Additional file 1 of Engineering microbial consortia of Elizabethkingia meningoseptica and Escherichia coli strains for the biosynthesis of vitamin K2

Additional file 1: Figure S1. Terpenoid backbone biosynthesis of E. meningoseptica sp. F2 based on KEGG pathway assignment. A-CoA acetyl coenzyme A; AA-CoA acetoacetyl-CoA; HMG-CoA hydroxymethylglutaryl-CoA; IPP isopentenyl diphosphate; DMAPP dimethylallyl diphosphate; GPP geranyl diphosphate; FPP farnesyl diphosphate; GGPP geranylgeranyl diphosphate; OPP octaprenyl diphosphate. Figure S2. Chromatography of MK-n (n = 4, 5, 6, 7, 8) by HPLC system. A The sample was extracted from monoculture system of E. meningoseptica, B the sample was extracted from coculture system of E. meningoseptica and E. coli. Figure S3. A The purification of GB1-OPPS. M: protein molecular weight marker; lane 1: The total cell lysate supernatant flows through the column. Lanes 2���6: wash fractions with 10, 15, 20, 25, 30 mM imidazole, respectively. Lanes 7���8: Elution fractions with 250 mM imidazole, respectively. B Purified GB1-EmOPPS proteins was confirmed by mass spectrometry. Figure S4. Enzymatic assay of EmOPPS in vivo and vitro. A, B LC-MS analysis of the GB1-EmOPPS products with farnesyl diphosphate (FPP) and isopentenyl diphosphate (IPP) as substrates after hydrolyzation. C, D J01 strains could accumulated MK-n (n = 6, 7, 8). LC-MS analysis of the production of this strains. A, C Horizontal graphs refer to result of HPLC and vertical graphs B, D refer to mass spectra. Figure S5. The comparison of the mass spectrum of the fermentation results of the J01, J02 strain with that of the MK-n (n = 4, 5, 6, 7) standard, A MK-4, MK-5, MK-6, and MK-7 standard, respectively, for MS analysis. B The mass spectrum results of monocultured E. coli producing MK-8 as a reference. Figure S6. Analysis and identification of mevalonolactone by GC-MS from fermentation supernatant. Figure S7. The monoculture of different E. coli strains as comparison. The error bars represent the standard error of at least three biological replicates. Statistical significance (p < 0.01) compared to the original strains. Figure S8. Visualization of different inocula by confocal laser scanning microscope. A E. meningoseptica. B E. coli. C, D E. meningoseptica and E. coli co-culture. Red circles surround the connected E. meningoseptica and E. coli cells in view. C Bright field; D fluorescence image merged image. Table S1. Masses and identities of LC/MS analyses results. Table S2. Oligonucleotide primers used in this study.