Supplementary figure legends Fig. S1. Lineweaver-Burk plot of putrescine uptake by YeeF. An overnight culture of SK629 was inoculated in 100-mL LBG medium in 500-mL Erlenmeyer flasks. The medium was supplemented with chloramphenicol (15 g/ml) to maintain the plasmids. The initial optical density of the culture, measured at 600 nm, was adjusted to 0.001. The flasks were shaken at 150 rpm at 37 C. Bacterial cells were harvested at Abs 600 = 1.2 and washed once with cold M9-glucose. The cells were incubated with 10, 20, 50, 250, or 500 M [ 14 C] putrescine, and the reaction was terminated after 2, 4, 6, 8, and 10 min. The linearity between the uptake of [ 14 C] putrescine and the incubation time was confirmed. Supplementary tables Table S1 Statistically significant upregulated Gene Ontology (GO) terms in SK591 (pbelobac11 / yeef + ) compared with SK595 (pbelobac11 / yeef) (P < 0.05) GO term Term Number Fold-change P-value category of genes a average Z-score polyol metabolic 24 7.77E-15 1.47 7.77 anaerobic respiration 36 1.29E-09 0.94 6.07 nitrate assimilation 19 3.94E-07 1.08 5.07 glycerol metabolic 16 2.63E-06 1.09 4.70 solute:hydrogen symporter 51 1.62E-05 0.64 4.31 iron-sulfur cluster binding 133 2.10E-05 0.40 4.25 metal cluster binding 133 2.10E-05 0.40 4.25 ammonia-lyase 10 2.25E-05 1.38 4.24 solute:cation symporter 62 1.64E-04 0.51 3.77 1
phosphotransferase, alcohol group as acceptor amine catabolic substrate-specific transporter cell projection assembly substrate-specific cation-transporting ATPase two-component signal transduction system (phosphorelay) hexose transport carbohydrate-transporting ATPase active transmembrane transporter signal transmission polysaccharide localization molybdenum ion binding cellular carbohydrate metabolic flagellum organization signaling 151 3.44E-04 0.33 3.58 36 3.68E-04 0.55 3.56 342 4.52E-04 0.22 3.51 16 5.60E-04 0.80 3.45 289 6.09E-04 0.24 3.43 13 1.12E-03 0.94 3.26 82 2.65E-03 0.31 3.01 17 2.79E-03 0.68 2.99 11 3.51E-03 0.92 2.92 363 4.61E-03 0.18 2.83 262 5.50E-03 0.21 2.78 95 5.83E-03 0.27 2.76 12 7.54E-03 0.72 2.67 17 7.72E-03 0.68 2.66 267 8.82E-03 0.15 2.62 27 9.90E-03 0.46 2.58 97 1.24E-02 0.24 2.50 2
cellular component assembly at cellular level macromolecule localization secondary active cellular component assembly organic acid catabolic carboxylic acid catabolic cell projection organization phosphotransferase, nitrogenous group as acceptor D-gluconate metabolic membrane part monosaccharide cellular biogenic amine catabolic 64 1.32E-02 0.29 2.48 106 1.41E-02 0.23 2.46 131 1.41E-02 0.25 2.46 75 1.45E-02 0.27 2.44 65 1.48E-02 0.29 2.44 65 1.48E-02 0.29 2.44 28 1.64E-02 0.42 2.40 34 2.16E-02 0.43 2.30 10 2.24E-02 0.67 2.28 950 3.51E-02 0.10 2.11 22 3.68E-02 0.48 2.09 10 3.91E-02 0.61 2.06 a Number of genes represents the number of genes included in each pathway. Table S2 Statistically significant upregulated Gene Ontology (GO) terms in SK594 (pbelobac11-yeef + / yeef) compared with SK595 (pbelobac11 / yeef) 3
GO term polyol metabolic glycerol metabolic anaerobic respiration nitrate assimilation ammonia-lyase flagellum organization cation-transporting ATPase solute:hydrogen symporter cell projection organization ciliary or flagellar motility cell projection assembly solute:cation symporter amine catabolic iron-sulfur cluster binding metal cluster binding substrate-specific transporter substrate-specific Term category Number Fold-change P-value of genes a average Z-score 24 1.33E-15 1.30 7.99 16 3.92E-07 1.02 5.07 36 3.98E-07 0.69 5.07 19 5.41E-06 0.84 4.55 10 1.40E-05 1.20 4.34 27 3.07E-05 0.65 4.17 13 4.96E-05 0.99 4.06 51 6.39E-05 0.51 4.00 28 8.65E-05 0.61 3.93 30 1.17E-04 0.58 3.85 16 1.45E-04 0.77 3.80 62 5.04E-04 0.41 3.48 36 5.07E-04 0.48 3.48 133 1.71E-03 0.27 3.14 133 1.71E-03 0.27 3.14 342 2.00E-03 0.18 3.09 289 2.33E-03 0.19 3.04 4
cellular component assembly at cellular level phosphotransferase, alcohol group as acceptor cellular component assembly carbohydrate-transporting ATPase macromolecule localization methionine metabolic methionine biosynthetic hexose transport polysaccharide localization active transmembrane transporter organelle intracellular organelle cellular component organization at cellular level organic acid catabolic carboxylic acid catabolic flagellum part 64 5.38E-03 0.30 2.78 151 5.45E-03 0.23 2.78 75 5.99E-03 0.27 2.75 11 8.18E-03 0.71 2.64 106 1.03E-02 0.22 2.57 14 1.07E-02 0.56 2.55 14 1.07E-02 0.56 2.55 17 1.25E-02 0.50 2.50 12 1.42E-02 0.58 2.45 262 1.44E-02 0.17 2.45 363 2.15E-02 0.14 2.30 165 2.45E-02 0.20 2.25 165 2.45E-02 0.20 2.25 98 2.72E-02 0.20 2.21 65 2.77E-02 0.24 2.20 65 2.77E-02 0.24 2.20 23 2.79E-02 0.42 2.20 bacterial-type flagellum part 23 2.79E-02 0.42 2.20 5
cell projection part 23 2.79E-02 0.42 2.20 two-component signal transduction system (phosphorelay) 82 3.83E-02 0.20 2.07 cellular component biogenesis 147 5.00E-02 0.15 1.96 a Number of genes represents the number of genes included in each pathway. Table S3 Statistically significant downregulated Gene Ontology (GO) terms in SK591 (pbelobac11 / yeef + ) compared with SK595 (pbelobac11 / yeef) (P < 0.05) GO term Term Number Fold-change P-value category of genes a average Z-score arginine biosynthetic 13 8.64E-12-1.75-6.83 glutamine family amino acid biosynthetic 23 7.22E-09-1.11-5.79 organic acid biosynthetic 168 5.42E-06-0.32-4.55 amine biosynthetic 142 1.03E-05-0.34-4.41 cellular amino acid biosynthetic 134 1.08E-05-0.35-4.40 glutamine family amino acid metabolic 53 4.48E-05-0.51-4.08 response to stress 243 7.38E-05-0.23-3.96 small molecule biosynthetic 338 3.76E-04-0.17-3.56 response to oxidative stress 31 4.15E-04-0.58-3.53 cellular nitrogen compound 302 5.95E-04-0.18-3.43 6
biosynthetic cellular homeostasis monovalent inorganic cation homeostatic cellular response to oxidative stress response to chemical stimulus cellular response to stimulus oxidoreductase, acting on paired donors, with incorporation or reduction of molecular oxygen serine family amino acid biosynthetic cellular amino acid metabolic cellular response to chemical stimulus cellular amino acid and derivative metabolic purine nucleoside triphosphate biosynthetic purine ribonucleoside triphosphate biosynthetic ATP biosynthetic 45 2.00E-03-0.42-3.09 42 2.97E-03-0.43-2.97 51 4.28E-03-0.36-2.86 15 4.68E-03-0.67-2.83 186 6.20E-03-0.18-2.74 139 7.03E-03-0.21-2.70 19 9.43E-03-0.57-2.60 19 1.17E-02-0.53-2.52 229 1.28E-02-0.15-2.49 26 1.37E-02-0.44-2.47 260 1.37E-02-0.14-2.46 19 1.50E-02-0.51-2.43 19 1.50E-02-0.51-2.43 18 1.53E-02-0.52-2.42 cell redox homeostasis 21 1.65E-02-0.48-2.40 7
nucleoside triphosphate biosynthetic 22 2.01E-02-0.45-2.32 cellular response to reactive oxygen species DNA replication intracellular part cytoplasm regulation of biological quality coenzyme binding cellular biosynthetic transcription repressor transferase, transferring alkyl or aryl (other than methyl) groups ATPase, coupled to transmembrane movement of ions specific transcriptional repressor cell outer membrane biosynthetic peptide biosynthetic ribonucleoside triphosphate metabolic 12 2.35E-02-0.60-2.26 55 2.40E-02-0.28-2.26 885 2.58E-02-0.04-2.23 805 2.81E-02-0.04-2.20 98 3.03E-02-0.20-2.17 158 3.56E-02-0.13-2.10 1079 3.69E-02-0.05-2.09 75 3.90E-02-0.21-2.06 34 4.06E-02-0.32-2.05 31 4.08E-02-0.34-2.05 35 4.16E-02-0.31-2.04 84 4.16E-02-0.18-2.04 1102 4.17E-02-0.05-2.04 10 4.21E-02-0.59-2.03 22 4.23E-02-0.39-2.03 membrane fraction 10 4.59E-02-0.57-2.00 8
insoluble fraction 10 4.59E-02-0.57-2.00 cellular chemical homeostasis 24 4.67E-02-0.37-1.99 ion homeostasis 21 4.85E-02-0.39-1.97 a Number of genes represents the number of genes included in each pathway. Table S4 Statistically significant downregulated Gene Ontology (GO) terms in SK594 (pbelobac11-yeef + / yeef) compared with SK595 (pbelobac11 / yeef) GO term Term Number Fold-change P-value category of genes a average Z-score arginine biosynthetic 13 4.38E-12-1.48-6.92 glutamine family amino acid biosynthetic 23 8.03E-09-0.92-5.77 response to stress 243 2.28E-04-0.16-3.69 amine biosynthetic 142 3.69E-04-0.21-3.56 organic acid biosynthetic 168 4.79E-04-0.19-3.49 cellular amino acid biosynthetic 134 6.45E-04-0.21-3.41 response to oxidative stress 31 1.09E-03-0.44-3.27 glutamine family amino acid metabolic 53 1.31E-03-0.32-3.21 monovalent inorganic 42 1.52E-03-0.38-3.17 9
cation transmembrane transporter oxidoreductase, acting on paired donors, with incorporation or reduction of molecular oxygen coenzyme binding cellular response to oxidative stress response to chemical stimulus cellular nitrogen compound biosynthetic cellular response to stimulus homeostatic cellular homeostasis small molecule biosynthetic serine family amino acid biosynthetic oxidoreductase, acting on single donors with incorporation of molecular oxygen oxidoreductase, acting on single donors with incorporation of molecular oxygen, incorporation of two atoms of oxygen 19 3.95E-03-0.52-2.88 158 9.08E-03-0.14-2.61 15 9.50E-03-0.50-2.59 186 1.03E-02-0.12-2.57 302 1.25E-02-0.09-2.50 139 1.30E-02-0.14-2.48 51 1.31E-02-0.25-2.48 45 1.38E-02-0.26-2.46 338 1.51E-02-0.08-2.43 19 2.64E-02-0.38-2.22 15 2.74E-02-0.45-2.21 15 2.74E-02-0.45-2.21 10
cellular response to chemical stimulus transcription repressor cellular aldehyde metabolic polysaccharide biosynthetic specific transcriptional repressor membrane fraction insoluble fraction cell redox homeostasis copper ion binding external encapsulating structure part 26 2.78E-02-0.31-2.20 75 2.84E-02-0.18-2.19 17 3.15E-02-0.39-2.15 144 3.40E-02-0.11-2.12 35 3.72E-02-0.26-2.08 10 4.29E-02-0.46-2.02 10 4.29E-02-0.46-2.02 21 4.31E-02-0.32-2.02 13 4.69E-02-0.43-1.99 141 4.72E-02-0.08-1.98 a Number of genes represents the number of genes included in each pathway. 11
Fig. S1 2 2 1/v (nm mol/min/mg) -1 1.6 1.2 0.8 0.4 0 0 y = 16.579x + 0.1071 R² = 0.9993-0.03 0 0.03 0.06 0.09 0.12 1/s ( M) -1 12