectively, and incubated for up to 7 days at 30°C (detailed description in Methods). Cells containing empty vectors were pregrown on selective SMD + leu + val + ile medium. Panel A: Isoy16 containing empty vectors or plasmids encoding for wild-type or truncated Ilv2, Ilv5 and Ilv3, respective combinations. Panel B: Isoy16 expressing plasmid p425-synthILV235. Isoy16 transformed with empty vector were spotted as a negative control.Back to article page