Supporting Information Metabolomics reveals stage
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1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 Supporting Information Metabolomics reveals stage-specific metabolic pathways of microbial communities in two-stage anaerobic fermentation of corn-stalk Dawei Yang1,2, Xiaolei Fan1*, Xiaoshuang Shi1, Shujuan Lian1, Jiangtao Qiao1, Rongbo Guo1* 1. Key laboratory of Biofuels, Shandong Industrial Engineering Laboratory of Biogas Production & Utilization, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, 189 Songling Road, Qingdao 266101, P.R. China 2. Central Laboratory, Liaocheng People's Hospital, Liaocheng, Shandong, 252000, P. R. China *Corresponding author: Xiaolei Fan, Ph. D; Rongbo Guo, Ph. D Key Laboratory of Biofuels,Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences No.189 Songling Road, Qingdao, Shandong Province 266101, P. R. China Phone: +86-532-80662708 Fax: +86-532-80662708 Email address: guorb@qibebt.ac.cn fanxl@qibebt.ac.cn 1 1 Additional method: Control samples 2 Ten ml of fermentation broth without quenching was centrifuged at 10000 x g for 5 min at 4 °C. The 3 supernatant was rapidly filtered through a 0.22 µm to remove microbial cells as control sample to 4 determine the level of contamination from fermentation broth. 5 6 2 1 2 Supplementary Table 1 List of identified metabolites in the two-stage fermentation process. Compounds Pubchem CIDa KEGGb Valeric acid Urea Uracil Trehalose Ferulic acid Succinic acid Stearic acid Pyroglutamic acid Pyrocatechol Putrescine Acrylic acid Pipecolic acid Phytol Phytanic acid 4-Hydroxyphenylacetate Phosphoric acid Phenylacetic acid Pelargonic acid Methyl palmitate Palmitic acid Oxalic acid Oleic acid Myristic acid Monostearin Methylsuccinic acid Melibiose Coumaric acid Mannitol Malic acid L-Valine L-Tyrosine L-Threonine L-Serine L-Proline L-Methionine L-Lysine L-Leucine L-Lactic acid L-Isoleucine L-Glutamine L-Alpha-aminobutyric acid Palatinitol Isocaproic acid Indoleacetic acid Hydroxylamine Hydrocinnamic acid 7991 1176 1174 1143 445858 1110 5281 7405 289 1045 6581 849 5280435 26840 127 1004 999 8158 8181 985 971 965 11005 24699 10349 440658 637541 6251 525 1182 6057 6288 5951 145742 6137 5962 6106 107689 791 5961 80283 3034828 12587 802 787 107 C00803 C00086 C00106 C01083 NA C00042 C01530 C01879 C00090 C00134 C00511 C00408 C01389 C01607 C00642 C00013 C07086 C01601 C16995 C00249 C00209 C00712 C06424 NA C08645 C05402 C12621 C00392 C00149 C00183 C00082 C00188 C00065 C00148 C00073 C00047 C00123 C00186 C00407 C00064 C02356 NA C03467 C00954 C00192 C05629 3 Hexanoylcarnitine Heptanoic acid Heptadecanoic acid 9-Heptadecenoic acid Glycine Glycerol-3-phosphate Glycerol Glyceric acid Glutaric acid Gluconic acid Fumaric acid Ethylene glycol Ethanolamine Dodecanoic acid Dimethylglycine D-Glucose D-Galactose Aspartic acid D-Arabitol Citrulline Decanoic acid Caprylic acid Cadaverine Butyric acid Butylamine Benzoic acid Azelaic acid Arachidic acid Alanine Adenosine Adenine Acetic acid 5-Hydroxyhexanoic acid 5-Aminopentanoic acid 4-Hydroxybutyric acid 3-Hydroxyvaleric acid 3-Hydroxyphenylacetic acid 2-Phenylacetamide 2,4-dihydroxybenzoic acid 10-Nonadecenoic acid 6426853 8094 10465 5282748 750 439162 753 439194 743 10690 723 174 700 3893 673 5793 439357 83887 94154 9750 4225619 379 273 264 8007 243 2266 10467 5950 6083 190 176 170748 138 10413 107802 12122 7680 1491 5312513 NA C05799 NA NA C00037 C00093 C00116 C00258 C00489 C00257 C00122 C01380 C00189 C02679 C01026 C00031 C00124 C00402 C01904 C00327 C02530 C06423 C01672 C00246 C18706 C00180 C08261 C06425 C01401 C00212 C00147 C00033 NA C00431 C00989 NA C05593 C02505 C00230 NA 1 a CID: compound identification number from PubChem 2 b compound number from Kyoto Encyclopedia of Genes and Genomes (KEGG) 4 1 Figure Legends: 2 Supplementary Fig. 1 A number of representative metabolites from different chemical classes was 3 compared of the different extraction methods based on ANOVA analysis. Data distributions were 4 displayed by box-whisker plats, giving the arithmetic mean value for each category, the standard error 5 as box, and whiskers for 1.96 times the category standard error to indicate the 95 % confidence intervals, 6 assuming normal distributions. 7 8 5 6
