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| Time-dependent concentration profile of extracellular detected amino acids and fold-changes of secreted metabolites after growth of S. solfataricus P2 on caseinhydrolysate. The Initial concentration of every amino acid at time point zero (except glutamate & pyroglutamate) was adapted from Neumann-Schaal et. al. (2015). | ||||||||||||||||||||||||||||||||||||
| Glutamate and pyroglutamate were quantified via HPLC. | ||||||||||||||||||||||||||||||||||||
| Fold-changes of secreted metabolites were calculated compared to the time point of their first appearance in the media (indicated by a value of “1.00”). For detection of short chain organic acids only five sampling time points were analysed. Values represent the average of three independent experiments. | ||||||||||||||||||||||||||||||||||||
| Metabolites | Amino acid concentration [mg∙l-1] | |||||||||||||||||||||||||||||||||||
| 0 h | 21 h | 45 h | 66 h | 77 h | 89 h | 101 h | References: | |||||||||||||||||||||||||||||
| Alanine | 160.00000000000003 | ± | 6.881228093254414 | 167.9904847230737 | ± | 17.832084847299004 | 152.6978435843025 | ± | 4.084520681751968 | 136.27467596942472 | ± | 7.969484434322831 | 141.72438955124238 | ± | 7.3006530047756915 | 130.67011504093514 | ± | 3.48942817614711 | 129.2480190081339 | ± | 2.2906106595815787 | Neumann-Schaal M, Hofmann JD, Will SE & Schomburg D (2015) Time-resolved amino acid uptake of Clostridium difficile 630Δerm and concomitant fermentation product and toxin formation. BMC Microbiol. 15: Available at: http://www.biomedcentral.com/1471-2180/15/281 | ||||||||||||||
| Arginine | 140 | ± | 28.606233837524275 | 169.59594033271466 | ± | 23.637911380552953 | 151.57551128896284 | ± | 23.43215718865023 | 182.78737143309874 | ± | 24.481659264331597 | 293.5271140418977 | ± | 20.750250005270146 | 266.2919731463383 | ± | 17.373303917311134 | 250.45178853015236 | ± | 44.05513038492533 | |||||||||||||||
| Aspartate | 270.00000000000006 | ± | 5.31345678122099 | 280.4128341820452 | ± | 11.03358128837849 | 260.5075362096365 | ± | 7.422339559148695 | 257.80306884484736 | ± | 17.741154428951255 | 236.98515290257157 | ± | 11.550955444809809 | 243.2440521554225 | ± | 17.023572946030274 | 221.24344662497012 | ± | 11.312608406429048 | |||||||||||||||
| Glutamate | 821.4665351433 | ± | 12.1223203929073 | 679.841598199442 | ± | 18.1436084619454 | 476 | ± | 21 | 321 | ± | 6 | 270 | ± | 4 | 253 | ± | 11 | 275 | ± | 4 | |||||||||||||||
| Glycine | 50 | ± | 2.098010481590222 | 51.32849021477633 | ± | 2.644668729553069 | 47.731333757339286 | ± | 1.3949439631230287 | 45.52253591146408 | ± | 2.3714789157382237 | 43.79937734509527 | ± | 0.8335903462780071 | 43.05782077237285 | ± | 1.1661687556487126 | 41.99927528398161 | ± | 1.5084370336453974 | |||||||||||||||
| Histidine | 90 | ± | 12.500136116226736 | 100.14570777680835 | ± | 10.122261766041865 | 93.22093667341653 | ± | 10.082432584268695 | 112.38399914711017 | ± | 12.18687225013122 | 125.57259100450267 | ± | 6.831667648495093 | 136.47067664028273 | ± | 4.085769811430832 | 131.86232989753682 | ± | 12.187062308560394 | |||||||||||||||
| Isoleucine | 240 | ± | 20.128715675066182 | 221.7161838796816 | ± | 17.429115524817338 | 234.7751465687295 | ± | 10.458021492396405 | 218.60228585356944 | ± | 2.1499663155984416 | 208.77449439183104 | ± | 5.972264345065796 | 188.81281954074942 | ± | 8.663973543177988 | 169.09942467721604 | ± | 4.8862530218027205 | |||||||||||||||
| Leucine | 380 | ± | 9.61181364822445 | 399.8485463736368 | ± | 19.767406974310262 | 355.00085271330727 | ± | 13.213245481706764 | 311.07966348388635 | ± | 2.761445184416518 | 282.24153878845846 | ± | 30.281219997914242 | 229.8433003230009 | ± | 17.989641346560955 | 192.5926068287889 | ± | 15.191100093452643 | |||||||||||||||
| Lysine | 340 | ± | 6.326321686643842 | 339.94763466917226 | ± | 4.261925714028086 | 361.08901768314007 | ± | 1.858861311192359 | 389.3459788407459 | ± | 15.16129747225218 | 384.53189681389404 | ± | 2.65390953368356 | 425.06970119757904 | ± | 21.489638659669055 | 450.85127781900724 | ± | 7.10771535708484 | |||||||||||||||
| Methionine | 119.99999999999999 | ± | 8.125043518931356 | 125.92844303758568 | ± | 21.3113379884199 | 97.13987654044888 | ± | 2.059281331449405 | 87.9790751901548 | ± | 3.319469743373236 | 76.64930304712223 | ± | 1.2524826297792515 | 73.05691879580475 | ± | 6.995153021424486 | 54.43205936839782 | ± | 8.450904870315457 | |||||||||||||||
| Phenylalanine | 200.00000000000003 | ± | 0.415339771072415 | 197.6083025387518 | ± | 1.132054443788466 | 204.8362979586142 | ± | 6.107976064935889 | 199.46741189029558 | ± | 5.510484379499272 | 186.73107938528915 | ± | 11.434905881775121 | 159.88806673000659 | ± | 7.146024308926436 | 135.38886439742285 | ± | 16.215184404937716 | |||||||||||||||
| Proline | 360 | ± | 6.168814216156419 | 387.746513251376 | ± | 17.575690266403022 | 389.9641429594319 | ± | 18.877687546536222 | 416.8802886698649 | ± | 15.459967510273641 | 424.80045890475805 | ± | 4.778212792697188 | 445.8649294113687 | ± | 1.395763856112424 | 450.6365477967922 | ± | 12.936518730097799 | |||||||||||||||
| Pyroglutamate | 0 | ± | 11.4786141419509 | 23.6474283456394 | ± | 17.3334144494667 | 145 | ± | 14 | 188 | ± | 11 | 180 | ± | 6 | 164 | ± | 13 | 117 | ± | 7 | |||||||||||||||
| Serine | 250 | ± | 3.810623651753096 | 256.2810053155547 | ± | 6.280531413750752 | 254.9684173125364 | ± | 4.229287828597417 | 255.7413744544335 | ± | 13.064749398030791 | 247.242958330334 | ± | 10.313959253310816 | 240.01494252372194 | ± | 3.10084244997565 | 241.11085607138133 | ± | 6.538157906163563 | |||||||||||||||
| Threonine | 210 | ± | 12.981699079722187 | 222.03481886275017 | ± | 33.631189875829065 | 188.55612388555414 | ± | 4.318558811943741 | 185.57248383665979 | ± | 6.26912769119231 | 177.30857995964553 | ± | 4.66567198635362 | 172.69095896074805 | ± | 6.5052453901536085 | 168.03824184001422 | ± | 4.242144473301449 | |||||||||||||||
| Tyrosine | 239.99999999999997 | ± | 6.062532937301132 | 248.25215464369094 | ± | 1.6834892574164984 | 238.17238457381154 | ± | 7.509818752896757 | 232.7437978134993 | ± | 7.91955534218896 | 225.4720815364394 | ± | 2.952328003923748 | 213.31748683036355 | ± | 8.357735189356617 | 204.22544085255322 | ± | 6.246520432024575 | |||||||||||||||
| Valine | 270.00000000000006 | ± | 5.998743898183086 | 286.27441852109274 | ± | 17.20344741182678 | 292.4296709107175 | ± | 9.36980402232918 | 306.55727429261134 | ± | 8.62693061612567 | 304.4214204524945 | ± | 5.3677589995033035 | 300.41674649536697 | ± | 5.167053010316502 | 288.47512492543075 | ± | 3.7318584380460504 | |||||||||||||||
| Others | Fold change in abundance | |||||||||||||||||||||||||||||||||||
| 2-Hydroxyphenylacetate | 1 | 0.03 | 1.78 | ± | 0.12 | 1.7 | ± | 0.06 | 2.05 | ± | 0.39 | |||||||||||||||||||||||||
| 3-Hydroxyisovaleric acid | 1 | ± | 0.21 | 2.22 | 0.65 | 2.97 | ± | 0.57 | 4.74 | ± | 0.33 | 6.04 | ± | 0.76 | ||||||||||||||||||||||
| 3-Phosphoglycerate | 1 | ± | 0.2 | 1.9 | 0.21 | 3.32 | ± | 0.32 | 3.92 | ± | 0.44 | 4.13 | ± | 0.71 | ||||||||||||||||||||||
| 4-Hydroxybenzoate | 1 | ± | 0.08 | 2.07 | 0.14 | 2.46 | ± | 0.12 | 3.01 | ± | 0.13 | 3.41 | ± | 0.19 | ||||||||||||||||||||||
| 4-Hydroxyphenylacetate | 1 | ± | 0.14 | 5.78 | ± | 0.49 | 12.68 | 1.3 | 16.93 | ± | 1.02 | 22.99 | ± | 2.27 | 27.42 | ± | 2.6 | |||||||||||||||||||
| Glycerol-3-phosphate | 1 | ± | 0.1 | 2.66 | ± | 0.22 | 4.23 | 0.38 | 4.61 | ± | 0.23 | 4.93 | ± | 0.46 | 5 | ± | 0.3 | |||||||||||||||||||
| Glycylglycylglycine | 1 | ± | 0.21 | 1.22 | ± | 0.12 | 1.09 | ± | 0.13 | 0.92 | 0.15 | 1.02 | ± | 0.01 | 0.94 | ± | 0.13 | 0.79 | ± | 0.16 | ||||||||||||||||
| Homoserine | 1 | ± | 0.21 | 1.76 | ± | 0.22 | ||||||||||||||||||||||||||||||
| Ornithine | 1 | ± | 0.08 | 1.3 | ± | 0.09 | 1.03 | ± | 0.15 | 1.22 | ± | 0.05 | 1.69 | ± | 0.14 | 1.39 | ± | 0.19 | 1.28 | ± | 0.16 | |||||||||||||||
| Pipecolate | 1 | ± | 0.12 | 2.66 | 0.4 | 3.69 | ± | 0.34 | 5.24 | ± | 0.53 | 6.03 | ± | 0.39 | ||||||||||||||||||||||
| Trehalose | 1 | ± | 0.19 | 10 | 4.54 | 7.74 | ± | 4.01 | 22.32 | ± | 6.42 | 23.59 | ± | 2.83 | ||||||||||||||||||||||
| Urea | 1 | ± | 0.1 | 1.34 | 0.07 | 1.53 | ± | 0.29 | 1.69 | ± | 0.22 | 1.37 | ± | 0.22 | ||||||||||||||||||||||
| Short chain organic acids | ||||||||||||||||||||||||||||||||||||
| 2-Methylbutanoate | 1 | ± | 0.23 | 2.39 | ± | 0.4 | 6.37 | ± | 0.85 | 6.83 | ± | 0.14 | 6.3 | ± | 1.14 | |||||||||||||||||||||
| 2-Methylcrotonate | 1 | ± | 0.16 | 3.69 | ± | 0.42 | 13.41 | ± | 2.8 | 17.16 | ± | 2.66 | 17.42 | ± | 0.71 | |||||||||||||||||||||
| 3-Methylcrotonate | 1 | ± | 0.08 | 2.22 | ± | 0.28 | 7.85 | ± | 3.16 | 8.53 | ± | 1.04 | 9.6 | ± | 1.65 | |||||||||||||||||||||
| Isobutanoate | 1 | ± | 0.52 | 2.89 | ± | 2.2 | 5 | ± | 0.91 | 6.9 | ± | 0.72 | 5.85 | ± | 0.88 | |||||||||||||||||||||
| Isovalerate | 1 | ± | 0.09 | 5.53 | ± | 0.6 | 17.11 | ± | 2.56 | 22.29 | ± | 1.87 | 21.37 | ± | 2.31 | |||||||||||||||||||||
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| Time dependent concentration profile of detected amino acids in a non-inoculated control culture, to investigate stability of compounds under cultivation conditions. The Initial concentration of every amino acid at time point zero (except glutamate & pyroglutamate) was adapted from Neumann-Schaal et. al. (2015). | |||||||||||||||||||||||||||||
| Glutamate and pyroglutamate were quantified via HPLC. | |||||||||||||||||||||||||||||
| For glycylglycylglycine and ornithine absolute concentrations could not be calculated, therefore the fold-change compared to time point zero is given. | |||||||||||||||||||||||||||||
| Values represent the average of two technical replicates. | |||||||||||||||||||||||||||||
| Note that neither volatile fatty acids nor other compounds than amino acids were found in the control. | |||||||||||||||||||||||||||||
| Metabolites | Amino acid concentration [mg∙ml-1] | ||||||||||||||||||||||||||||
| 0 h | 21 h | 45 h | 66 h | 77 h | 89 h | 101 h | References: | ||||||||||||||||||||||
| Amino acids | Neumann-Schaal M, Hofmann JD, Will SE & Schomburg D (2015) Time-resolved amino acid uptake of Clostridium difficile 630Δerm and concomitant fermentation product and toxin formation. BMC Microbiol. 15: Available at: http://www.biomedcentral.com/1471-2180/15/281 | ||||||||||||||||||||||||||||
| Alanine | 160 | ± | 3.27191726780034 | 160.39858725742036 | ± | 9.310421960657509 | 161.85751468923007 | ± | 3.5331757392202197 | 164.9976219203087 | ± | 2.8965145045993568 | 172.59356902222848 | ± | 0.7301999766724905 | 170.8021411421057 | ± | 4.795220381850258 | 187.35386357684763 | ± | 17.559432389671887 | ||||||||
| Arginine | 140 | ± | 1.6664845712241039 | 127.58410637895605 | ± | 29.63149170512144 | 159.97107951132233 | ± | 7.4171934623549 | 124.80898714725431 | ± | 5.952774406980075 | 195.49441835545178 | ± | 9.271774974727768 | 166.76653977429908 | ± | 6.497686940735158 | 149.04756661035415 | ± | 16.1748935807662 | ||||||||
| Aspartate | 270 | ± | 0.22712828026476062 | 255.5393835104091 | ± | 20.2825822046744 | 278.60041393326253 | ± | 7.549375722644592 | 255.0701656508327 | ± | 0.15380022131317386 | 286.61104338414503 | ± | 7.495663954303049 | 281.4672029593636 | ± | 1.4081468420550771 | 287.4138006937003 | ± | 4.1975872345607765 | ||||||||
| Glutamate | 813 | ± | 12 | 756 | ± | 13 | 715 | ± | 15 | 653 | ± | 5 | 564 | ± | 3 | 469 | ± | 3 | 419 | ± | 11 | ||||||||
| Glycine | 50 | ± | 0.7613064160611265 | 50.65994009309705 | ± | 3.4273114819711097 | 52.64952301166585 | ± | 0.7523321002024604 | 51.81636285908343 | ± | 0.2404500777172424 | 55.10646551287211 | ± | 0.08712388712669886 | 53.25894106266598 | ± | 0.5809119261676615 | 55.519777906272665 | ± | 1.196779971558684 | ||||||||
| Histidine | 90 | ± | 1.1516270888887876 | 78.78104713968736 | ± | 13.90145188895471 | 84.27833339281878 | ± | 6.3071915920272 | 71.20378921192068 | ± | 4.751986358617884 | 81.89959812376495 | ± | 10.349211512335295 | 85.26182058854494 | ± | 1.0739699309455661 | 74.24816574533466 | ± | 15.726531870133615 | ||||||||
| Isoleucine | 240 | ± | 2.431983100912866 | 247.8196584714675 | ± | 9.42666637045332 | 252.87910810559913 | ± | 14.082782737417222 | 255.74108896726406 | ± | 11.209249237345013 | 251.5202145387305 | ± | 8.71456476598796 | 257.05235947799736 | ± | 9.480860631287218 | 269.880913587243 | ± | 31.901139894269573 | ||||||||
| Leucine | 380 | ± | 9.093573303475925 | 359.9235219456074 | ± | 11.353094100753315 | 384.69966493626316 | ± | 19.70285366720812 | 375.1314977797677 | ± | 3.0691998068408517 | 377.8705726773993 | ± | 1.1018686598566985 | 381.9780137083726 | ± | 2.3685986398108705 | 401.9908423570388 | ± | 10.052176191873889 | ||||||||
| Lysine | 340 | ± | 1.034614860773675 | 343.60854845519134 | ± | 1.8693662595561507 | 336.95743049668124 | ± | 0.8774054928541676 | 356.30943033491155 | ± | 9.709173124543426 | 336.5036371703973 | ± | 0.7181914881191076 | 343.14984694154435 | ± | 2.352254600604939 | 351.84865028511797 | ± | 15.687114911956945 | ||||||||
| Methionine | 120 | ± | 3.759316861401942 | 111.59032507714073 | ± | 7.752588850383391 | 120.39637778768486 | ± | 1.7470809574273338 | 105.61270527632345 | ± | 1.5476513848664633 | 124.04701354081516 | ± | 10.631367232873227 | 118.24617235179122 | ± | 0.5947502129925253 | 132.05548415647692 | ± | 19.9051753155092 | ||||||||
| Phenylalanine | 200 | ± | 2.600580680686207 | 194.96220752695532 | ± | 2.565050788519631 | 194.8255470130046 | ± | 5.71204816810147 | 193.66446186281297 | ± | 0.21926633357003603 | 188.9586969965307 | ± | 0.6325521210761924 | 189.08911190306756 | ± | 1.531052160738673 | 186.28976140679376 | ± | 0.12540439518095997 | ||||||||
| Proline | 360 | ± | 1.213810651057944 | 346.5816626953414 | ± | 7.847387592152955 | 370.4572176544668 | ± | 17.70413679063146 | 369.5656596728953 | ± | 4.833842988824444 | 362.70274420907157 | ± | 0.8845996967872803 | 369.7748329984605 | ± | 4.774189116080444 | 388.7485067735637 | ± | 0.3925616603731171 | ||||||||
| Pyroglutamate | 0 | ± | 3 | 25 | ± | 15 | 70 | ± | 13 | 107 | ± | 12 | 206 | ± | 2 | 281 | ± | 5 | 318 | ± | 15 | ||||||||
| Serine | 250 | ± | 3.103051152802013 | 248.3911040815742 | ± | 5.841893634010814 | 262.0879065601382 | ± | 12.555185906035854 | 259.7164777759091 | ± | 2.4146188747695003 | 261.7307674616085 | ± | 2.909167486169531 | 262.49600365254787 | ± | 0.7932210608588763 | 268.12106827701484 | ± | 1.8421851956720592 | ||||||||
| Threonine | 210 | ± | 1.758140561469365 | 208.4479217645681 | ± | 7.698367128970491 | 218.96164520395052 | ± | 6.009444044109829 | 215.38805889327728 | ± | 1.4376782986972128 | 229.9212427476226 | ± | 8.900301832795833 | 224.57925683998323 | ± | 1.3950164789391168 | 273.98864532502853 | ± | 69.30143384153415 | ||||||||
| Tyrosine | 240 | ± | 1.7468954048425545 | 233.170764844196 | ± | 9.537784400293127 | 242.3668051509619 | ± | 1.953784722704232 | 238.77661244762763 | ± | 1.176388120997403 | 243.18938932423134 | ± | 0.8067725421760146 | 241.8826118928191 | ± | 0.5517411644078168 | 242.8931016383067 | ± | 2.9058136911040955 | ||||||||
| Valine | 270 | ± | 3.9313477227649156 | 262.0018037465443 | ± | 3.3306184168211015 | 274.257546822633 | ± | 7.680099119346837 | 273.114516674167 | ± | 1.8579806647474784 | 272.6506362294193 | ± | 0.9779690852742124 | 273.94853937447243 | ± | 6.125833864878838 | 291.68874660021515 | ± | 16.229249402354952 | ||||||||
| Others | |||||||||||||||||||||||||||||
| Glycylglycylglycine | 1 | ± | 0.07 | 0.93 | ± | 0.19 | 1.1 | ± | 0.06 | 0.98 | ± | 0.03 | 1.16 | ± | 0.06 | 1.13 | ± | 0.05 | 0.96 | ± | 0.26 | ||||||||
| Ornithine | 1 | ± | 0.1 | 0.87 | ± | 0.07 | 1.14 | ± | 0.09 | 0.81 | ± | 0.05 | 1.16 | ± | 0.08 | 1.11 | ± | 0.03 | 0.98 | ± | 0.17 | ||||||||
