Supplenentary text 2A diff from Ec_iAF1260_flux2.xml {Feist, 2007 #381} to Ec_iAF1260_noisozyme.xml used for initial FBA *** Ec_iAF1260_flux2.xml 2007-04-05 12:25:02.000000000 +0900 --- Ec_iAF1260_noisozyme.xml 2009-04-07 22:46:02.000000000 +0900 *************** *** 30794,30801 **** ! GENE_ASSOCIATION: ( b2097 or b1773 or b2925 ) ! PROTEIN_ASSOCIATION: ( FbaB ) or ( B1773 ) or ( FbaA ) SUBSYSTEM: S_GlycolysisGluconeogenesis PROTEIN_CLASS: 4.1.2.13 --- 30794,30801 ---- ! GENE_ASSOCIATION: b2925 ! PROTEIN_ASSOCIATION: FbaA SUBSYSTEM: S_GlycolysisGluconeogenesis PROTEIN_CLASS: 4.1.2.13 *************** *** 30821,30828 **** ! GENE_ASSOCIATION: ( b3925 or b4232 ) ! PROTEIN_ASSOCIATION: ( GlpX ) or ( Fbp ) SUBSYSTEM: S_GlycolysisGluconeogenesis PROTEIN_CLASS: 3.1.3.11 --- 30821,30828 ---- ! GENE_ASSOCIATION: b4232 ! PROTEIN_ASSOCIATION: Fbp SUBSYSTEM: S_GlycolysisGluconeogenesis PROTEIN_CLASS: 3.1.3.11 *************** *** 55225,55232 **** ! GENE_ASSOCIATION: ( b3916 or b1723 ) ! PROTEIN_ASSOCIATION: ( PfkA ) or ( PfkB ) SUBSYSTEM: S_GlycolysisGluconeogenesis PROTEIN_CLASS: 2.7.1.11 --- 55225,55232 ---- ! GENE_ASSOCIATION: b3916 ! PROTEIN_ASSOCIATION: PfkA SUBSYSTEM: S_GlycolysisGluconeogenesis PROTEIN_CLASS: 2.7.1.11 *************** *** 55693,55700 **** ! GENE_ASSOCIATION: ( b3612 or b4395 or b0755 ) ! PROTEIN_ASSOCIATION: ( YibO ) or ( GpmB ) or ( GpmA ) SUBSYSTEM: S_GlycolysisGluconeogenesis PROTEIN_CLASS: 5.4.2.1 --- 55693,55700 ---- ! GENE_ASSOCIATION: b0755 ! PROTEIN_ASSOCIATION: GpmA SUBSYSTEM: S_GlycolysisGluconeogenesis PROTEIN_CLASS: 5.4.2.1 *************** *** 60705,60712 **** ! GENE_ASSOCIATION: ( b1854 or b1676 ) ! PROTEIN_ASSOCIATION: ( Pyka ) or ( Pykf ) SUBSYSTEM: S_GlycolysisGluconeogenesis PROTEIN_CLASS: 2.7.1.40 --- 60705,60712 ---- ! GENE_ASSOCIATION: b1676 ! PROTEIN_ASSOCIATION: Pykf SUBSYSTEM: S_GlycolysisGluconeogenesis PROTEIN_CLASS: 2.7.1.40 *************** *** 61808,61815 **** ! GENE_ASSOCIATION: ( b3386 or b4301 ) ! PROTEIN_ASSOCIATION: ( Rpeec ) or ( SgcE ) SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 5.1.3.1 --- 61808,61815 ---- ! GENE_ASSOCIATION: b3386 ! PROTEIN_ASSOCIATION: Rpeec SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 5.1.3.1 *************** *** 61834,61841 **** ! GENE_ASSOCIATION: ( b2914 or b4090 ) ! PROTEIN_ASSOCIATION: ( RpiA ) or ( RpiB ) SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 5.3.1.6 --- 61834,61841 ---- ! GENE_ASSOCIATION: b2914 ! PROTEIN_ASSOCIATION: RpiA SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 5.3.1.6 *************** *** 63643,63650 **** ! GENE_ASSOCIATION: ( b2464 or b0008 ) ! PROTEIN_ASSOCIATION: ( TalA ) or ( TalB ) SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 2.2.1.2 --- 63643,63650 ---- ! GENE_ASSOCIATION: b0008 ! PROTEIN_ASSOCIATION: TalB SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 2.2.1.2 *************** *** 64824,64831 **** ! GENE_ASSOCIATION: ( b2935 or b2465 ) ! PROTEIN_ASSOCIATION: ( TktA ) or ( TktB ) SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 2.2.1.1 --- 64824,64831 ---- ! GENE_ASSOCIATION: b2935 ! PROTEIN_ASSOCIATION: TktA SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 2.2.1.1 *************** *** 64852,64859 **** ! GENE_ASSOCIATION: ( b2935 or b2465 ) ! PROTEIN_ASSOCIATION: ( TktA ) or ( TktB ) SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 2.2.1.1 --- 64852,64859 ---- ! GENE_ASSOCIATION: b2935 ! PROTEIN_ASSOCIATION: TktA SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 2.2.1.1 Supplenentary text 2B diff from Ec_iAF1260_noisozyme.xml to Ec_iAF1260_noisozyme_S17P.xml used for second FBA considering result from double knockout *** Ec_iAF1260_noisozyme.xml 2009-04-07 22:46:02.000000000 +0900 --- Ec_iAF1260_noisozyme_S17P.xml 2009-04-08 23:34:45.000000000 +0900 *************** *** 1437,1442 **** --- 1437,1443 ---- + *************** *** 55252,55257 **** --- 55253,55316 ---- + + + GENE_ASSOCIATION: b3916 + PROTEIN_ASSOCIATION: PfkA + SUBSYSTEM: S_Pentose_Phosphate_Pathway + PROTEIN_CLASS: 2.7.1.11 + + + + + + + + + + + + + FLUX_VALUE + + + + + + + + + + + + + + + GENE_ASSOCIATION: b2925 + PROTEIN_ASSOCIATION: FbaA + SUBSYSTEM: S_Pentose_Phosphate_Pathway + PROTEIN_CLASS: 4.1.2.13 + + + + + + + + + + + FLUX_VALUE + + + + + + + + + + GENE_ASSOCIATION: b3916 *************** *** 64824,64831 **** ! GENE_ASSOCIATION: b2935 ! PROTEIN_ASSOCIATION: TktA SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 2.2.1.1 --- 64883,64890 ---- ! GENE_ASSOCIATION: ( b2935 or b2465 ) ! PROTEIN_ASSOCIATION: ( TktA ) or ( TktB ) SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 2.2.1.1 *************** *** 64852,64859 **** ! GENE_ASSOCIATION: b2935 ! PROTEIN_ASSOCIATION: TktA SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 2.2.1.1 --- 64911,64918 ---- ! GENE_ASSOCIATION: ( b2935 or b2465 ) ! PROTEIN_ASSOCIATION: ( TktA ) or ( TktB ) SUBSYSTEM: S_Pentose_Phosphate_Pathway PROTEIN_CLASS: 2.2.1.1 Supplenentary text 2C RunFBAmodel1.m Script for initial FBA prediction using COBRA toolbox {Becker, 2007 #544}. clc % with alterarions concering discrepancy in single deletion % model used % without isozyme => 1 % with isozyme => 2 use_model = 1; % method used % linearMOMA => 1 % FBA => 2 use_method = 2; % airation condition % aerobic o2 > -18 => 1 % semi aerobic o2 > -5 => 2 % anaerobic => 3 % semi aerobic o2 > -8 => 4 % semi aerobic o2 > -12 => 5 % aerobic o2 > -8 => 6 % aerobic o2 > -12 => 7 air_cond = 1; % read model switch use_model case 1 disp('Ec_iAF1260_flux noisozyme...'); model = readCbModel('../Ec_iAF1260_noisozyme'); case 2 disp('Ec_iAF1260_flux2...'); model = readCbModel('../Ec_iAF1260_flux2'); end % air_cond switch air_cond case 1 disp('aerobic(o2 18)...'); model = changeRxnBounds(model,'EX_o2(e)' , -18, 'l'); case 2 disp('semi aerobic(o2 5)...'); model = changeRxnBounds(model,'EX_o2(e)' , -5, 'l'); model = changeRxnBounds(model, {'CADVtpp','DMSOR1','DMSOR2','FRD2','FRD3','HYD1pp','HYD2pp','HYD3pp',... 'NO2t2rpp','NTRIR2x','PFL','RNTR1c','RNTR2c','RNTR3c','RNTR4c','TMAOR1','TMAOR2'}, 100, 'u'); model = changeRxnBounds(model,'NO2t2rpp' , -100, 'l'); model = changeRxnBounds(model,'CYTBO3_4pp' , 0, 'b'); case 3 disp('anaerobic...'); model = changeRxnBounds(model,'EX_o2(e)' , 0, 'b'); model = changeRxnBounds(model, {'CADVtpp','DMSOR1','DMSOR2','FRD2','FRD3','HYD1pp','HYD2pp','HYD3pp',... 'NO2t2rpp','NTRIR2x','PFL','RNTR1c','RNTR2c','RNTR3c','RNTR4c','TMAOR1','TMAOR2'}, 100, 'u'); model = changeRxnBounds(model,'NO2t2rpp' , -100, 'l'); model = changeRxnBounds(model,'CYTBO3_4pp' , 0, 'b'); case 4 disp('semi aerobic(o2 8)...'); model = changeRxnBounds(model,'EX_o2(e)' , -8, 'l'); model = changeRxnBounds(model, {'CADVtpp','DMSOR1','DMSOR2','FRD2','FRD3','HYD1pp','HYD2pp','HYD3pp',... 'NO2t2rpp','NTRIR2x','PFL','RNTR1c','RNTR2c','RNTR3c','RNTR4c','TMAOR1','TMAOR2'}, 100, 'u'); model = changeRxnBounds(model,'NO2t2rpp' , -100, 'l'); model = changeRxnBounds(model,'CYTBO3_4pp' , 0, 'b'); case 5 disp('semi aerobic(o2 12)...'); model = changeRxnBounds(model,'EX_o2(e)' , -12, 'l'); model = changeRxnBounds(model, {'CADVtpp','DMSOR1','DMSOR2','FRD2','FRD3','HYD1pp','HYD2pp','HYD3pp',... 'NO2t2rpp','NTRIR2x','PFL','RNTR1c','RNTR2c','RNTR3c','RNTR4c','TMAOR1','TMAOR2'}, 100, 'u'); model = changeRxnBounds(model,'NO2t2rpp' , -100, 'l'); model = changeRxnBounds(model,'CYTBO3_4pp' , 0, 'b'); case 6 disp('aerobic(o2 8)...'); model = changeRxnBounds(model,'EX_o2(e)' , -8, 'l'); case 7 disp('aerobic(o2 12)...'); model = changeRxnBounds(model,'EX_o2(e)' , -12, 'l'); end % output file file = [ 'result_' num2str( use_model ) '_' num2str( air_cond ) '_' num2str( use_method ) '.txt' ]; fid = fopen( file,'w' ); % medium condition % Casamino Acid model = changeRxnBounds(model, {'EX_ala_L(e)','EX_arg_L(e)','EX_asp_L(e)','EX_cys_L(e)','EX_glu_L(e)',... 'EX_gly(e)','EX_his_L(e)','EX_ile_L(e)','EX_leu_L(e)','EX_lys_L(e)','EX_met_L(e)','EX_phe_L(e)','EX_pro_L(e)',... 'EX_ser_L(e)','EX_thr_L(e)','EX_trp_L(e)','EX_tyr_L(e)','EX_val_L(e)'},... [-0.21 -0.08 -0.22 0.00 -0.62 -0.12 -0.07 -0.18 -0.22 -0.23 -0.05 -0.12 -0.41 -0.11 -0.08 0.00 -0.01 -0.28],'l'); % carbon source CarbonSourceCond = {'noC','glc','mnl','fru','glyc','ac','succ','glu_L','glcur','glcn','xyl','mix'}; CarbonSourceName = {'EX_glc(e)','EX_mnl(e)','EX_fru(e)','EX_glyc(e)','EX_ac(e)',... 'EX_succ(e)','EX_glu_L(e)','EX_glcur(e)','EX_glcn(e)','EX_xyl_D(e)'}; CarbonSourceList = [0 0 0 0 0 0 0 0 0 0]; CarbonSourceList = [CarbonSourceList;-6 * eye(length(CarbonSourceList))]; CarbonSourceList = [CarbonSourceList;[0 0 0 -4 0 -4 0 0 0 0] ]; % additional glu_L(e) for i = 1 : length(CarbonSourceList) CarbonSourceList(i,7) = CarbonSourceList(i,7) + (-0.62); end % 2nd deletion list RxnDeletionList = {'-----','PGI','G6PDH2r','ICL','PPC','ME1','TALA','PYK'}; % 1st deletion list GeneDeletionList = {'-----','b0114','b0115','b2779','b2925','b2097','b4232','b1779','b2388','b0720',... 'b2029','b0755','b4395','b3612','b1136','b0116','b2463','b3403','b3916','b1723','b4025',... 'b2926','b3956','b1702','b1854','b1676','b3386','b2914','b4090','b1479','b2464','b0008',... 'b2935','b2465','b3919','b1852'}; % altlations to the model for mimic single gene deletion % for tpiA model = changeRxnBounds(model,'G3PD2' , -0.2, 'l'); %for TpiA; required for production of phospholipid but excess flux lead to growth of tpiA by exporting DHA via DHAP -> glyc3P -> glyc -> DHA model = changeRxnBounds(model,'F6PA' , 0, 'b'); %for TpiA; presence of this reaction is not confirmed in vivo and contradictory for the phenotypes of many mutants, like pgi-zwf, fbaA, pfkAB, model = changeRxnBounds(model,'EDD' , 0, 'b'); % for TpiA; known to be repressed except for gluconate and other carbon sources model = changeRxnBounds(model,'GLCDpp' , 0, 'b'); % for tpiA; this eraction is not active without addition of cofacter PQQ model = changeRxnBounds(model,'MGSA' , 0.25, 'u'); %for TpiA; known to operate in TpiA model = changeRxnBounds(model,'DRPA' , 0.25, 'u'); % for TpiA; R5P to nucleotides and degradation % for PDH; limit production of AcCoA model = changeRxnBounds(model,'POX' , 0.1, 'u'); % for from pyruvate to acetate model = changeRxnBounds(model,'ACALD' , 0.1, 'u'); % stop from acetoaldehyde model = changeRxnBounds(model,'ALDD2x' , 0.1, 'u'); % stop from acetoaldehyde via acetate model = changeRxnBounds(model,'CITL' , 0.1, 'u'); % stop from citrate via acetate model = changeRxnBounds(model,'GLYAT' , -0.1, 'l'); % stop threonine to glycone fprintf( fid, 'Gene1tGene2tSourcetmutnutYt'); for i = 1 : length(CarbonSourceName) fprintf( fid, '%st', CarbonSourceName{i}); end % additional flux shown fprintf( fid, 'RPItEX_o2(e)tPGItTALAtRPEtTKT1tPFKtPSKtICLtZwft'); fprintf( fid, 'n'); for i = 1 : length(RxnDeletionList) disp(RxnDeletionList{i}) if i == 1 modelx = model; else modelx = changeRxnBounds(model, RxnDeletionList{i}, 0, 'b'); % special alterations for 2ndary deletion if findRxnIDs(model,RxnDeletionList{i}) == findRxnIDs(model,'PPC') % PPC and PPCK modelx = changeRxnBounds(modelx, 'PPCK', 0, 'b'); elseif findRxnIDs(model,RxnDeletionList{i}) == findRxnIDs(model,'ME1') % ME1 and ME2 modelx = changeRxnBounds(modelx, 'ME2', 0, 'b'); end end for j = 1 : length(GeneDeletionList) disp(GeneDeletionList{j}) modelxx = modelx ; for k = 1 : length(CarbonSourceList) modelxxx = changeRxnBounds(modelxx,{'EX_glc(e)','EX_mnl(e)','EX_fru(e)','EX_glyc(e)','EX_ac(e)','EX_succ(e)',... 'EX_glu_L(e)','EX_glcur(e)','EX_glcn(e)','EX_xyl_D(e)'}, CarbonSourceList(k,:), 'l'); modelxxw = changeRxnBounds(model,{'EX_glc(e)','EX_mnl(e)','EX_fru(e)','EX_glyc(e)','EX_ac(e)','EX_succ(e)',... 'EX_glu_L(e)','EX_glcur(e)','EX_glcn(e)','EX_xyl_D(e)'}, CarbonSourceList(k,:), 'l'); % alterations in specific carbon source % {'noC','glc','mnl','fru','glyc','ac','succ','glu_L','glcur','glcn','xyl','mix'}; if k == 1 % Casamino acid only modelxxx = changeRxnBounds(modelxxx, { 'ICL', 'MALS' }, 100, 'u'); modelxxx = changeRxnBounds(modelxxx, 'TKT2', 0.1, 'u'); modelxxw = changeRxnBounds(modelxxw, { 'ICL', 'MALS' }, 100, 'u'); modelxxw = changeRxnBounds(modelxxw, 'TKT2', 0.1, 'u'); elseif k == 2 % glucose %glucose modelxxx = changeRxnBounds(modelxxx, 'PGI', -0.1, 'l'); %; limit reverseflux of PGI (thermodynamical constraint considering intracellular conc) modelxxw = changeRxnBounds(modelxxw, 'PGI', -0.1, 'l'); elseif k == 3 % mannitol modelxxx = changeRxnBounds(modelxxx, 'TKT2', 0.1, 'u'); %limit reverseflux of TKT2 (thermodynamical constraint considering intracellular conc) modelxxw = changeRxnBounds(modelxxw, 'TKT2', 0.1, 'u'); elseif k == 4 % flucose %frucose modelxxx = changeRxnBounds(modelxxx, 'TKT2', 0.1, 'u');; %limit reverseflux of TKT2 (thermodynamical constraint considering intracellular conc) modelxxw = changeRxnBounds(modelxxw, 'TKT2', 0.1, 'u'); elseif k == 5 % glycerol modelxxx = changeRxnBounds(modelxxx, { 'G3PD5', 'G3PD6', 'G3PD7', 'GLYK' }, 100, 'u'); modelxxx = changeRxnBounds(modelxxx, 'TKT2', 0.1, 'u'); modelxxw = changeRxnBounds(modelxxw, { 'G3PD5', 'G3PD6', 'G3PD7', 'GLYK' }, 100, 'u'); modelxxw = changeRxnBounds(modelxxw, 'TKT2', 0.1, 'u'); elseif k == 6 % acetate modelxxx = changeRxnBounds(modelxxx, 'ACS', 100, 'u'); modelxxx = changeRxnBounds(modelxxx, { 'ICL', 'MALS' }, 100, 'u'); modelxxx = changeRxnBounds(modelxxx, 'TKT2', 0.1, 'u'); modelxxw = changeRxnBounds(modelxxw, { 'ICL', 'MALS' }, 100, 'u'); modelxxw = changeRxnBounds(modelxxw, 'TKT2', 0.1, 'u'); modelxxw = changeRxnBounds(modelxxw, 'ACS', 100, 'u'); elseif k == 7 % succinate modelxxx = changeRxnBounds(modelxxx, { 'ICL', 'MALS' }, 100, 'u'); modelxxx = changeRxnBounds(modelxxx, 'TKT2', 0.1, 'u'); modelxxw = changeRxnBounds(modelxxw, { 'ICL', 'MALS' }, 100, 'u'); modelxxw = changeRxnBounds(modelxxw, 'TKT2', 0.1, 'u'); elseif k == 8 % glutamate modelxxx = changeRxnBounds(modelxxx, { 'ICL', 'MALS' }, 100, 'u'); modelxxx = changeRxnBounds(modelxxx, 'TKT2', 0.1, 'u'); modelxxw = changeRxnBounds(modelxxw, { 'ICL', 'MALS' }, 100, 'u'); modelxxw = changeRxnBounds(modelxxw, 'TKT2', 0.1, 'u'); elseif k == 9 % glucronate modelxxx = changeRxnBounds(modelxxx,'G6PDH2r' , 1, 'u'); modelxxw = changeRxnBounds(modelxxw,'G6PDH2r' , 1, 'u'); elseif k == 10 % gluconate modelxxx = changeRxnBounds(modelxxx,'EDD' , 100, 'u'); modelxxw = changeRxnBounds(modelxxw,'EDD' , 100, 'u'); modelxxx = changeRxnBounds(modelxxx,'G6PDH2r' , 1, 'u'); modelxxw = changeRxnBounds(modelxxw,'G6PDH2r' , 1, 'u'); elseif k == 11 % xylose modelxxx = changeRxnBounds(modelxxx, 'RPE', 0.1, 'u'); modelxxx = changeRxnBounds(modelxxx, 'TKT1', -0.1, 'l'); modelxxx = changeRxnBounds(modelxxx, 'TKT2', -0.1, 'l'); modelxxx = changeRxnBounds(modelxxx, { 'XYLI1', 'XYLK' , 'XYLabcpp' } , 100, 'u'); modelxxw = changeRxnBounds(modelxxw, 'RPE', 0.1, 'u'); modelxxw = changeRxnBounds(modelxxw, 'TKT1', -0.1, 'l'); modelxxw = changeRxnBounds(modelxxw, 'TKT2', -0.1, 'l'); modelxxw = changeRxnBounds(modelxxw, { 'XYLI1', 'XYLK' , 'XYLabcpp' } , 100, 'u'); elseif k == 12 % glycerol+Succinate modelxxx = changeRxnBounds(modelxxx, { 'G3PD5', 'G3PD6', 'G3PD7', 'GLYK' }, 100, 'u'); modelxxx = changeRxnBounds(modelxxx, { 'ICL', 'MALS' }, 100, 'u'); modelxxx = changeRxnBounds(modelxxx, 'TKT2', 0.1, 'u'); modelxxw = changeRxnBounds(modelxxw, { 'G3PD5', 'G3PD6', 'G3PD7', 'GLYK' }, 100, 'u'); modelxxw = changeRxnBounds(modelxxw, { 'ICL', 'MALS' }, 100, 'u'); modelxxw = changeRxnBounds(modelxxw, 'TKT2', 0.1, 'u'); end if i == 1 % reconfirm 2nd deletion modelxxx = modelxxx; else modelxxx = changeRxnBounds(modelxxx, RxnDeletionList{i}, 0, 'b'); end if GeneDeletionList{j} == '-----' modelxxx = modelxxx; else modelxxx = deleteModelGenes(modelxxx, GeneDeletionList{j}); end switch use_method case 1 solutionDel = linearMOMA(modelxxw,modelxxx,'max'); case 2 solutionDel = optimizeCbModel(modelxxx,'max',true,false); end mu = solutionDel.f; %mu fprintf( fid, '%st', RxnDeletionList{i}); fprintf( fid, '%st', GeneDeletionList{j}); fprintf( fid, '%st', CarbonSourceCond{k}); fprintf( fid, '%7.3ft', mu); if k == 1 | k == 12 fprintf( fid, '%7.3ft%7.3ft', 0, 0 ); else if length(solutionDel.x) == 0; fprintf( fid, 'nosolt'); else uptakeFlux = solutionDel.x(findRxnIDs(model,CarbonSourceName{k-1})); Y = mu/(-1 * uptakeFlux); fprintf( fid, '%7.3ft', uptakeFlux); fprintf( fid, '%7.3ft', Y); end end if length(solutionDel.x) == 0; fprintf( fid, 'nosolt'); else for l = 1 : length(CarbonSourceName) uptakeFlux = solutionDel.x(findRxnIDs(model,CarbonSourceName{l})); fprintf( fid, '%7.3ft', uptakeFlux); end fprintf( fid, '%7.3ft', solutionDel.x(findRxnIDs(modelxxx,'RPI'))); fprintf( fid, '%7.3ft', solutionDel.x(findRxnIDs(modelxxx,'EX_o2(e)'))); fprintf( fid, '%7.3ft', solutionDel.x(findRxnIDs(modelxxx,'PGI'))); fprintf( fid, '%7.3ft', solutionDel.x(findRxnIDs(modelxxx,'TALA'))); fprintf( fid, '%7.3ft', solutionDel.x(findRxnIDs(modelxxx,'RPE'))); fprintf( fid, '%7.3ft', solutionDel.x(findRxnIDs(modelxxx,'TKT1'))); fprintf( fid, '%7.3ft', solutionDel.x(findRxnIDs(modelxxx,'PFK'))); %fprintf( fid, '%7.3ft', solutionDel.x(findRxnIDs(modelxxx,'PSK'))); fprintf( fid, '%7.3ft', '-'); fprintf( fid, '%7.3ft', solutionDel.x(findRxnIDs(modelxxx,'ICL'))); fprintf( fid, '%7.3ft', solutionDel.x(findRxnIDs(modelxxx,'G6PDH2r'))); end modelxxw = model; modelxxx = modelxx; fprintf( fid, 'n'); end fprintf( fid, 'n'); modelxx = modelx; end modelx = model; fprintf( fid, 'n'); end Supplenentary text 2D diff from RunFBAmodel1.m to RunFBAmodel2.m Diference of Script for initial FBA and second FBA *** RunFBAmodel1.m 2009-05-01 19:23:14.000000000 +0900 --- RunFBAmodel2.m 2009-05-01 19:26:44.000000000 +0900 *************** *** 1,9 **** clc ! % with alterarions concering discrepancy in single deletion % model used % without isozyme => 1 % with isozyme => 2 ! use_model = 1; % method used % linearMOMA => 1 --- 1,11 ---- clc ! % with alterarions concering discrepancy in double deletion ! % model used % without isozyme => 1 % with isozyme => 2 ! % without isozyme,S17P => 3 ! use_model = 3; % method used % linearMOMA => 1 *************** *** 28,36 **** --- 30,42 ---- case 2 disp('Ec_iAF1260_flux2...'); model = readCbModel('../Ec_iAF1260_flux2'); + case 3 + disp('Ec_iAF1260_noisozyme_S17P...'); + model = readCbModel('../Ec_iAF1260_noisozyme_S17P'); end + % air_cond switch air_cond case 1 *************** *** 70,76 **** case 7 disp('aerobic(o2 12)...'); model = changeRxnBounds(model,'EX_o2(e)' , -12, 'l'); - end % output file --- 76,81 ---- *************** *** 79,85 **** % medium condition ! % Casamino Acid model = changeRxnBounds(model, {'EX_ala_L(e)','EX_arg_L(e)','EX_asp_L(e)','EX_cys_L(e)','EX_glu_L(e)',... 'EX_gly(e)','EX_his_L(e)','EX_ile_L(e)','EX_leu_L(e)','EX_lys_L(e)','EX_met_L(e)','EX_phe_L(e)','EX_pro_L(e)',... 'EX_ser_L(e)','EX_thr_L(e)','EX_trp_L(e)','EX_tyr_L(e)','EX_val_L(e)'},... --- 84,90 ---- % medium condition ! % Casamino Acid model = changeRxnBounds(model, {'EX_ala_L(e)','EX_arg_L(e)','EX_asp_L(e)','EX_cys_L(e)','EX_glu_L(e)',... 'EX_gly(e)','EX_his_L(e)','EX_ile_L(e)','EX_leu_L(e)','EX_lys_L(e)','EX_met_L(e)','EX_phe_L(e)','EX_pro_L(e)',... 'EX_ser_L(e)','EX_thr_L(e)','EX_trp_L(e)','EX_tyr_L(e)','EX_val_L(e)'},... *************** *** 99,142 **** end % 2nd deletion list RxnDeletionList = {'-----','PGI','G6PDH2r','ICL','PPC','ME1','TALA','PYK'}; % 1st deletion list GeneDeletionList = {'-----','b0114','b0115','b2779','b2925','b2097','b4232','b1779','b2388','b0720',... 'b2029','b0755','b4395','b3612','b1136','b0116','b2463','b3403','b3916','b1723','b4025',... 'b2926','b3956','b1702','b1854','b1676','b3386','b2914','b4090','b1479','b2464','b0008',... 'b2935','b2465','b3919','b1852'}; ! % altlations to the model for mimic single gene deletion ! ! % for tpiA ! model = changeRxnBounds(model,'G3PD2' , -0.2, 'l'); %for TpiA; required for production of phospholipid but excess flux lead to growth of tpiA by exporting DHA via DHAP -> glyc3P -> glyc -> DHA ! model = changeRxnBounds(model,'F6PA' , 0, 'b'); %for TpiA; presence of this reaction is not confirmed in vivo and contradictory for the phenotypes of many mutants, like pgi-zwf, fbaA, pfkAB, ! model = changeRxnBounds(model,'EDD' , 0, 'b'); % for TpiA; known to be repressed except for gluconate and other carbon sources ! model = changeRxnBounds(model,'GLCDpp' , 0, 'b'); % for tpiA; this eraction is not active without addition of cofacter PQQ ! model = changeRxnBounds(model,'MGSA' , 0.25, 'u'); %for TpiA; known to operate in TpiA ! model = changeRxnBounds(model,'DRPA' , 0.25, 'u'); % for TpiA; R5P to nucleotides and degradation ! % for PDH; limit production of AcCoA model = changeRxnBounds(model,'POX' , 0.1, 'u'); % for from pyruvate to acetate model = changeRxnBounds(model,'ACALD' , 0.1, 'u'); % stop from acetoaldehyde model = changeRxnBounds(model,'ALDD2x' , 0.1, 'u'); % stop from acetoaldehyde via acetate model = changeRxnBounds(model,'CITL' , 0.1, 'u'); % stop from citrate via acetate model = changeRxnBounds(model,'GLYAT' , -0.1, 'l'); % stop threonine to glycone ! fprintf( fid, 'Gene1\tGene2\tSource\tmu\tnu\tY\t'); for i = 1 : length(CarbonSourceName) fprintf( fid, '%s\t', CarbonSourceName{i}); end ! % additional flux shown fprintf( fid, 'RPI\tEX_o2(e)\tPGI\tTALA\tRPE\tTKT1\tPFK\tPSK\tICL\tZwf\t'); - fprintf( fid, '\n'); for i = 1 : length(RxnDeletionList) --- 104,159 ---- end % 2nd deletion list + %RxnDeletionList = {'ICL'}; RxnDeletionList = {'-----','PGI','G6PDH2r','ICL','PPC','ME1','TALA','PYK'}; % 1st deletion list GeneDeletionList = {'-----','b0114','b0115','b2779','b2925','b2097','b4232','b1779','b2388','b0720',... 'b2029','b0755','b4395','b3612','b1136','b0116','b2463','b3403','b3916','b1723','b4025',... 'b2926','b3956','b1702','b1854','b1676','b3386','b2914','b4090','b1479','b2464','b0008',... 'b2935','b2465','b3919','b1852'}; + %GeneDeletionList = {'-----','b4232','b3919','b0115'}; + %GeneDeletionList = {'-----'}; + % altlations to the model + % for TpiA + model = changeRxnBounds(model,'G3PD2' , -0.2, 'l'); %for TpiA + model = changeRxnBounds(model,'F6PA' , 0, 'b'); %for TpiA, pgi-zwf + model = changeRxnBounds(model,'EDD' , 0, 'b'); % except for gluconate + model = changeRxnBounds(model,'GLCDpp' , 0, 'b'); % to glcn 5dglcd and export ! model = changeRxnBounds(model,'MGSA' , 0, 'b'); % Activate only on TpiA mutant ! model = changeRxnBounds(model,'DRPA' , 0, 'b'); % for TpiA ! % for PDH model = changeRxnBounds(model,'POX' , 0.1, 'u'); % for from pyruvate to acetate model = changeRxnBounds(model,'ACALD' , 0.1, 'u'); % stop from acetoaldehyde model = changeRxnBounds(model,'ALDD2x' , 0.1, 'u'); % stop from acetoaldehyde via acetate model = changeRxnBounds(model,'CITL' , 0.1, 'u'); % stop from citrate via acetate model = changeRxnBounds(model,'GLYAT' , -0.1, 'l'); % stop threonine to glycone ! % for PckA-ME; stop TCA cycle to glycolysis ! model = changeRxnBounds(model,'THRD', 0, 'b'); % Thr to methylglyoxal ! model = changeRxnBounds(model,'THRAi', 0, 'b'); % Thr to methylglyoxal via gly ! model = changeRxnBounds(model,'GLXCL', 0, 'b'); % glyoxylate to 3PG ! model = changeRxnBounds(model,'OAADC', 0, 'b'); % oaa -> pyruvate + CO2 ! model = changeRxnBounds(model,'MCITS', 0, 'b'); % oaa to pyruvate by 2-methylisocitrate cycle ! ! % for rpiA; block exit from ru5p -> ara5p -> lipidA etc -> export ! model = changeRxnBounds(model, 'EX_kdo2lipid4(e)', 0, 'b'); ! model = changeRxnBounds(model, 'EX_lipa_cold(e)', 0, 'b'); ! model = changeRxnBounds(model, 'EX_lipa(e)', 0, 'b'); ! model = changeRxnBounds(model, 'EX_enlipa(e)', 0, 'b'); fprintf( fid, 'Gene1\tGene2\tSource\tmu\tnu\tY\t'); for i = 1 : length(CarbonSourceName) fprintf( fid, '%s\t', CarbonSourceName{i}); end ! % additional flux 0401 fprintf( fid, 'RPI\tEX_o2(e)\tPGI\tTALA\tRPE\tTKT1\tPFK\tPSK\tICL\tZwf\t'); fprintf( fid, '\n'); for i = 1 : length(RxnDeletionList) *************** *** 146,151 **** --- 163,169 ---- modelx = model; else modelx = changeRxnBounds(model, RxnDeletionList{i}, 0, 'b'); + % special alterations for 2ndary deletion if findRxnIDs(model,RxnDeletionList{i}) == findRxnIDs(model,'PPC') % PPC and PPCK *************** *** 153,158 **** --- 171,179 ---- elseif findRxnIDs(model,RxnDeletionList{i}) == findRxnIDs(model,'ME1') % ME1 and ME2 modelx = changeRxnBounds(modelx, 'ME2', 0, 'b'); + elseif findRxnIDs(model,RxnDeletionList{i}) == findRxnIDs(model,'TALA') + % Activate PSK on TALA deletion + modelx = changeRxnBounds(modelx, 'PSK', 100, 'u'); end end *************** *** 160,166 **** disp(GeneDeletionList{j}) modelxx = modelx ; ! for k = 1 : length(CarbonSourceList) modelxxx = changeRxnBounds(modelxx,{'EX_glc(e)','EX_mnl(e)','EX_fru(e)','EX_glyc(e)','EX_ac(e)','EX_succ(e)',... 'EX_glu_L(e)','EX_glcur(e)','EX_glcn(e)','EX_xyl_D(e)'}, CarbonSourceList(k,:), 'l'); --- 181,187 ---- disp(GeneDeletionList{j}) modelxx = modelx ; ! for k = 1 : length(CarbonSourceList) modelxxx = changeRxnBounds(modelxx,{'EX_glc(e)','EX_mnl(e)','EX_fru(e)','EX_glyc(e)','EX_ac(e)','EX_succ(e)',... 'EX_glu_L(e)','EX_glcur(e)','EX_glcn(e)','EX_xyl_D(e)'}, CarbonSourceList(k,:), 'l'); *************** *** 179,185 **** %glucose modelxxx = changeRxnBounds(modelxxx, 'PGI', -0.1, 'l'); %; limit reverseflux of PGI (thermodynamical constraint considering intracellular conc) modelxxw = changeRxnBounds(modelxxw, 'PGI', -0.1, 'l'); ! elseif k == 3 % mannitol modelxxx = changeRxnBounds(modelxxx, 'TKT2', 0.1, 'u'); %limit reverseflux of TKT2 (thermodynamical constraint considering intracellular conc) modelxxw = changeRxnBounds(modelxxw, 'TKT2', 0.1, 'u'); elseif k == 4 % flucose --- 200,207 ---- %glucose modelxxx = changeRxnBounds(modelxxx, 'PGI', -0.1, 'l'); %; limit reverseflux of PGI (thermodynamical constraint considering intracellular conc) modelxxw = changeRxnBounds(modelxxw, 'PGI', -0.1, 'l'); ! elseif k == 3 ! % mannitol modelxxx = changeRxnBounds(modelxxx, 'TKT2', 0.1, 'u'); %limit reverseflux of TKT2 (thermodynamical constraint considering intracellular conc) modelxxw = changeRxnBounds(modelxxw, 'TKT2', 0.1, 'u'); elseif k == 4 % flucose *************** *** 239,250 **** else modelxxx = changeRxnBounds(modelxxx, RxnDeletionList{i}, 0, 'b'); end ! ! if GeneDeletionList{j} == '-----' ! modelxxx = modelxxx; ! else ! modelxxx = deleteModelGenes(modelxxx, GeneDeletionList{j}); ! end switch use_method case 1 --- 261,282 ---- else modelxxx = changeRxnBounds(modelxxx, RxnDeletionList{i}, 0, 'b'); end ! ! if GeneDeletionList{j} == '-----' ! modelxxx = modelxxx; ! ! elseif GeneDeletionList{j} == 'b2029' % gnd ! modelxxx = deleteModelGenes(modelxxx, GeneDeletionList{j}); ! modelxxx = changeRxnBounds(modelxxx, 'EDD', 100, 'u'); ! elseif GeneDeletionList{j} == 'b0008' % talB ! modelxxx = deleteModelGenes(modelxxx, GeneDeletionList{j}); ! modelxxx = changeRxnBounds(modelxxx, 'PSK', 100, 'u'); ! elseif GeneDeletionList{j} == 'b3919' % tpiA ! modelxxx = deleteModelGenes(modelxxx, GeneDeletionList{j}); ! modelxxx = changeRxnBounds(modelxxx,'MGSA' , 0.5, 'u'); % Activate only on TpiA mutant ! else ! modelxxx = deleteModelGenes(modelxxx, GeneDeletionList{j}); ! end switch use_method case 1 *************** *** 290,297 **** fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'RPE'))); fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'TKT1'))); fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'PFK'))); ! %fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'PSK'))); ! fprintf( fid, '%7.3f\t', '-'); fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'ICL'))); fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'G6PDH2r'))); end --- 322,328 ---- fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'RPE'))); fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'TKT1'))); fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'PFK'))); ! fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'PSK'))); fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'ICL'))); fprintf( fid, '%7.3f\t', solutionDel.x(findRxnIDs(modelxxx,'G6PDH2r'))); end