The composition of macromolecular cellular composition of mouse cell lines can be found
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1 Supplementary 2: Cell biomass composition The composition of macromolecular cellular composition of mouse cell lines can be found elsewhere. 1-5 Following table shows the percentage composition of important macromolecules which make up the cell. Calculated cellular composition Macromolecules %DCW DNA 1.9 RNA 5.8 Lipids 7.7 Carbohydrates 7.0 Proteins 70.6 Amino Acids Composition for proteins The amino acids content of the cell proteins were obtained from Bonarius et al. 6 Altamirano et al, 1 and Zupke et al. 7 Free amino acids in the cell were assumed to be negligible as it comprises only a small portion of the total cell components. Amino Acids MWt Average Contribution to protein g/mol Mole Fraction g/mol mmol/gdcw Ala Arg Asp Asn Cys Gln Glu Gly His Ile Leu Lys Met Phe Pro Ser Thr Trp
2 Tyr Val Monomers for DNA and RNA The nucleotide compositions were adopted from Gambhir et al. 8 and Sheikh et al. 3 The composition of monomers which make up DNA and RNA are listed below. Avg Mwt Average Composition Components Mole fraction g/mol g/mol (mol/mol) g/mol mmol/gdcw DNA DAMP DGMP DCMP DMMP RNA AMP GMP CMP UMP Lipid Composition The important components of lipids were obtained from literature. 9,10 The detailed composition of individual monomers were calculated and shown below. Lipid Composition Mwt wt% gm/mol mol/mol mmol/gdcw Cholesterol Phosphatidylcholine Phosphatidylethanolamine Phosphatidylinositol Phosphatidylserine Phosphatidylglycerol Diphosphatidylglycerol Sphingomyelin Av Mwt
3 Fatty Acid Composition Fatty acid composition was obtained from Bonarius et al. 6 and Sheikh et al. 3 The detailed composition is given below. Fatty Acid (w/w) % MW Contribution to avg FA g/mol g/mol mmol/g lipid mol/mol C14: C16: C16: C18: C18: C18: C20: C20: C20: C20: C22: Carbohydrate composition Glycogen was assumed to be the main carbohydrate content of the cell and its composition is given below. Carbohydrate Mwt wt% mmol/gdcw Glycogen Theoretical ATP requirement for cell biomass production Since biosynthesis of cell components require energy in the form of ATP, it is necessary to include ATP demand as well. The biosynthetic ATP demand was adopted from Xie and Wang. 11 Theoretical maximum ATP demand for cell biomass formation ATP Demand Percentage (mmol ATP/gDCW) Biosynthesis Cellular proteins Carbohydrates
4 Lipids RNA DNA Other important cell components: Apart from the main cellular components, there are others such as cofactors, vitamins and polyamines, the details of which for mouse cell lines were not available. However such values were adopted from other literature sources.
5 Overall Cellular Composition for Mouse: The overall average cell composition can be obtained from the calculation of individual macromolecular compositions and the detailed list of average cell composition is given below CELL COMPONENTS METABOLITES Composition (mmol/gdcw) Biomass Composition CELL COMPONENTS METABOLITES Composition (mmol/gdcw) PROTEINS ALA RNA CTP ARG UTP ASP LIPIDS CHOLESTEROL ASN PHOSPHATIDYLCHOLINE CYS PHOSPHATIDYLETHANOLAMINE GLN PHOSPHATIDYLINOSITOL GLU PHOSPHATIDYLSERINE GLY PHOSPHATIDYLGLYCEROL HIS DIPHOSPHATIDYLGLYCEROL ILE SPHINGOMYELIN LEU ENERGY ATP LYS CARBOHYDRATES GLYCOGEN MET H2O PHE COFACTORS NAD PRO NADP SER FAD THR NADH TRP NADPH TYR SUCCOA VAL ACCOA DNA DATP COA DGTP MTHF DCTP POLYAMINES PTRSC
6 DMTP SPRMD RNA AMP ANA GTP
7 References 1. C. Altamirano, A. Illanes, A. Casablancas, X. Gamez, J. J. Cairo and C. Godia, Biotechnol. Prog., 2001, 17, B. D. Follstad, R. R. Balcarcel, G. Stephanopoulos and D. I. C. Wang, Biotechnol. Bioeng., 1999, 63, K. Sheikh, J. Forster and L. K. Nielsen, Biotechnol. Prog., 2005, 21, L. Xie and D. I. C. Wang, Biotechnol. Bioeng., 1994, 43, L. Xie and D. I. C. Wang, Biotechnol. Bioeng., 1996, 52, H. P. J. Bonarius, V. Hatzimanikatis, K. P. H. Meesters, C. D. d. Gooijer, G. Schmid and J. Tramper, Biotechnol. Bioeng., 1996, 50, C. Zupke, A. J. Sinskey and G. Stephanopoulos, Appl. Microbiol. Biotechnol., 1995, 44, A. Gambhir, R. Korke, J. Lee, P.-C. Fu, A. F. Europa and W.-S. Hu, J. Biosci. Bioeng., 2003, 95, A. A. Spector and M. A. Yorek, J. Lipid Res., 1985, 26, Y. Nimura and I. Ishizuka, Biochemica et Biophysica Acta, 1981, 663, L. Xie and D. I. C. Wang, Biotechnol. Bioeng., 1996, 52,
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