Figure 1.
Overview of the pathway introduced to S. cerevisiae for 2-butanol production.
The heterologous proteins expressed are marked in red. In violet is the Ado-Cbl recycling system. X-Cbl is an unidentified complex of cobalamine made due to inactivation [34].
Table 1.
List of strains and DNA constructs used in this study.
Figure 2.
Constructs required for B12 dependent 2-butanol production in yeast.
Depiction of the 4 constructs made for pduCDE, pduGH and SADH respectively. The pduCDE construct was made with and without the SacI fragment containing the TEV-protease.
Figure 3.
Verification of the TEV-strategy by western analysis.
Western blot analysis of the control strain where no heterologous protein of 2-butanol pathway was expressed (strain YSH6.142-3D - Column A) and the strain with the entire set for TEV expression system (pduCDE+pduGH) together with the secondary alcohol dehydrogenase (Sadh) (strain yPG-3 - Column B). Data is shown for W303 Δgpd1,2 background.
Figure 4.
Comparison of HPLC results for both the control and constructed strain.
The graph is zoomed in to the time span where 2-butanol and 2-butanone are known to appear. The base-line used to calculate the amount of butanone and butanol is indicated as a dashed line.