NaCl 1.0 MgCl2·6H2O 0.2 CaCl2·2H2O 0.02 K2HPO4 1.5 (NH4)2SO4 2.0 NaHCO3 9.997 144, W381829 Alanine 0.5 Trace metal solutionb 5.0 mL 408, A0884 Asparagine 0.5 Ascorbic acid 0.005 371, 795437 40800, B4501 Biotin 0.005 Aspartic acid 0.5 166, A9256 290, M9625 Methionine 0.5 Cysteine 0.5 228, W326305 2575, C8731 Ca-Pantothenate 0.005 Nicotinic acid 0.005 80, N4126 1185, Pyridoxine·HCl 0.005 Prolinec 0.5 453, P0380 Thiamine 360, T4625 0.005 Serine 0.5 576, a Based on the unit prices for each CDM component, CDM is less expensive than the complex medium. All chemical prices were obtained from Sigma. For comparison, relevant components in the complex medium were given; 5 g/L polypeptone ($383/kg, 297110, BD Biosciences), 5 g/L yeast extract ($231/kg, 211929, BD Biosciences). Please note that the medium costs can be reduced by further metabolic engineering of M. so that the strain does not require these amino acids and vitamins; this work is in progress. succiniciproducens b The metal solution contains per liter: 5 mL HCl, 10 g FeSO4·7H2O, 2.25 g ZnSO4·7H2O, 1 g CuSO4·5H2O, 0.5 g MnSO4·5H2O, 0.23 g Na2B4O7·10H2O, 2 g CaCl2·2H2O, and 0.1 g (NH4)6Mo7O24. c The concentration of proline used in fed-batch fermentation was 2.46 g/L.
NaCl 1.0 MgCl2·6H2O 0.2 CaCl2·2H2O 0.02 K2HPO4 1.5 (NH4)2SO4 2.0 NaHCO3 9.997 Alanine 0.5 144, W381829 Trace metal solutionb 5.0 mL Asparagine 0.5 408, A0884 Ascorbic acid 0.005 371, 795437 Biotin 0.005 40800, B4501 Aspartic acid 0.5 166, A9256 Methionine 0.5 290, M9625 Cysteine 0.5 228, W326305 Ca-Pantothenate 0.005 2575, C8731 Nicotinic acid 0.005 80, N4126 Pyridoxine·HCl 0.005 1185, Prolinec 0.5 453, P0380 Thiamine 0.005 360, T4625 Serine 0.5 576, a Based on the unit prices for each CDM component, CDM is less expensive than the complex medium. All chemical prices were obtained from Sigma. For comparison, relevant components in the complex medium were given; 5 g/L polypeptone ($383/kg, 297110, BD Biosciences), 5 g/L yeast extract ($231/kg, 211929, BD Biosciences). Please note that the medium costs can be reduced by further metabolic engineering of M. succiniciproducens so that the strain does not require these amino acids and vitamins; this work is in progress. b The metal solution contains per liter: 5 mL HCl, 10 g FeSO4·7H2O, 2.25 g ZnSO4·7H2O, 1 g CuSO4·5H2O, 0.5 g MnSO4·5H2O, 0.23 g Na2B4O7·10H2O, 2 g CaCl2·2H2O, and 0.1 g (NH4)6Mo7O24. c The concentration of proline used in fed-batch fermentation was 2.46 g/L.
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