MOESM1 of Furfural tolerance and detoxification mechanism in Candida tropicalis

Additional file 1: Table S1. Strains and plasmids used in this study. Table S2. Primers used in this study. Figure S1. Furfural tolerance test evaluated by methylene blue staining. Furfural of 1, 3, 5, 7, and 9 g/L was added into the culture at mid-exponential phase. After having been stained by Methylene blue solvent 30 min, cells were observed and photoed by optical microscope. Figure S2. PCR confirmation of the specific integration in sequential ctADH1 disruption. Lane M, DNA makers; Lane A, PCR from C. tropicalis YE genome with primers ADH1-F and ADH1-R resulting in the band of 1.2 kb (ADH1); Lane B, PCR from C. tropicalis Y1 genome with primers ADH1-F and ADH1-R resulting in the band of 1.2 and 4.3 kb (ADH1 and ADH1a-HisG-URA3-HisG-ADH1b); Lane C, PCR from C. tropicalis Y2 genome with primers ADH1-F and ADH1-R resulting in the band of 1.2 and 1.7 kb (ADH1 and ADH1a-HisG-ADH1b); Lane D, PCR from C. tropicalis Y4 genome with primers ADH1-F and ADH1-R resulting in the band of 1.7 and 2.9 kb (ADH1a-HisG-ADH1b and ADH1-URA3). Figure S3. Sensitivity experiment of C. tropicalis T4, T3, T2, and YE (parent strain). Cells of C. tropicalis T4, T3, T2, and YE were inoculated into 5 ml YPD medium containing 3 g/L furfural. The cultures were incubated for 10 h at 30 °C. Figure S4. SDS-PAGE of alcohol dehydrogenase 1 expressed in E. coli. Lane M, protein molecular weight markers (Thermo Scientific, #26610, USA); Lane A, E. coli PCA cells after IPTG induction; Lane B, E. coli PCA cells before IPTG induction; Lane C, E. coli PC cells without IPTG induction; Lane D, E. coli PEA cells after IPTG induction; Lane E, E. coli PEA cells before IPTG induction; Lane F, E. coli PE cells without IPTG induction. Molecular weight of alcohol dehydrogenase 1 is around 43 kDa. Figure S5. In vivo furfural degradation in recombined E. coli. (A) Furfural degradation of E. coli PCA (with pCS-ADH1) and PC (with pCS-27) in M9 medium; (B) Furfural degradation of E. coli PEA (with pET-ADH1) and PE (with pETDuet-1) in M9 medium.