MOESM1 of Metabolic engineering of Clostridium beijerinckii to improve glycerol metabolism and furfural tolerance

Additional file 1: Table S1. Similarities (%) between Cp and Cb Gldh protein sequences. NCBI Blastp algorithm was used for alignment. Table S2. Root mean square deviation (RMSD) values for secondary structure alignment of Cp Gldhs. Table S3. Comparison of DhaK protein sequences between Cp and Cb DhaK proteins. NCBI Blastp algorithm was used for alignment. Table S4. List of primers and PCR protocols used to generate constructs. Figure S1. Schematics of some recombinant plasmids that were used to transform electrocompetent Cb. Figure S2. Enzyme–substrate conduit demonstrating putative metabolic function of Cp glycerol catabolic pathway engineering in Cb. Engineering of two Gldh as fused protein was designed to increase the active site concentration per location, thereby improving the efficiency of NAD(P)H generation during glycerol catabolism. DHA dihydroxyacetone, DHAP DHA phosphate, DhaK DHA kinase, DhaD1 and GldA1 Cp Gldh. Figure S3. DNA electrophoresis gel image to confirm PCR amplicons. Lanes (1) dhaD1, (2) gldA1, (3) dhaD1+gldA1, (4) DNA ladder, (5) dhaK, and (6) [dhaD1+gldA1] + dhaK. Figure S4. Furfural detoxification profile of C. berijerinckii-pWUR460_dhaD1+gldA1 during the fermentation of glucose + glycerol (1: 2 molar ratio) challenged with 3 g/L (A), 4 g/L (B), 5 g/L (C), and 6 g/L furfural (D). (FF: Furfural; FA: Furfuryl alcohol).