Additional file 1 of Expressing 2-keto acid pathway enzymes significantly increases photosynthetic isobutanol production

Additional file 1. Fig. S1: Schematic overview of genetic constructs used and Western-immunoblot results of engineered Synechocystis PCC 6803 strains HX0 and HX6. (A) Schematic presentation of the genetic constructs in the engineered strains. kivdS286T: encodes α-ketoisovalerate decarboxylase (Lactococcus lactis). alsS: encodes acetolactate synthase (Bacillus subtilis). KivdS286T expressed on self-replicating vectors was Strep-tagged at the N-terminal; AlsS expressed in the ddh (slr1556) site of chromosome was His-tagged at the N-terminal. (B) Western-immunoblot results of strains HX0 and HX6. Each lane represents result from respective strain. 5 μg and 20 μg of total soluble protein were loaded for each strain to detect Strep-tagged KivdS286T and His-tagged AlsS, respectively. Fig. S2: Comparison of growth in engineered Synechocystis PCC 6803 strains HX0, HX5, HX7, HX8, and HX9 during 8-day cultivation. Results are the mean of three biological replicates, each with three technical replicates. Error bars represent standard deviation. Fig. S3: Schematic overview of genetic constructs used and comparison of molar ratio of isobutanol and 3-methyl-1-butanol (3M1B) of engineered Synechocystis PCC 6803 strains HX15, HX29, and HX45. (A) Molar ratio of isobutanol and 3M1B of indicated strains, calculated based on the isobutanol production measured on day 4. (B) Schematic presentation of the genetic constructs in the engineered strains. Asterisk represents significant difference between HX45 and HX15 (One-way ANOVA, p < 0.05). Results are the mean of three biological replicates, each with three technical replicates. Error bars represent standard deviation. Table S1: Sequences of codon optimized synthetic genes used in this study. Table S2: Plasmids used in this study. Expressed genes in bold. Table S3: Oligonucleotides used in this study. Table S4: Expression quantification of heterologously expressed enzymes. The expression level of each protein is presented by the corresponding band intensity. The unit is intensity x mm