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The \ key aspects and approaches of each phase of the design build test learn cycle are presented. Thecycle starts with a defined eng\ ineering objective that determines th:83)/F 4/H/N/Rect[315.392 675.855 319.802 663.444]/Subtype/Link/Type/Annot>> endobj 4 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:12. Wheelwright, S. C. & Clark, K. B. Revolutionizing Product Development: Quantum L\ eaps in Speed, Efficiency, and Quality \(The Free Press, 1992\).\r:84)/F 4/H/N/Rect[334.719 664.855 339.761 652.444]/Subtype/Link/Type/Annot>> endobj 5 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:13. Nielsen, J. & Keasling, J. D. Engineering cellular metabolism. Cell 164, 1\ 185 1197 \(2016\). \nThis review highlights experiences, success stories and challenges associated with implementing the DBTL cy\ cle for metabolic engineering.\r:85)/F 4/H/N/Rect[225.359 653.855 230.553 641.444]/Subtype/Link/Type/Annot>> endobj 6 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:14. Blank, S. & Dorf, B. The Startup Owner s Manual: \nThe Step- by-Step Guide for Bu\ ilding a Great Company \(K&S Ranch, 2012\).\r:86)/F 4/H/N/Rect[199.574 642.855 204.67 630.444]/Subtype/Link/Type/Annot>> endobj 7 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:15. Jansson, J. K. & Hofmockel, K. S. The soil microbiome  from metagenomics to m\ etaphenomics. Curr. Opin. Microbiol. 43, 162 168 \(2018\).\r:87)/F 4/H/N/Rect[547.618 587.855 552.756 575.444]/Subtype/Link/Type/Annot>> endobj 8 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:Fig. 2 | Top- down and bottom- up approaches to design microbiomes. The left\ panel illustrates a bottom- up design workflow starting from isolates. Physiological characterization of individual organisms i\ s performed, and metabolic modelling :88)/F 4/H/N/Rect[369.702 576.855 374.167 564.444]/Subtype/Link/Type/Annot>> endobj 9 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:16. Briones, A. & Raskin, L. Diversity and dynamics of microbial communities in en\ gineered environments and their implications for process stability. Curr. Opin. Biotechnol. 14, 270 276 \(2003\).\r:89)/F 4/H/N/Rect[528.219 510.855 533.314 498.444]/Subtype/Link/Type/Annot>> endobj 10 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:17. Verstraete, W. etal. Microbial resource management: the road to go for enviro\ nmental biotechnology. \nEng. Life Sci. 7, 117 126 \(2007\).\r:90)/F 4/H/N/Rect[509.859 499.855 515.033 487.444]/Subtype/Link/Type/Annot>> endobj 11 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:18. Moralejo- Grate, H., Mar Atusalihat, E., Kleerebezem, R. & Van Loosdrecht, M.\ C. M. Microbial community engineering for biopolymer production from \nglycerol. Appl. Microbiol. Biotechnol. 92, 631 639 \(\ 2011\).\r:91)/F 4/H/N/Rect[455.969 488.855 461.064 476.444]/Subtype/Link/Type/Annot>> endobj 12 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:19. Nielsen, P. H. etal. A conceptual ecosystem model \nof microbial communities i\ n enhanced biological phosphorus removal plants. Water Res. 44, 5070 5088 \(2010\).\r:92)/F 4/H/N/Rect[538.927 422.855 544.16 410.444]/Subtype/Link/Type/Annot>> endobj 13 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:20. Winkler, M.-K. H. etal. An integrative review of granular sludge for the biol\ ogical removal of nutrients and recalcitrant organic matter from wastewater. Chem. Eng. J. 336, 489 502 \(2018\).\r:93)/F 4/H/N/Rect[545.425 422.855 550.658 410.444]/Subtype/Link/Type/Annot>> endobj 14 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:21. Henze, M., Gujer, W., Mino, T. & Van Loosdrecht, M. C. M. Activated Sludge Model\ s ASM1, ASM2, ASM2d and ASM3 \(IWA Publishing, 2000\).\r:94)/F 4/H/N/Rect[420.607 301.855 425.901 289.444]/Subtype/Link/Type/Annot>> endobj 15 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:23. Muoz- Tamayo, R., Giger- Reverdin, S. & Sauvant, D. Mechanistic modelling of \ invitro fermentation and methane production by rumen microbiota. Anim. Feed. Sci. Technol. 220, 1 21 \(2016\).\r:95)/F 4/H/N/Rect[428.762 301.855 434.055 289.444]/Subtype/Link/Type/Annot>> endobj 16 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:24. Picioreanu, C., Kreft, J. & Loosdrecht, M. C. M. \nVan. Particle- based multidi\ mensional multispecies biofilm model. Appl. Environ. Microbiol. 70, 3024 3040 \(2004\).\r:96)/F 4/H/N/Rect[410.882 246.855 415.978 234.444]/Subtype/Link/Type/Annot>> endobj 17 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:25. Yunjie Ma, Carlos Domingo- Felez, Benedek Gy. Plosz & B. F. S. Intermittent ae\ ration suppresses nitrite- oxidizing bacteria in membrane- aerated biofilms: \na model- based explanation. Environ. Sci. Techno\ l. 51, 6146 6155 \(2017\).\r:97)/F 4/H/N/Rect[417.174 246.855 422.269 234.444]/Subtype/Link/Type/Annot>> endobj 18 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:21. Henze, M., Gujer, W., Mino, T. & Van Loosdrecht, M. C. M. Activated Sludge Mode\ ls ASM1, ASM2, ASM2d and ASM3 \(IWA Publishing, 2000\).\r:98)/F 4/H/N/Rect[531.443 191.855 536.637 179.444]/Subtype/Link/Type/Annot>> endobj 19 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:4. Lffler, F. E. & Edwards, E. A. Harnessing microbial activities for environmen\ tal cleanup. Curr. Opin. Biotechnol. 17, 274 284 \(2006\).\r:99)/F 4/H/N/Rect[410.854 180.855 413.48 168.444]/Subtype/Link/Type/Annot>> endobj 20 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:26. Nobu, M. K. etal. Microbial dark matter ecogenomics reveals complex synergist\ ic networks in a methanogenic bioreactor. ISME J. 9, 1710 1722 \(2015\).\r:100)/F 4/H/N/Rect[448.037 158.855 453.227 146.444]/Subtype/Link/Type/Annot>> endobj 21 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:27. Rotaru, A. E. etal. A new model for electron flow during anaerobic digestion:\ direct interspecies electron transfer to Methanosaeta for the reduction \nof carbon dioxide to methane. Energy Environ. Sci. \ 7, 408 415 \(2014\).\r:101)/F 4/H/N/Rect[478.824 125.855 483.891 113.444]/Subtype/Link/Type/Annot>> endobj 22 0 obj <>/ExtGState<>/Font<>/ProcSet[/PDF/Text]/XObject<>>> endobj 23 0 obj <>stream x}Y#Gr{# +y2$E-]y`P@VUXc'4u#3Q@7ufӕ̈?>?B泯G{WH۽7_ܾ-D [|,nTṇ]¿15U%:KSU&:2xiqZ̳Ov쵲5*noV*v2c"X/MWeHjo8#d~WJ+PJOSFϝꩍU}[(&#kZh7kj#/Yf4DiC^tZE5Wj.qG]w|Ɯ56\uГ_V#;zxZ 5ٷir SN+e⓷Ce1}ِ~{趤,[RQ)J ( I6R1!hz7_LcF:dOxܩhp_[VvLtE'{fA\̐*gu#!TY|.jn6B"{)Uo~d!p;8}$OZ^wݗOn'[01G-CX^=h&d@6#&憁vS[~Ӳb`z 93~يQ,̎hǢO vΟ/{l#䨂)״؆!.z1T4[ܥv-"h(ՋljU ܌Q9viDX3P|P'1%r|gIL9M~MT 'йIpՅ # cW(Ӻ׍?oIӊ{j23W#r:xl?qdvQ|Zg"f|u(íOэ&M vJ6IDn J*xA{NMx܄%]c7h!$N,I=xaF<ӌtMe> a`tfJ;& p,t*{kux60 u4P)Ѷ~ܳ lh w]8e2l?RR\͘f'G.OV<*dB۠tLtB:Dwq;ș{ zGnD{h :Jϖi:]׸;n> -f1@{v\}!8-M{UVmՆ- PP;ZSFagP90nG؊)p9)%=.d`N+΄jY֞EazfnbN0S :o[3/;ng#*k?מۀe.ıܥ0 .@ǂd+^*z{&ׂõ|`΄mٮ>X>Lj6W` ,\<}ji|ŝz[dS8fL fW8oўork]@ 2ؾsXXb^]@,/e^,ԅ !fmIm`Y_sU_ZeESt9%}h:D?1p;g#rKd#{Eo~kǏz'ݴw Q'*,Us6=c]qci@`1wB >3֬^TqM)a%d;!$ s#-ϰf X}qJeW-Jxˣb'cf'agC4Kv ^| Ozm3%k-OʶАdЍj0~m:5S4Y)H hw.7lF" 3[L{dYBd4<ឋqݎ8GgFq.P\T@wQ r$caϯ4ͧǬ8)lٳ1+FG!V-1o3ώ=*qيm]nUTϙj1ĊޘOE7|åJsMhmaj.+ g7#oI&I׽XG?׉F]069@kLY+m17yE 멷c*N'd!+ۺҊ}s//)=(2:#hSԍ;mc]cbHf#SLsj4o5=4aX-AYu!w̥z\8z{Z+ڗg%B$~l] hDR%x&ʾ{.JbJDԏG'YKg!YΜb<2kEG0xAX [4?HtNSR.B ߱x;1iAO.'J p*+ʗ#yfs$-8p C2 FYwV;dM[ctBF~Įbŧh;'mՃ;D m2 3"B3p'P/<)&̓Fl|Ie$&DM8F6 ؽ LgE@Uam뎡^fmLssgynM_cµR ؉d|)2/}>琿5D:T~Ip"$L3c( ю_ݬCa g_C e-' 7 t ;9\0ѮJZ~'p1K2c}BB:\1f;W!JjM¹Z);5Qo.;yv;l1ӯ,(V60z~zW|߾Yh"jhb g6 ~iYm pufRh*>y("|(+YD֓U~m#Y(w6̃%bߠX9%@<`-'d}xqp4y8¡*G o+kEgP:A"&DOwC=ٕp_d„ x'%ܴÚ<k\fqyvyPAWFZe;0[hjH(^5 ߲с> =Tj})*I_Qu\wH&"´Jud5$2TG,݆T&̻yaD|{\5|*>³؈݃J$,EFNd K_-Ĩe#]Ώy1y Rb/b?D=#/ؘ4-=ܱ&ڪy'_w\<\4H +e4qiE]OaB}sya tf'U@?VyY9PS8AwlwGrk}&q9:.g 85%6]R:ϔgŕuչʫ8g?60mgY&AOMsܖx3/OJN0V΋zahf @>*OoK :M%qNX&?{Aq`h|5(#$\<-@C/).QJ%nC~Sչ23rƔD|H|vey˹+t1_`)H| 5| MdgigM |{h(opz}.d¦V8]Y9 >mq Qo霨{?|0z Q0K <19LU޼ w(Q"{i6Cm@Sd>b@l}/ ^)Clci'j+WEfd&AUsEV8`X$砘&3%EU|ewƐ <d4Ӱ_GHI#yŽ0"{U\Mb5 Va̐CKka3-`mZd̸%XX-:R҆b`ٍ]Gl*>U@z$4vXfnn.k%'<+_eUeU'Z*hUY*$B~τXAq ݹ]hz3Ox[m`G3[>u.YzE\ &.Qq"!=Dͳ9WՙH^(%ӀRʏ3*EiD)蓝A,W:EF`d\$;lrN<;Gi@hggQZѠ2YL͗\C?J+ra/?#U seQ s#7M)s\^ڳ( Vef(."P7? ~O<)a+#{6*'r}B4U,TXWKfXYӺHkҸ˸aD x.I覾_j ~e8]hƘ"&pW}5 x@li$ʄ)5y|u-e䓹ayEC܀ŁRݪWT䫲5FHHs>:Ҫdq*Һ&s[8jpq6T.!l-p BMjebhE;C)6m%v?כ^'Yl00΄\ 7=ZYǡh3qHtt=s<5P˲j_S jŔyd UAGYHa#u>.zg2MҶd](qZq9 "6\Vkax3H{yTXE)+,26B)ݜ Z'M|ɢ#W0 7ND't0oFmv:ieC:xKccS:Yz؃ q t5Z[ʅZXHZk !SgDplkbVh˜Dzg,D7c&'z8U7V1~(QH*^/Q)ǁ9&a>wJ5=5dѠIEe%Ϥx6]z^Yb+B4[]{A:oK yq @,]D PS#Z &,TsY &y+.DZITcBɰV(0t"muO ŪЅƦ46tu?):8c;eş;#"s^M0>|^#i(/6 V>oÅ@t$lFq&'Xp7X:$~jGգa+2^r'Ul<擆V'Ex[^Q+\-6WO׿uDgH?s_ig! ai%?h/G2B2FK%OD5`'Ӈf.:Յ`e(W;^#ˆ42(0ҥ+SٹvXMp/,[ҵV` 傫Z))eV{vRx)M r_pVY*7% V u/A+ )?>E~(TI&+^⫙Ɋ":o&{B s K'Q\/l!I^or%Byys1vMO.4?M&_Ik|`5LӢJաy7P={b=hz~ }$Z t,@NUqTfZn͛YRO SAX/`1i]tF @YRzu7=76T? (rqVA&uFp`4Fl[<.g6Mq'_`kE'5]j&,݀x]6/+2q.E'QYgƕ@oŀITKt!}e%Ar{~q8n>v_x>kjpfsa&&_z mrbo%XTM|]nLC Rcƹ= }j]{|3Ƕ[]=$;ϰ㲞|!Xnjm\Y9 Y@BΏ״͹g O>׶uT^PeTPERW=b Lhrv 4 ́v?,/S;L|?RP.,AN<+\؎L6$~њii :iI! 8inZViG)S[n%X&R~:@\ٴ"I-W Nz\2V863n1!r61l+ (/397bA e> ƨ׿  f@yHos;XoכKh9bTlΫwc?\/Y\TǒKK 䒅r6%7O2ॳWe\|:II5@zl+q"uhbT#tsMHȈ[R/^#)*9rIr'\r!CgYUć:q{>TϬ=iNTnz^ϑzo` K@Xn/D󔠄O>jj`-;i^޵Omk2ؖ]= {2c[oQ⸻@xIr">?PM,K?l}Rǡ]RǗX^׀N 7b?۶vp${E]lxDGPEU q8z3x/"Ҡ}qjIPb 82_spB&{7N ?8ʚ-,,%GFI40 .iY"\(x-M[;6΃ YZrn_.@6!*ZJ.#ī|h`49]?o@;AuNa.c ,.rR}x?NntA ƏMh\i_ aH)]\f{'kIV%pbje1VW"r.ԅ:T@WEyZIYz|GN,W~K(0x 뛌j) fkUOp/~3k!zЫ?9PC*{0dw6>yh3+:MpZ4ޫ{w{(Ǫ%)߫3يm tv hêlI |'Яb-ДadLb=>+CRFkеѠt<=#X%ɗ]AEP-,^ptDTe;J Wh=i'{7$0 &FBW. .}?'/bc$^J9>h=.u/h!$jgۓl݇Ͳo/wiۦ#b`mY8ѡg.i9 TX!H$O<.zN{"QM!`ݦS49yt]/Nǒ*K{4rb_7\ہӬltm 7ma\6{A0LЭ#0oryn6o8O(0$;-s|[35`B$}ty|&Vv <'t{jgԠp0XVWBYUK'[%9l? 4?H.1gvzK|qU?bTiN19*>O}9<^,0]z/u endstream endobj 24 0 obj <>/ExtGState<>/Font<>/ProcSet[/PDF/Text]/XObject<<>>>> endobj 25 0 obj <>stream xN0 y 0'ql_A*mHҐxz :HԱ/~-\\" c@b>z~;o=a1{Z*jJ zԘcv)$a}ФO`d K° D_0g^c/ k%c6}񼴞_[V$E{佤- V}P3Ҭ>Od(kR=n[ՋI>ĵ>k^c>X;տ/sZ!,[jcoxԮdլ_h gw꼽e _", endstream endobj 26 0 obj <> endobj 27 0 obj [28 0 R 29 0 R 30 0 R 31 0 R 32 0 R 33 0 R 34 0 R 35 0 R 36 0 R 37 0 R 38 0 R 39 0 R 40 0 R 41 0 R 42 0 R 43 0 R 44 0 R 45 0 R 46 0 R 47 0 R 48 0 R 49 0 R 50 0 R 51 0 R] endobj 28 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:28. 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Microbiol. 14, 576 588 \(2016\).\r:119)/F 4/H/N/Rect[373.222 510.855 378.218 498.444]/Subtype/Link/Type/Annot>> endobj 46 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:45. Feist, A. M. & Palsson, B. O. What do cells actually want? Genome Biol. 17\ , 110 \(2016\).\r:120)/F 4/H/N/Rect[474.96 466.855 480.224 454.444]/Subtype/Link/Type/Annot>> endobj 47 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:46. Oyetunde, T., Bao, F. S., Chen, J. W., Martin, H. G. \n& Tang, Y. J. Leveraging\ knowledge engineering and machine learning for microbial bio- manufacturing. Biotechnol. Adv. 36, 1308 1315 \(2018\).\r:121\ )/F 4/H/N/Rect[491.782 257.855 496.878 245.444]/Subtype/Link/Type/Annot>> endobj 48 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:47. Tran, L. M., Rizk, M. L. & Liao, J. C. Ensemble modeling of metabolic networks\ . Biophys. J. 95, 5606 5617 \(2008\).\r:122)/F 4/H/N/Rect[498.074 257.855 503.169 245.444]/Subtype/Link/Type/Annot>> endobj 49 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:48. Costello, Z. & Martin, H. G. A machine learning approach to predict metabolic \ pathway dynamics from time- series multiomics data. NPJ Syst. Biol. Appl. 4, 1 14 \(2018\).\r:123)/F 4/H/N/Rect[534.129 235.855 539.599 223.444]/Subtype/Link/Type/Annot>> endobj 50 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:49. Medlock, G. L. etal. Inferring metabolic mechanisms of interaction within a d\ efined gut microbiota. \nCell Syst. 7, 245 257.e7 \(2018\).\r:124)/F 4/H/N/Rect[523.428 213.855 528.475 201.444]/Subtype/Link/Type/Annot>> endobj 51 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:50. Heckmann, D. etal. Machine learning applied \nto enzyme turnover numbers revea\ ls protein structural correlates and improves metabolic models. Nat. 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Smith, A. L. etal. Navigating wastewater energy recovery strategies: a life c\ ycle comparison of anaerobic membrane bioreactor and conventional treatment systems with anaerobic digestion. Environ. Sci. Tec\ hnol. 48, 5972 5981 \(2014\).\r:128)/F 4/H/N/Rect[215.789 686.855 220.885 674.444]/Subtype/Link/Type/Annot>> endobj 100 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:53. Balakrishnan, M. etal. Novel pathways for fuels and lubricants from biomass o\ ptimized using life- cycle greenhouse gas assessment. Proc. Natl Acad. Sci. USA 112, 7645 7649 \(2015\).\r:129)/F 4/H/N/Rect[222.081 686.855 227.176 674.444]/Subtype/Link/Type/Annot>> endobj 101 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:19. Nielsen, P. H. etal. A conceptual ecosystem model \nof microbial communities \ in enhanced biological phosphorus removal plants. 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Segregation of biomass in cyclic anaerobic/aerobic granular sludge allows the enrichment of anaerobic ammonium oxidizing\ bacteria at low temperatures. Enviro:144)/F 4/H/N/Rect[412.69 180.855 417.761 168.444]/Subtype/Link/Type/Annot>> endobj 116 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:62. Laureni, M. etal. Biomass segregation between bio \nfilm and flocs improves th\ e control of nitrite- oxidizing bacteria in mainstream partial nitritation and anammox processes. Water Res. 154, 104 116 \(\ 2019\).\r:145)/F 4/H/N/Rect[514.656 147.855 519.742 135.444]/Subtype/Link/Type/Annot>> endobj 117 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:63. Scarborough, M. J., Lawson, C. E., Hamilton, J. J., Donohue, T. J. & Noguera, \ D. R. 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Methods 16, 167 170 \(2019\). \nThis study, together with referenc\ e 112, provides new techniques to trans:200)/F 4/H/N/Rect[272.25 81.8547 280.39 69.4439]/Subtype/Link/Type/Annot>> endobj 180 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:Box 1 | A design build test learn cycle to create synthetic microbiomes with desired\ functions\rWe present a generalized design build test learn cycle for creating synthetic microbiomes with desired functions, \ integrating both top- down and bottom-:201)/F 4/H/N/Rect[472.234 708.855 476.679 696.444]/Subtype/Link/Type/Annot>> endobj 181 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:Fig. 3 | Building self- assembled and synthetic microbiomes. a | A protoc\ ol for assembling synthetic microbiomes from multiple microbiome sources. Complex microbiomes can be taken apart into key functi\ onal members using automated microflu:202)/F 4/H/N/Rect[494.754 708.855 499.199 696.444]/Subtype/Link/Type/Annot>> endobj 182 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:113. Mulat, D. G. etal. Quantifying contribution of synthrophic acetate oxidation\ to methane production in thermophilic anaerobic reactors by membrane \ninlet mass spectrometry. Environ. Sci. Technol. 8, 2\ 505 2511 \(2014\).\r:204)/F 4/H/N/Rect[518.876 466.855 526.519 454.444]/Subtype/Link/Type/Annot>> endobj 183 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:Fig. 4 | Testing microbiome function. a | Isotopic tracers combined with \ the metaproteome could be used to measure microbiome metabolic flux by analysing isotopic labelling patterns of short peptides r\ ather than amino acids \(metabolome\). :205)/F 4/H/N/Rect[477.716 257.855 482.188 245.444]/Subtype/Link/Type/Annot>> endobj 184 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:114. Kehe, J. etal. Massively parallel screening of synthetic microbial communiti\ es. Proc. Natl Acad. Sci. USA 116, 12804 12809 \(2019\). \nThis study develops a high- throughput phenotypic screen using dr\ oplet- based microfluidics that \ncan an:206)/F 4/H/N/Rect[543.116 224.855 550.658 212.444]/Subtype/Link/Type/Annot>> endobj 185 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:82. Huber, R. etal. Robo- Lector - a novel platform for automated high- throughp\ ut cultivations in microtiter plates with high information content. Microb. Cell Fact. 8, 1 15 \(2009\).\r:207)/F 4/H/N/Rect[380.159 180.855 385.208 168.444]/Subtype/Link/Type/Annot>> endobj 186 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:83. Clark, C. etal. Characterization of TAP Ambr 250 disposable bioreactors, as \ a reliable scale- down model for biologics process development. Biotechnol. Prog. 33, 478 489 \(2017\).\r:208)/F 4/H/N/Rect[386.38 180.855 391.429 168.444]/Subtype/Link/Type/Annot>> endobj 187 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:Box 2 | A toolbox for measuring microbiome function\rMulti- omics integration\r\ The ability to assemble genomes from metagenomic data28 has enabled the genome- resolved analysis of individual transcriptom\ es63 and proteomes118 within d:209)/F 4/H/N/Rect[483.704 158.855 488.049 146.444]/Subtype/Link/Type/Annot>> endobj 188 0 obj <>/ExtGState<>/Font<>/ProcSet[/PDF/Text]/XObject<>>> endobj 189 0 obj <>stream x}r+G]?|D9u OLt[-iFx(M@v8#_\{g%@jG,TU*s_[r~Zŗ.ſ\|}uxN=ǮX,Z\J\\WK*Ey_h|u_,+&[ԳAsEj٨~e;R/7ϋmV.jh?[_C˟g,/e>k/kM0o|?che13:fllz@+5 >{V3=we.E)U^Z4y]Y엋zQ6TUrY,b^j}-۪2FwեKt|js1̮G(*ۈMt~nyh˥?Y盷Y.1k,[<mUu+Ko W-rvEozvqj4O.dgrվÅm߸U2_tntޭ.ٍwxWlAi^Nݾ5frc^E 9?B6mևnw?Cw0(`uk9LI& 6̵Iosц+ܷ"z;}&5h\{ՋM]V3)};]hkEʬS +l?ߙ'hWHB{bOHՌIR{MVjB~z^I\GOk[R}P_-ܝGv؏ZzPn6m/aQĕ̳2s2Wmگ k;A.o<~VGԩ'~x0GԠaQnd_A=5dpYDZ,aYZ^A sSipۗFM-U^uOhqNly~M<{3ʦ׃ܟHj.l&t EUyh%a³S|};%EkYͬudyհw،kLwC N=d ҷQk 9eG II(egyrΓd ~npaS~2Eyt5ImU7f(J܁osQlEѬ95vsPp!O=npWEԽ$i%r&BL{a0;[~IsXx.ZBԤ}g{Orn=zXwe< (Hj-JzZB{bdFz ; |OfkInu+ "s-VQBZz(~7avyY'Ew{RMmA孜lˢiDS6AI.qhTrrmiŢs|Ҥג@EuBj5ah2 cF܌hh35p/j՚J|7~J*wnu$gaЦW$OY+仹'Je|RSu狪kpf7XOW=mۑ"VS˺(I6 KonH hL9j)t-ZG'ZZbZ2j-A-R "7VQO4¦%11=i=ePZ*Mo:h:N0:A QZ*ն7ij$J}A| `n񱍠#j">w)*'rolXV^C*FOOd[rI^!fsAiPCo;9H-UCE1{[cRr8:9Ț|{@t)e!OZ,9m4$,Z%H6IZ-#|,ƸY;l7hhNj9Qу_-!swH,<">I3@FLᭁB(hŘL/=ֽbfy+H)n[ى,9B` 8 MVˀo34(Ƕ "xogDD>oLthgD2P@ eg`3po1MC6`_X@f|Jߑ(h!ER >=hC;,26 _35AQi8#(@QbQN=KRY?DW\I EJ=VRETY+J)#ht{1 EhcD8+R#͊u*ӝ1M i䘮)x$4 a$\Av)0!L\ zct@}( .3P:iApA.zfP^( NCm!<x*ƂQq6E%(ŋ1E*HU$c;V!v)oIvB(NjH}1 Ƌs>d{<*7Z| Qog $֨6c*Ptc~5NKjwj&,Mx2Fm8_RRQ`90wI'#O g nu|"DAdDׂ>c4¢px0xwmT4~\*J}h|l:ej.j A,Re^"nX*C`Ds &bg`hȞZ:߹X~y3( 3` a6b0:f!h1aآ*8@VV' 銔kI9ltlCBsm3s@ 9p>oqp搣 "Bm㥰p8R6&yAa»ĝJ3φ>W"Hr 5@k6rjDT̂7{ xSħէҙx5tJqPj!C}E4 ΋0ihNdȋ`|1ޯ@ZzKjJ~B<$J3~гq\V&Cᱶ;< \<;~>5w5e.֊]͆/2KNU Qg|FwϚ|@{z6وk>,u.NU_ZHȲpGr n5!L3jCšw\h\eLFt 8pz3.ĩ"xtI\3F+}ʦJDţcAgRu22fV)UØ)[K;BMn*PK)_"x`^0*%Uᪧc6$V:VVr7{ as%Wd) F,F I{ޙCd?3("6{e7Bv  h f& .&]tۑFDVZ1'H3yK@&Ai$#Df' ߆l}?e2>ae@cP*@Iy%30T\{3h6I*L3[%B;'(/|ܴZ va \NG|XW祷gƀznPV8 Jkʣ+I_}7R )[c[TZpw 0n#ݟ&nv=޸椅RFNi&F' CSywꕫ#%\\Qf2qHQOD|CT $J$GJUH^%'B Pkhz'JʋrP"f4Z|9 t2i^j&8:]s$ݑ(M`:@:9]F\!],_Q%~ |\+IW0Jd V1Kۖ5KyBqMm-Wu\/ld: AXzNhaHls=M@#U]ZNOgmn".yG9ٸU˲lD&:Dsu> W@;[VmX7.Z~l mAyZ;+, z)D8hP!(l]V8A$T>}'1JXQI` D8g;D@c_fa#/b;do}+Zҍ,υ,W/,TLܛ)ZxJZ`[Hþ_sӧa[8%PP_*dS"MTvnr0\F0ԶV0GmIoz6 rkӸU$ߌ nwіa zleDDX8[HRc'r` "x T XDp:Q${xΈN!j(]3taԚo4A-DŽ(\%Th1}2T <umƪf px=3 =Ytz4ZS,ǵYđƢ,@瞑Yf%XܑJoַC&_g 9L1iETnC>ӽox͌)PhQW͔T0zK `a( }syj{}bBaO/Qyѥ1@8ӘL4%I#Y&PEB}ʓ*c3+ U*jBLiVid6v.,ÌvCڎU3bNЮ5W`14̄& 'ئj3)[9݌w>MA*jy\֝7e v"6Ձq}jEi#C;)IxQ.U_NӫJ^]]S%IoAķ:)q;+H'ȩآ9Z@ә@3DÚ=D86U^UfV{1&vQ~]4sͨ0$}46ruHWՆ*A¢ JبjaWsPD35#y|PQo6&DBM twqJJ%d뇢n*eݟv@͂y||Jl%_-C]s%?y0H7s2Ʉ.]1ϧKp,H|IbOޤQX[T9OZ%0i&W$yo>?VBݴM[YԪߛ1Ɍr~.QW q=H}CX#bwPJRp*6?*җH!1A8/li"MSX  9ϬuSE|v,m5va.Î"cG^ B=O&- ]jfnFƊ|M . y!s8АƁ=yn}7.5`jM4`؄^Jkd5QݸԸ)kSȬ76>^aWZEbV[Ћ-HH'EUIhM܃H}'jg1nS&fVd驳i`_"|}3)TڠFU. F_& Yӛ9 i+bE@IWםpF`6wjeh ,I6>k 1(u~l4Og7QdVD&>%`K]FRmr Eܑ28Qr${/oB/qrA'+#i{ϧ:_oY(w&@S[Mt:"dsgu.p˫Gj4+1mD/ɜr֟kPzn~eyw׿6w~,+NF-{di`oJ&Ask45D}Vz0uwWuϣ |o lWköC[\5ԇE= BRS*q(0 7с b$g9\#qВ Zr:m\36;aN!PIP-_3%=(P#a $s\~HuO`b#gf":wמD'7rO6ylVUGbή}pٸ*ܧBݧ/ )B0˾m+ԀJe.C-wG}[ĖP:xB0#UО)Yht !A OvJ˜$ $.X2 PL/@^>n"0zA@a"@I]̴ ЂaukZ8zxl[vyq3|8]I bpf0\KcHR,%'\ x rKt~So.WRmqDӞ(#a+<,RLq~Jȗ}) ֡y\|ʮ)J]pG{C UTNՠc/-]?@N_Jv1’6r*n)*dpU^@̕YO A&G#} y ݎQΰo-dۡpA#,)كMoےO)(ɲĦ fI>f#fcE96yH;O dA[b 1.ֹjL8$6Έdikxԩ >PG=7igug=O>u3W4#;"D\/8ɬapMGP`9 V|45Ǥy,Buv󞼎0i;IGN^trIcHk'D+cfo. +=/{N 9s#5냊F~B́! 煢_hf4DYnb$ِĆv]fJ9bݨjy,@ϐ6vW^N-MAd,NJ33{ E\/XFB61Xmf4ƉQ&XT%j(ҕVMg*rr 5|TnM`fJ Qt qA S)z-m}iӴų4 /Iy ϐeI & ,cT%9٣ ܸ9 ]M5qllapuOl7zre\2GtOL.+@&_!a3877omʡ2R#xC{ظCe0AFRqHIg1#9Ӡ׬.kqiG.wˡ08д 9xgd;nZ0DrmR_u/u>&) z64ǕQ^ID™}adhh\NZ<'!)s d]'R J6o="*Hۤ -*#厀5 v.KAIY փTDliƠS`~7e2,wYi܈Y7;֔<,HORSP[ˈ],"6UhPX=x$}ALxV, 0@jix<̎t#NUx $q*bqPeW>bx`:eHD3H]`:vF@!娢oɝ@&< z=j$s' TEv޾d wRrUji;LepLg >1F`u? 4#cV|o. ?YK]/\Ϲ7Sz)5q#gUb/m/nQUʤpſ\|}ŷ*(DN",dMa o|*BoSݴ: Еx/fӂY:t0fCn3>3'U6+V78=#LG?+q4h:?L=tRPxJ2/*kWʰooJ°ܯzu8%P~3ܡf)BCLsxu!R8[ x\^S]->ߗ;~pD #^nO yuf꾐'5M_^]Pд2"췞8Ky_Y*)> endobj 191 0 obj [192 0 R 193 0 R 194 0 R 195 0 R 196 0 R 197 0 R 198 0 R 199 0 R 200 0 R 201 0 R] endobj 192 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:115. Truong, D. T., Tett, A., Pasolli, E., Huttenhower, C. & Segata, N. Microbial \ strain- level population structure and genetic diversity diversity from metagenomes. Nat. Methods 12, 626 638 \(2017\).\r:24\ 2)/F 4/H/N/Rect[262.502 257.855 269.89 245.444]/Subtype/Link/Type/Annot>> endobj 193 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:116. Mosbk, F. etal. Identification of syntrophic \nacetate- oxidizing bacteria \ in anaerobic digesters by combined protein- based stable isotope probing and metagenomics. ISME J. 10, 2405 2418 \(2016\).\r\ :243)/F 4/H/N/Rect[188.214 235.855 196.506 223.444]/Subtype/Link/Type/Annot>> endobj 194 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:118. Hawley, A. K., Brewer, H. M., Norbeck, A. D., \nPasa- Toli, L. & Hallam, S. J\ . Metaproteomics reveals differential modes of metabolic coupling among ubiquitous oxygen minimum zone microbes. \nProc. Natl A\ cad. Sci. USA 111, 11395 11400 \n\(20:244)/F 4/H/N/Rect[199.484 235.855 207.777 223.444]/Subtype/Link/Type/Annot>> endobj 195 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:119. Bowen, J. L., Babbin, A. R., Kearns, P. J. & Ward, B. B. Connecting the dots:\ linking nitrogen cycle gene expression to nitrogen fluxes in marine sediment mesocosms. Front. Microbiol. 5, 1 10 \(2014\).\ \r:245)/F 4/H/N/Rect[331.562 103.855 339.761 91.4439]/Subtype/Link/Type/Annot>> endobj 196 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:120. He, Z. etal. GeoChip: a comprehensive microarray \nfor investigating biogeoch\ emical, ecological and environmental processes. ISME J. 1, 67 77 \(2007\).\r:246)/F 4/H/N/Rect[280.289 92.8547 288.194 80.4439]/Subtype/Link/Type/Annot>> endobj 197 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:121. Hellerstein, M. K. Invivo measurement of fluxes through metabolic pathways: \ the missing link in functional genomics and pharmaceutical research. Annu. Rev. Nutr. 23, 379 402 \(2003\).\r:247)/F 4/H/N/Rect[434.126 224.855 441.769 212.444]/Subtype/Link/Type/Annot>> endobj 198 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:122. Sauer, U. Metabolic networks in motion: 13C- based flux analysis. Mol. Sys\ t. Biol. 2, 62 \(2006\).\r:248)/F 4/H/N/Rect[442.965 224.855 450.608 212.444]/Subtype/Link/Type/Annot>> endobj 199 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:123. Antoniewicz, M. R., Kelleher, J. K. & Stephanopoulos, G. Elementary metabolit\ e units \(EMU\): a novel framework for modeling isotopic distributions. Metab. Eng. 9, 68 86 \(2007\).\r:249)/F 4/H/N/Rect[482.574 125.855 490.224 113.444]/Subtype/Link/Type/Annot>> endobj 200 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:124. Gebreselassie, N. A. & Antoniewicz, M. R. \n13C- metabolic flux analysis of\ co- cultures: a novel approach. Metab. Eng. 31, 132 139 \(2015\).\r:250)/F 4/H/N/Rect[393.832 103.855 401.475 91.4439]/Subtype/Link/Type/Annot>> endobj 201 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:146. Zengler, K. etal. EcoFABs: advancing microbiome science through standardized\ fabricated ecosystems. Nat. Methods 16, 567 571 \(2019\). \nThis article describes the construction and use \nof standardize\ d fabricated ecosystems for the develop:203)/F 4/H/N/Rect[228.834 560.794 236.162 551.528]/Subtype/Link/Type/Annot>> endobj 202 0 obj <>/ExtGState<>/Font<>/ProcSet[/PDF/Text]/XObject<>>> endobj 203 0 obj <>stream x]rq#bXphBE#i$TȂOXEۙwR$ǖ"$b7oɓ ~Wo:wW^}'w]?|'pnnl:573#UCPLa0r847ZbS/J(LHi%~i8aďE Ľ6>eaZxy^d~g [ꝏ ȼ<2L`+VAx~]\MK xaպfܻ7٦&J TL{k\Z7(6xJt#q[,􀻪vN0f*O7h[ W( Pዷ0wڦR So? cVϛrϿxwz-_~Fޢ :TۭP$}4(fn.M^D(v`SI*Gd/`-i5+/'S |u\-ϼ,K3? 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Mosbk, F. etal. Identification of syntrophic \nacetate- oxidizing bacteria\ in anaerobic digesters by combined protein- based stable isotope probing and metagenomics. ISME J. 10, 2405 2418 \(2016\).\ \r:251)/F 4/H/N/Rect[331.6 697.855 339.761 685.444]/Subtype/Link/Type/Annot>> endobj 210 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:125. Ghosh, A. etal. A peptide- based method for \n13C metabolic flux analysis \ in microbial communities. PLOS Comput. Biol. 10, e1003827 \(2014\). \nThis study develops a novel method for calculating met\ abolic fluxes in microbial communities :252)/F 4/H/N/Rect[210.163 653.855 217.853 641.444]/Subtype/Link/Type/Annot>> endobj 211 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:126. Nielsen, J. It Is all about metabolic fluxes. J. Bacteriol. 185, 7031 703\ 5 \(2003\).\r:253)/F 4/H/N/Rect[304.755 598.855 312.349 586.444]/Subtype/Link/Type/Annot>> endobj 212 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:127. Bey, M., Azzouzi, S., Weitzel, M., Wiechert, W. \n& Nh, K. The design of Flu\ xML: a universal \nmodeling language for 13C metabolic flux analysis. Front. Microbiol. 10, 1022 \(2019\).\r:254)/F 4/H/N/Rect[296.341 532.855 303.861 520.444]/Subtype/Link/Type/Annot>> endobj 213 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:128. McGlynn, S. E., Chadwick, G. L., Kempes, C. P. & Orphan, V. J. Single cell ac\ tivity reveals direct electron transfer in methanotrophic consortia. Nature 526, 531 535 \(2015\).\r:255)/F 4/H/N/Rect[438.829 598.855 446.399 586.444]/Subtype/Link/Type/Annot>> endobj 214 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:129. Picioreanu, C., Prez, J. & van Loosdrecht, M. C. M. Impact of cell cluster s\ ize on apparent half- saturation coefficients for oxygen in nitrifying sludge and biofilms. 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Nielsen, J. L. & Nielsen, P. H. Advances in microscopy: microautoradiography \ of single cells. Methods Enzymol. 397, 237 256 \(2005\).\r:257)/F 4/H/N/Rect[293.947 246.855 302.065 234.444]/Subtype/Link/Type/Annot>> endobj 223 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:131. Huang, W. E. etal. Raman- FISH: combining stable- isotope Raman spectroscopy\ and fluorescence \ninsitu hybridization for the single cell analysis \nof identity and function. Environ. Microbiol. 9, 187\ 8 1889 \(2007\).\r:258)/F 4/H/N/Rect[194.024 235.855 201.717 223.444]/Subtype/Link/Type/Annot>> endobj 224 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:132. Dunham, S. J. B., Ellis, J. F., Li, B. & Sweedler, J. V. Mass spectrometry\ imaging of complex microbial communities. Acc. Chem. Res. 50, 96 104 \(2017\).\r:259)/F 4/H/N/Rect[307.228 235.855 314.921 223.444]/Subtype/Link/Type/Annot>> endobj 225 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:133. Hatzenpichler, R. etal. Visualizing insitu translational activity for iden\ tifying and sorting slow- growing archaeal"bacterial consortia. Proc. Natl Acad. \nSci. USA 113, E4069 E4078 \(2016\). \nThis\ study develops a high- throughput appr:260)/F 4/H/N/Rect[186.796 213.855 194.484 201.444]/Subtype/Link/Type/Annot>> endobj 226 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:Box 2 | A toolbox for measuring microbiome function\rMulti- omics integration\r\ The ability to assemble genomes from metagenomic data28 has enabled the genome- resolved analysis of individual transcripto\ mes63 and proteomes118 within :261)/F 4/H/N/Rect[214.521 213.855 218.958 201.444]/Subtype/Link/Type/Annot>> endobj 227 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:133. Hatzenpichler, R. etal. Visualizing insitu translational activity for ide\ ntifying and sorting slow- growing archaeal"bacterial consortia. Proc. Natl Acad. \nSci. USA 113, E4069 E4078 \(2016\). \nThi\ s study develops a high- throughput app:262)/F 4/H/N/Rect[171.085 114.855 178.696 102.444]/Subtype/Link/Type/Annot>> endobj 228 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:134. Okabe, S., Satoh, H. & Watanabe, Y. Analysis of microbial structure and funct\ ion of nitrifying biofilms. Methods Ecol. Evol. 337, 213 224 \(2001\).\r:263)/F 4/H/N/Rect[282.002 70.8547 289.618 58.4439]/Subtype/Link/Type/Annot>> endobj 229 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:95. Riglar, D. T. etal. Engineered bacteria can function \nin the mammalian gut l\ ong- term as live diagnostics \nof inflammation. Nat. Biotechnol. 35, 653 658 \(2017\).\r:264)/F 4/H/N/Rect[184.677 59.8547 190.243 47.4439]/Subtype/Link/Type/Annot>> endobj 230 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:13. Nielsen, J. & Keasling, J. D. Engineering cellular metabolism. Cell 164, \ 1185 1197 \(2016\). \nThis review highlights experiences, success stories and challenges associated with implementing the DBTL c\ ycle for metabolic engineering.\r:265)/F 4/H/N/Rect[545.449 59.8547 550.658 47.4439]/Subtype/Link/Type/Annot>> endobj 231 0 obj <>/ExtGState<>/Font<>/ProcSet[/PDF/Text]/Properties<>/Shading<>/XObject<>>> endobj 232 0 obj <>stream x\I8cBrn{Z#tk(j 23C`f%sj^qŮKxlLVѫǵ;f?_0mpnwWt7W=]p5mq޸6n5/WmW/ηZ aCٕڔ=|wWׯww~ WwDx޿8ջHn~yfq.~:GW|^tܼχ[74Lѵ_mmnyz?^0[.d7Ej?g] J}oabnmjy㴫)*v{^-f zշ#s ^J@c<%Z/W[WhmxLa[ׯ*?ꫛM?'\yEnҩУ}n="Zպ-vȽ~n6OnW\ 67gzy'}E_ ?~U5C죄m" gZ@c(Lꩭo>sYN6[M2<;Ya*;wSv?nx秐׍8EY#1.ֿZIЖ-$Tw%ZL&J~sIxHFrޣvkfW'ӷ6.#K][xkl+kʹ?y6?[~=^/pߙԜ-2| tGlm~U=by0D6zzq`kHoOAZ6\0^}  ̗O {K& ,{n^$89/5Iv ܠ3-{~O>,-H7ߍfw~O}p$ԏV8߷Y[_.F~C.&e$\ui5I( HcB?͌7m;1aו`r/t}wͪ_w%I^I}~c"xZ# /Ufxݰ DAI>v36{ga BǪrWfJ+,`Zn:@zB v  +t@;tGPQ)47}7߰Z)(y#uE=LFc1nhi"7n}֓#hZY3aM0j` Bo^*P4(6쥼 Z]D%ɋulPqGv~j;f|^k6fKqLy (gnmf^ªY? 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ISME J. 10, 2405 2418 \(2016\).\r:\ 218)/F 4/H/N/Rect[85.9064 399.022 93.106 389.756]/Subtype/Link/Type/Annot>> endobj 276 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:124. Gebreselassie, N. A. & Antoniewicz, M. R. \n13C- metabolic flux analysis of \ co- cultures: a novel approach. Metab. Eng. 31, 132 139 \(2015\).\r:219)/F 4/H/N/Rect[104.155 339.022 111.355 329.756]/Subtype/Link/Type/Annot>> endobj 277 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:125. Ghosh, A. etal. A peptide- based method for \n13C metabolic flux analysis i\ n microbial communities. PLOS Comput. Biol. 10, e1003827 \(2014\). \nThis study develops a novel method for calculating meta\ bolic fluxes in microbial communities u:220)/F 4/H/N/Rect[112.359 339.022 119.558 329.756]/Subtype/Link/Type/Annot>> endobj 278 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:168. Orphan, V. J., Orphan, V. J., House, C. H. & Hinrichs, K. Methane- consuming \ archaea revealed by directly coupled isotopic and phylogenetic analysis. Science 484, 484 488 \(2013\).\r:221)/F 4/H/N/Rect[195.325 243.022 202.439 233.756]/Subtype/Link/Type/Annot>> endobj 279 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:169. Kaltenpoth, M., Strupat, K. & Svatoa, A. Linking metabolite production to tax\ onomic identity in environmental samples by \(MA\)LDI- FISH. ISME J. 10, 527 531 \(2015\).\r:222)/F 4/H/N/Rect[203.414 243.022 210.527 233.756]/Subtype/Link/Type/Annot>> endobj 280 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:132. Dunham, S. J. B., Ellis, J. F., Li, B. & Sweedler, J. V. Mass spectrometry im\ aging of complex microbial communities. Acc. Chem. Res. 50, 96 104 \(2017\).\r:223)/F 4/H/N/Rect[261.264 223.022 268.536 213.756]/Subtype/Link/Type/Annot>> endobj 281 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:132. Dunham, S. J. B., Ellis, J. F., Li, B. & Sweedler, J. V. Mass spectrometry i\ maging of complex microbial communities. Acc. Chem. Res. 50, 96 104 \(2017\).\r:224)/F 4/H/N/Rect[218.371 143.022 225.485 133.756]/Subtype/Link/Type/Annot>> endobj 282 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:170. Nuez, J., Renslow, R., Cliff, J. B. & Anderton, C. R. NanoSIMS for biologica\ l applications: current practices and analyses. Biointerphases 13, 03B301 \(2018\).\r:225)/F 4/H/N/Rect[222.778 133.022 229.834 123.756]/Subtype/Link/Type/Annot>> endobj 283 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:132. Dunham, S. J. B., Ellis, J. F., Li, B. & Sweedler, J. V. Mass spectrometry \ imaging of complex microbial communities. Acc. Chem. Res. 50, 96 104 \(2017\).\r:226)/F 4/H/N/Rect[251.941 103.022 259.054 93.7555]/Subtype/Link/Type/Annot>> endobj 284 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:171. Northen, T. R. etal. Clathrate nanostructures for mass spectrometry. 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A. \n& Venturelli, O. S. Rapid m\ icrobial interaction network inference in microfluidic droplets. Preprint at bioRxiv https://doi.org/10.1101/521823 \(20\ 19\).\r:241)/F 4/H/N/Rect[535.943 103.022 543.315 93.7555]/Subtype/Link/Type/Annot>> endobj 299 0 obj <>/ExtGState<>/Font<>/ProcSet[/PDF/Text]/XObject<>>> endobj 300 0 obj <>stream x}[H^ڏi`?m e bKtWvόSbJܒHHUV6~/}hDTJ99o TJ"K_.x.V_mesח_eۘ%w}{a]/ozwr/XaVсJ#%_u1{Acri#m6 BJԲdD\RP .{sIģ— |)ϳ$FbMcA/P)FRZ>Oǻ>)9 ,d@  Z40M hih@Pĵc>@#%yTK}YKTANJ+%O!hN 1Q{EZJa9'W0j]$iꬲQk:AK  :X-UmHazmXv:->ɂ癋@_ 81 vʕYeCX IrA)aǽ8Pi I@c&Ip]rE$cE1h.|r?Kž yGnm/X9. ߷c,[QsVfNlB 2f!S"hr#h+[mJNSҏc!#eAT`wndK5 İ?z"(_Z!={TG<9!,Fׂ{ oCo =M60ӃZzrX-ۊ,,KՊz"{_Yst?.'$g[D;>^Y^ P@sGѮt&]l_g<+ҫr}tqX a:nU;(3AID3\d`L:rU:-+i!ϳ0 "hd$3>%Sqa&(Fz|ljex1D faGEADG{`l&/6TK7cBj4Q< HNqSt]fIi?9&Y2! 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Introducing THOR, a model microbiome for genetic dissect\ ion of community behavior. mBio 10, e02846 18 \(2019\).\r:280)/F 4/H/N/Rect[334.188 631.855 341.858 619.444]/Subtype/Link/Type/Annot>> endobj 307 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:146. Zengler, K. etal. EcoFABs: advancing microbiome science through standardize\ d fabricated ecosystems. Nat. Methods 16, 567 571 \(2019\). \nThis article describes the construction and use \nof standardiz\ ed fabricated ecosystems for the develo:281)/F 4/H/N/Rect[332.311 543.855 339.761 531.444]/Subtype/Link/Type/Annot>> endobj 308 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:146. Zengler, K. etal. EcoFABs: advancing microbiome science through standardiz\ ed fabricated ecosystems. Nat. Methods 16, 567 571 \(2019\). \nThis article describes the construction and use \nof standardi\ zed fabricated ecosystems for the devel:282)/F 4/H/N/Rect[396.841 686.855 404.536 674.444]/Subtype/Link/Type/Annot>> endobj 309 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:147. Zhalnina, K., Zengler, K., Newman, D. & Northen, T. R. Need for laboratory ec\ osystems to unravel the structures. mBio 9, 1 8 \(2018\).\r:283)/F 4/H/N/Rect[405.749 686.855 413.444 674.444]/Subtype/Link/Type/Annot>> endobj 310 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:Fig. 5 | Learning fundamental principles for microbiome engineering. 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Anaerobic microbial metabolism can proceed \ close to thermodynamic limits. Nature 415, 454 456 \(2002\).\r:291)/F 4/H/N/Rect[200.912 136.855 208.687 124.444]/Subtype/Link/Type/Annot>> endobj 330 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:153. Phelan, V. V., Liu, W. T., Pogliano, K. & Dorrestein, P. C. Microbial metabol\ ic exchange the chemotype- to-\nphenotype link. Nat. Chem. Biol. 8, 26 35 \(2012\).\r:292)/F 4/H/N/Rect[268.81 92.8547 276.42 80.4439]/Subtype/Link/Type/Annot>> endobj 331 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:154. Louca, S. etal. Function and functional redundancy \nin microbial systems. N\ at. Ecol. Evol. 2, 936 943 \(2018\).\r:293)/F 4/H/N/Rect[271.784 70.8547 279.646 58.4439]/Subtype/Link/Type/Annot>> endobj 332 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:155. Ladau, J. & Eloe- Fadrosh, E. A. 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HsM(/lk .T`'L>c!8IO4wr_ҏ\fWTfr:L 7""ܑÜjTQ+3Pd27/ OETQF&|*P߿{S?~{י7_}OB/bqX&D endstream endobj 343 0 obj <> endobj 344 0 obj [345 0 R 346 0 R 347 0 R 348 0 R 349 0 R 350 0 R 351 0 R 352 0 R] endobj 345 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:156. Thompson, J. A., Oliveira, R. A., Djukovic, A., \nUbeda, C. & Xavier, K. B. Ma\ nipulation of the quorum sensing signal AI-2 affects the antibiotic- treated gut microbiota. Cell Rep. 10, 1861 1871 \(2015\)\ .\r:303)/F 4/H/N/Rect[332.246 719.855 339.761 707.444]/Subtype/Link/Type/Annot>> endobj 346 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:157. Arkin, A. P. etal. KBase: the United States Department of Energy Systems Bio\ logy Knowledgebase. \nNat. Biotechnol. 36, 566 569 \(2018\).\r:304)/F 4/H/N/Rect[254.907 631.855 262.49 619.444]/Subtype/Link/Type/Annot>> endobj 347 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:158. Morrell, W. C. etal. The experiment data depot: \na web- based software tool \ for biological experimental data storage, sharing, and visualization. ACS Synth. Biol. 6, 2248 2259 \(2017\).\r:305)/F 4/H/N/Rect[263.666 631.855 271.25 619.444]/Subtype/Link/Type/Annot>> endobj 348 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:159. Solden, L. M. etal. Interspecies cross- feeding orchestrates carbon degradat\ ion in the rumen ecosystem. Nat. Microbiol. 3, 1274 1284 \(2018\).\r:306)/F 4/H/N/Rect[231.364 499.855 238.93 487.444]/Subtype/Link/Type/Annot>> endobj 349 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:160. Podolsky, I. A. etal. Harnessing nature s anaerobes \nfor biotechnology and b\ ioprocessing. Annu. Rev. Chem. Biomol. Eng. 10, 105 128 \(2019\).\r:307)/F 4/H/N/Rect[477.522 730.855 485.086 718.444]/Subtype/Link/Type/Annot>> endobj 350 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:161. Swift, C. L., Brown, J. L., Seppl, S. & O Malley, M. A. Co- cultivation of the\ anaerobic fungus Anaeromyces robustus with Methanobacterium bryantii enhances transcription of carbohydrate active enzymes. \n\ J. Ind. Microbiol. Biotechnol. http:308)/F 4/H/N/Rect[486.255 730.855 493.819 718.444]/Subtype/Link/Type/Annot>> endobj 351 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:162. Lee, K. S. etal. An automated raman- based platform for the sorting of live \ cells by functional properties. Nat. Microbiol. 4, 1035 1048 \(2019\).\r:309)/F 4/H/N/Rect[544.819 697.855 552.756 685.444]/Subtype/Link/Type/Annot>> endobj 352 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatRevMicro_255.indd:111. Ronda, C., Chen, S. P., Cabral, V., Yaung, S. J. & \nWang, H. H. Metagenomic\ engineering of the mammalian gut microbiome insitu. Nat. 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Lawson William R. Harcombe Roland Hatzenpichler Stephen R. Lindemann Frank E. Löffler Michelle A. O’Malley Héctor García Martín Brian F. Pfleger Lutgarde Raskin Ophelia S. Venturelli David G. Weissbrodt Daniel R. Noguera Katherine D. McMahon 10.1038/s41579-019-0255-9 http://dx.doi.org/10.1038/s41579-019-0255-9 journal Nature Reviews Microbiology © 2019, Springer Nature Limited 1740-1534 10.1038/s41579-019-0255-9 springer.com springerlink.com 2010-04-23 True 10.1038/s41579-019-0255-9 springer.com springerlink.com Springer 2019-11-07T15:50:33+05:30 2019-11-07T15:49:34+05:30 2019-11-07T15:50:33+05:30 True VoR uuid:3bd596db-fe1c-46f7-8116-9458ea37a6fb uuid:5fb6405d-6f1e-41d2-b4e9-02c8aa968cec default 1 converted uuid:915c3c62-ef4e-4b94-8909-22d32bc212da converted to PDF/A-2b pdfToolbox 2019-11-07T15:50:33+05:30 2 B Christopher E. Lawson http://orcid.org/0000-0002-3473-1640 William R. 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