Metagrowth: a database of evidences and hypotheses for the study of culture conditions of obligate parasitic bacteria

Total number of physical conditions and nutriments: 83

Hypothesis / Physical conditions or nutriments	Evidence type	Evidences / Treponema pallidum
(T=37C)	E	E0208 T. pallidum was incubated with and without cells in cell culture medium (pH 7.3), under deoxygenated conditions. Five to ten percent of the treponemes remained motile in cell-treponeme systems of co-incubation. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Continuous passage of the T. pallidum was not achieved in vitro [1]. A nonpathogenic treponeme were maintained and passaged in thioglycolate medium with 10% heat-inactivated normal rabbit serum at 37 degree C [2].
(pH=7.3)	E	E0208 T. pallidum was incubated with and without cells in cell culture medium (pH 7.3), under deoxygenated conditions. Five to ten percent of the treponemes remained motile in cell-treponeme systems of co-incubation. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Continuous passage of the T. pallidum was not achieved in vitro [1]. A nonpathogenic treponeme were maintained and passaged in thioglycolate medium with 10% heat-inactivated normal rabbit serum at 37 degree C [2].
(O2=3%)	E	E0201 T. pallidum is microaerophilic, which is sensitive but require oxygen (O2). The addition of sodium metabisulfite to a Eagle's minimal essential medium with rabbit serum was found to have an effect similar to that of dithiothreitol in the survival of T. pallidum under 3% O2. Analyses revealed that 50% motility was retained longest at O2 concentrations of 1-5%, whether or not dithiothreitol or sodium metabisulfite were present. Concentrations of O2 of 3-10% were optimal for nucleic acid synthesis. Sodium metabisulfite was less effective than dithiothreitol in stimulating nucleic acid synthesis. During incubation under 3% O2, motility was maintained at > 50% for 15 days. The optimal retention of motility and nucleic acid synthesis under microaerobic conditions in the absence of reducing compounds provides evidence that T. pallidum is a microaerophilic organism [1].
(O2=3.0%)	E	E0210 The effects of the concentrations of dithiothreitol, molecular oxygen, and antioxidants on the in vitro replication of T. pallidum were studied [1]. The optimal dithiothreitol concentration was 0.65-1.62 mM. The optimum oxygen concentration was 3.0%. The optimum concentrations for the water-soluble antioxidants cobalt chloride, cocarboxylase (e.g. thiamin-PP), mannitol, and histidine were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication. Histidine and mannitol were the most critical. Catalase and superoxide dismutase were investigated for their ability to promote the growth in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. The medium supported the replication of T. pallidum at oxygen concentrations of 5-7%. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth in this medium.
(O2=5.0-7.0%)	E	E0210 The effects of the concentrations of dithiothreitol, molecular oxygen, and antioxidants on the in vitro replication of T. pallidum were studied [1]. The optimal dithiothreitol concentration was 0.65-1.62 mM. The optimum oxygen concentration was 3.0%. The optimum concentrations for the water-soluble antioxidants cobalt chloride, cocarboxylase (e.g. thiamin-PP), mannitol, and histidine were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication. Histidine and mannitol were the most critical. Catalase and superoxide dismutase were investigated for their ability to promote the growth in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. The medium supported the replication of T. pallidum at oxygen concentrations of 5-7%. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth in this medium.
(CO2=x%)	E	E0208 T. pallidum was incubated with and without cells in cell culture medium (pH 7.3), under deoxygenated conditions. Five to ten percent of the treponemes remained motile in cell-treponeme systems of co-incubation. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Continuous passage of the T. pallidum was not achieved in vitro [1]. A nonpathogenic treponeme were maintained and passaged in thioglycolate medium with 10% heat-inactivated normal rabbit serum at 37 degree C [2].
(H2=x%)	E	E0208 T. pallidum was incubated with and without cells in cell culture medium (pH 7.3), under deoxygenated conditions. Five to ten percent of the treponemes remained motile in cell-treponeme systems of co-incubation. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Continuous passage of the T. pallidum was not achieved in vitro [1]. A nonpathogenic treponeme were maintained and passaged in thioglycolate medium with 10% heat-inactivated normal rabbit serum at 37 degree C [2].
Sugar C11477	T	E0216 Presence of transporters for sugar (TransportDB [1]), which include ribose, galactose, malate, succinate, glucose, glycerol-P [2].
D-Glucose C00031	STU	E0202 Glucose is incorporated only in pentose of nucleic acids, aspartate in proteins, glycerol of phosphatidylcholin and phosphatidylglycerol [1,2]. Exogenous choline was incorporated in phosphatidylcholin [2]. E0206 Predicted chemotaxis specificity includes aspartate, glutamate, histidine, glucose, and ribose [1]. E0216 Presence of transporters for sugar (TransportDB [1]), which include ribose, galactose, malate, succinate, glucose, glycerol-P [2].
Glucose C00293	S	E0241 Glycolytic pathway is present, in which two steps are catalyzed by unusual pyrophosphate-dependent enzymes (according to KEGG [1]).
D-Galactose C00124	ST	E0216 Presence of transporters for sugar (TransportDB [1]), which include ribose, galactose, malate, succinate, glucose, glycerol-P [2]. E0256 Galactose may enter in the glycolysis (according to KEGG [1]).
D-Galactoside C02023	T	E0220 Presence of transporters for galactosides (TransportDB [1]).
D-Mannose C00159	S	E0242 Fructose and mannose can enter in the glycolysis (according to KEGG [1]).
Mannitol C00392	E	E0210 The effects of the concentrations of dithiothreitol, molecular oxygen, and antioxidants on the in vitro replication of T. pallidum were studied [1]. The optimal dithiothreitol concentration was 0.65-1.62 mM. The optimum oxygen concentration was 3.0%. The optimum concentrations for the water-soluble antioxidants cobalt chloride, cocarboxylase (e.g. thiamin-PP), mannitol, and histidine were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication. Histidine and mannitol were the most critical. Catalase and superoxide dismutase were investigated for their ability to promote the growth in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. The medium supported the replication of T. pallidum at oxygen concentrations of 5-7%. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth in this medium.
D-Fructose C00095	S	E0242 Fructose and mannose can enter in the glycolysis (according to KEGG [1]).
Pyruvate C00022	E	E0208 T. pallidum was incubated with and without cells in cell culture medium (pH 7.3), under deoxygenated conditions. Five to ten percent of the treponemes remained motile in cell-treponeme systems of co-incubation. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Continuous passage of the T. pallidum was not achieved in vitro [1]. A nonpathogenic treponeme were maintained and passaged in thioglycolate medium with 10% heat-inactivated normal rabbit serum at 37 degree C [2].
Succinate C00042	T	E0216 Presence of transporters for sugar (TransportDB [1]), which include ribose, galactose, malate, succinate, glucose, glycerol-P [2].
(S)-Malate C00149	T	E0216 Presence of transporters for sugar (TransportDB [1]), which include ribose, galactose, malate, succinate, glucose, glycerol-P [2].
Glycerol 1-phosphate C00623	T	E0216 Presence of transporters for sugar (TransportDB [1]), which include ribose, galactose, malate, succinate, glucose, glycerol-P [2].
Glycerol 3-phosphate C00093	TU	E0204 There is no inorganic phosphate uptake system. Glycerol-3-phosphate through the multiple sugar transporter may be responsible for the source of phosphate [1]. E0216 Presence of transporters for sugar (TransportDB [1]), which include ribose, galactose, malate, succinate, glucose, glycerol-P [2].
Amino acid C00045	DGT	E0221 Presence of transporters for amino acid (TransportDB [1]). E0246 Except for a few interconversion enzymes, there are no biosynthetic pathways for different amino acids (according to KEGG [1]). We have to add all the 20 amino acids.
Peptide C00012	T	E0218 Presence of transporters for peptide (TransportDB [1]).
L-Alanine C00041	GT	E0225 Presence of transporters for sodium/alanine and alanine (TransportDB [1]). According to KEGG [2], TP0414 is D-alanine and glycine permease (dagA).
L-Aspartate C00049	GU	E0206 Predicted chemotaxis specificity includes aspartate, glutamate, histidine, glucose, and ribose [1].
Glycine C00037	GT	E0225 Presence of transporters for sodium/alanine and alanine (TransportDB [1]). According to KEGG [2], TP0414 is D-alanine and glycine permease (dagA).
L-Glutamate C00025	GTU	E0206 Predicted chemotaxis specificity includes aspartate, glutamate, histidine, glucose, and ribose [1]. E0229 Presence of transporters for glutamate (TransportDB [1]).
L-Histidine C00135	EGU	E0206 Predicted chemotaxis specificity includes aspartate, glutamate, histidine, glucose, and ribose [1]. E0210 The effects of the concentrations of dithiothreitol, molecular oxygen, and antioxidants on the in vitro replication of T. pallidum were studied [1]. The optimal dithiothreitol concentration was 0.65-1.62 mM. The optimum oxygen concentration was 3.0%. The optimum concentrations for the water-soluble antioxidants cobalt chloride, cocarboxylase (e.g. thiamin-PP), mannitol, and histidine were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication. Histidine and mannitol were the most critical. Catalase and superoxide dismutase were investigated for their ability to promote the growth in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. The medium supported the replication of T. pallidum at oxygen concentrations of 5-7%. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth in this medium.
L-Isoleucine C00407	GT	E0230 Presence of transporters for branched-chain amino acid (TransportDB [1]).
L-Leucine C00123	GT	E0230 Presence of transporters for branched-chain amino acid (TransportDB [1]).
L-Valine C00183	GT	E0230 Presence of transporters for branched-chain amino acid (TransportDB [1]).
Adenine C00147	S	E0244 No de novo purine synthetic pathway. Salvage pathway may utilize adenine, adenosine, deoxyadenosine and guanine (according to KEGG [1]).
Adenosine C00212	S	E0244 No de novo purine synthetic pathway. Salvage pathway may utilize adenine, adenosine, deoxyadenosine and guanine (according to KEGG [1]).
Deoxyadenosine C00559	S	E0244 No de novo purine synthetic pathway. Salvage pathway may utilize adenine, adenosine, deoxyadenosine and guanine (according to KEGG [1]).
Guanine C00242	S	E0244 No de novo purine synthetic pathway. Salvage pathway may utilize adenine, adenosine, deoxyadenosine and guanine (according to KEGG [1]).
Cytidine C00475	S	E0245 No de novo pyrimidine synthetic pathway. Salvage pathway may utilize uracil, cytidine, dUMP (according to KEGG [1]).
Uracil C00106	S	E0245 No de novo pyrimidine synthetic pathway. Salvage pathway may utilize uracil, cytidine, dUMP (according to KEGG [1]).
dUMP C00365	S	E0245 No de novo pyrimidine synthetic pathway. Salvage pathway may utilize uracil, cytidine, dUMP (according to KEGG [1]).
Folic acid C00504	D	E0252 There is no folate biosynthetic pathway (according to KEGG [1]). (Folate is a vitamin form of a coenzyme, tetrahydrofolate).
Biotin C00120	CD	E0251 There is no biotin biosynthetic pathway (according to KEGG [1]) but a metabolism check system (under development, H. Ogata) suggests T. pallidum requires biotin.
Nicotinamide C00153	DS	E0249 No de novo biosynthesis of nicotinate or nicotinamide, which are precursors of NAD+ and NADP+ (according to KEGG [1]).
Nicotinate C00253	DS	E0249 No de novo biosynthesis of nicotinate or nicotinamide, which are precursors of NAD+ and NADP+ (according to KEGG [1]).
CoA C00010	DS	E0250 No coenzyme A (CoA) biosynthesis (according to KEGG [1]). We may add in the medium a precursor pantetheine 4'-phosphate or CoA.
Acetyl-CoA C00024	D	E0243 A significant deficiency is the absence of fatty acid biosynthesis and degradation (according to KEGG [1]). We may add in the medium fatty acids as well as acetyl-CoA.
Pantetheine 4'-phosphate C01134	DS	E0250 No coenzyme A (CoA) biosynthesis (according to KEGG [1]). We may add in the medium a precursor pantetheine 4'-phosphate or CoA.
Pyridoxal phosphate C00018	C	E0255 Glutamate racemase (murI) [EC:5.1.1.3] is a pyridoxal-phosphate protein (according to KEGG [1]). Thus pyridoxal-phosphate may be important for Treponema. However, there is also a counter evidence. MurI activity could be independent of the cofactor [2,3].
Pyridoxine C00314	V	E0255 Glutamate racemase (murI) [EC:5.1.1.3] is a pyridoxal-phosphate protein (according to KEGG [1]). Thus pyridoxal-phosphate may be important for Treponema. However, there is also a counter evidence. MurI activity could be independent of the cofactor [2,3].
FAD C00016	CD	E0248 No synthetic pathway for riboflavin and FAD, although FAD can be generated from riboflavin (according to KEGG [1]). A metabolism check system (under development, H. Ogata) suggests T. pallidum requires FAD and FMN.
Riboflavin C00255	DV	E0248 No synthetic pathway for riboflavin and FAD, although FAD can be generated from riboflavin (according to KEGG [1]). A metabolism check system (under development, H. Ogata) suggests T. pallidum requires FAD and FMN.
Thiamin diphosphate C00068	CDER	E0205 Treponema denticola and Treponema vincentii require albumin, oleic acid, and thiamine pyrophosphate (TPP) for growth [1]. (Albumin is a generic name for soluble proteins present in the cells and the liquid moiety of animal and plant). E0210 The effects of the concentrations of dithiothreitol, molecular oxygen, and antioxidants on the in vitro replication of T. pallidum were studied [1]. The optimal dithiothreitol concentration was 0.65-1.62 mM. The optimum oxygen concentration was 3.0%. The optimum concentrations for the water-soluble antioxidants cobalt chloride, cocarboxylase (e.g. thiamin-PP), mannitol, and histidine were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication. Histidine and mannitol were the most critical. Catalase and superoxide dismutase were investigated for their ability to promote the growth in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. The medium supported the replication of T. pallidum at oxygen concentrations of 5-7%. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth in this medium. E0247 No thiamin biosynthetic pathway (according to KEGG [1]). Transketolase A (tktA) [EC:2.2.1.1] uses thiamin diphosphate as a cofactor [2].
Thiamin (vitamin B1) C00378	RT	E0205 Treponema denticola and Treponema vincentii require albumin, oleic acid, and thiamine pyrophosphate (TPP) for growth [1]. (Albumin is a generic name for soluble proteins present in the cells and the liquid moiety of animal and plant). E0217 Presence of transporters for molybdenum (TransportDB [1]). According to [2,3], the transporter is specific to thiamine.
Spermidine C00315	T	E0219 Presence of transporters for spermidine and/or putrescine (TransportDB [1]). Spermidine stabilizes DNA and RNA structure, and enhances translation processes. Spermidine is synthesized from putrescine. There is known transporter in Leishmania for putrescine and spermidine [2].
Putrescine C00134	T	E0219 Presence of transporters for spermidine and/or putrescine (TransportDB [1]). Spermidine stabilizes DNA and RNA structure, and enhances translation processes. Spermidine is synthesized from putrescine. There is known transporter in Leishmania for putrescine and spermidine [2].
vitamin E acetate	E	E0210 The effects of the concentrations of dithiothreitol, molecular oxygen, and antioxidants on the in vitro replication of T. pallidum were studied [1]. The optimal dithiothreitol concentration was 0.65-1.62 mM. The optimum oxygen concentration was 3.0%. The optimum concentrations for the water-soluble antioxidants cobalt chloride, cocarboxylase (e.g. thiamin-PP), mannitol, and histidine were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication. Histidine and mannitol were the most critical. Catalase and superoxide dismutase were investigated for their ability to promote the growth in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. The medium supported the replication of T. pallidum at oxygen concentrations of 5-7%. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth in this medium.
D-Ribose C00121	TU	E0206 Predicted chemotaxis specificity includes aspartate, glutamate, histidine, glucose, and ribose [1]. E0216 Presence of transporters for sugar (TransportDB [1]), which include ribose, galactose, malate, succinate, glucose, glycerol-P [2].
Choline C00114	ST	E0202 Glucose is incorporated only in pentose of nucleic acids, aspartate in proteins, glycerol of phosphatidylcholin and phosphatidylglycerol [1,2]. Exogenous choline was incorporated in phosphatidylcholin [2]. E0226 Presence of transporters for carnitine/choline (TransportDB [1]). (Carnitine is highly concentrated in animal testes.)
D-Alanine C00133	T	E0225 Presence of transporters for sodium/alanine and alanine (TransportDB [1]). According to KEGG [2], TP0414 is D-alanine and glycine permease (dagA).
Iron-sulfur C00824	C	E0253 Ribonucleoside-diphosphate reductase [EC:1.17.4.1] uses iron and ATP as cofactors [1]. Glutamate synthase (NADPH) [EC:1.4.1.13] uses iron-sulfur as a cofactor [2].
Oleate C00712	R	E0205 Treponema denticola and Treponema vincentii require albumin, oleic acid, and thiamine pyrophosphate (TPP) for growth [1]. (Albumin is a generic name for soluble proteins present in the cells and the liquid moiety of animal and plant).
Carnitine C00487	TU	E0226 Presence of transporters for carnitine/choline (TransportDB [1]). (Carnitine is highly concentrated in animal testes.)
Iron (Fe2+,Fe3+) C00023	CN	E0253 Ribonucleoside-diphosphate reductase [EC:1.17.4.1] uses iron and ATP as cofactors [1]. Glutamate synthase (NADPH) [EC:1.4.1.13] uses iron-sulfur as a cofactor [2].
Copper (Cu2+) C00070	NT	E0224 Presence of transporters for copper (TransportDB [1]).
Manganese (Mn2+) C00034	CNT	E0215 Presence of transporters for manganese (TransportDB [1]). Oxaloacetate decarboxylase [EC:4.1.1.3] utilizes manganese as well as sodium as cofactors [2].
Molybdenum (Mo) C00150	NT	E0217 Presence of transporters for molybdenum (TransportDB [1]). According to [2,3], the transporter is specific to thiamine.
Cobalt (Co2+) C00175	CNT	E0222 Presence of transporters for cobalt (TransportDB [1]). There is a enzyme [EC:1.1.1.267] requiring cobalt as a cofactor.
Chromium (Cr) C06268	NT	E0228 Presence of transporters for chromium (TransportDB [1]).
cobalt chloride	E	E0210 The effects of the concentrations of dithiothreitol, molecular oxygen, and antioxidants on the in vitro replication of T. pallidum were studied [1]. The optimal dithiothreitol concentration was 0.65-1.62 mM. The optimum oxygen concentration was 3.0%. The optimum concentrations for the water-soluble antioxidants cobalt chloride, cocarboxylase (e.g. thiamin-PP), mannitol, and histidine were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication. Histidine and mannitol were the most critical. Catalase and superoxide dismutase were investigated for their ability to promote the growth in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. The medium supported the replication of T. pallidum at oxygen concentrations of 5-7%. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth in this medium.
Calcium (Ca2+) C00076	NT	E0227 Presence of transporters for sodium/calcium (TransportDB [1]).
H+ C00080	NT	E0223 Presence of transporters for H+ or Na+ (TransportDB [1]).
Potassium (K+) C00238	NT	E0231 Presence of transporters for sodium/potassium and potassium (TransportDB [1]).
Magnesium (Mg2+) C00305	CN	E0254 Enolase [EC:4.2.1.11] uses magnesium as a cofactor [1].
Sodium (Na+) C01330	CNT	E0215 Presence of transporters for manganese (TransportDB [1]). Oxaloacetate decarboxylase [EC:4.1.1.3] utilizes manganese as well as sodium as cofactors [2]. E0223 Presence of transporters for H+ or Na+ (TransportDB [1]). E0227 Presence of transporters for sodium/calcium (TransportDB [1]). E0231 Presence of transporters for sodium/potassium and potassium (TransportDB [1]).
ox serum	E	E0208 T. pallidum was incubated with and without cells in cell culture medium (pH 7.3), under deoxygenated conditions. Five to ten percent of the treponemes remained motile in cell-treponeme systems of co-incubation. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Continuous passage of the T. pallidum was not achieved in vitro [1]. A nonpathogenic treponeme were maintained and passaged in thioglycolate medium with 10% heat-inactivated normal rabbit serum at 37 degree C [2].
rabbit serum	E	E0208 T. pallidum was incubated with and without cells in cell culture medium (pH 7.3), under deoxygenated conditions. Five to ten percent of the treponemes remained motile in cell-treponeme systems of co-incubation. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Continuous passage of the T. pallidum was not achieved in vitro [1]. A nonpathogenic treponeme were maintained and passaged in thioglycolate medium with 10% heat-inactivated normal rabbit serum at 37 degree C [2].
albumin	R	E0205 Treponema denticola and Treponema vincentii require albumin, oleic acid, and thiamine pyrophosphate (TPP) for growth [1]. (Albumin is a generic name for soluble proteins present in the cells and the liquid moiety of animal and plant).
Fatty acid C00162	DGT	E0203 Treponema pallidum (and Borrelia burgdorferi) incorporates directory fatty acids into the outer membrane [1], because of the lack of lipopolysaccharide in the outer membrane. Gram-negative bacteria exhibit poor permeability of hydrophobic compounds like fatty acids. E0243 A significant deficiency is the absence of fatty acid biosynthesis and degradation (according to KEGG [1]). We may add in the medium fatty acids as well as acetyl-CoA.
Dithiothreitol C00265	E	E0201 T. pallidum is microaerophilic, which is sensitive but require oxygen (O2). The addition of sodium metabisulfite to a Eagle's minimal essential medium with rabbit serum was found to have an effect similar to that of dithiothreitol in the survival of T. pallidum under 3% O2. Analyses revealed that 50% motility was retained longest at O2 concentrations of 1-5%, whether or not dithiothreitol or sodium metabisulfite were present. Concentrations of O2 of 3-10% were optimal for nucleic acid synthesis. Sodium metabisulfite was less effective than dithiothreitol in stimulating nucleic acid synthesis. During incubation under 3% O2, motility was maintained at > 50% for 15 days. The optimal retention of motility and nucleic acid synthesis under microaerobic conditions in the absence of reducing compounds provides evidence that T. pallidum is a microaerophilic organism [1]. E0210 The effects of the concentrations of dithiothreitol, molecular oxygen, and antioxidants on the in vitro replication of T. pallidum were studied [1]. The optimal dithiothreitol concentration was 0.65-1.62 mM. The optimum oxygen concentration was 3.0%. The optimum concentrations for the water-soluble antioxidants cobalt chloride, cocarboxylase (e.g. thiamin-PP), mannitol, and histidine were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication. Histidine and mannitol were the most critical. Catalase and superoxide dismutase were investigated for their ability to promote the growth in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. The medium supported the replication of T. pallidum at oxygen concentrations of 5-7%. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth in this medium.
sodium metabisulfite	E	E0201 T. pallidum is microaerophilic, which is sensitive but require oxygen (O2). The addition of sodium metabisulfite to a Eagle's minimal essential medium with rabbit serum was found to have an effect similar to that of dithiothreitol in the survival of T. pallidum under 3% O2. Analyses revealed that 50% motility was retained longest at O2 concentrations of 1-5%, whether or not dithiothreitol or sodium metabisulfite were present. Concentrations of O2 of 3-10% were optimal for nucleic acid synthesis. Sodium metabisulfite was less effective than dithiothreitol in stimulating nucleic acid synthesis. During incubation under 3% O2, motility was maintained at > 50% for 15 days. The optimal retention of motility and nucleic acid synthesis under microaerobic conditions in the absence of reducing compounds provides evidence that T. pallidum is a microaerophilic organism [1].
Thioglycolate C02086	E	E0208 T. pallidum was incubated with and without cells in cell culture medium (pH 7.3), under deoxygenated conditions. Five to ten percent of the treponemes remained motile in cell-treponeme systems of co-incubation. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Continuous passage of the T. pallidum was not achieved in vitro [1]. A nonpathogenic treponeme were maintained and passaged in thioglycolate medium with 10% heat-inactivated normal rabbit serum at 37 degree C [2].
sodium thioglycolate	E	E0208 T. pallidum was incubated with and without cells in cell culture medium (pH 7.3), under deoxygenated conditions. Five to ten percent of the treponemes remained motile in cell-treponeme systems of co-incubation. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Continuous passage of the T. pallidum was not achieved in vitro [1]. A nonpathogenic treponeme were maintained and passaged in thioglycolate medium with 10% heat-inactivated normal rabbit serum at 37 degree C [2].
catalase EC:1.11.1.6	E	E0210 The effects of the concentrations of dithiothreitol, molecular oxygen, and antioxidants on the in vitro replication of T. pallidum were studied [1]. The optimal dithiothreitol concentration was 0.65-1.62 mM. The optimum oxygen concentration was 3.0%. The optimum concentrations for the water-soluble antioxidants cobalt chloride, cocarboxylase (e.g. thiamin-PP), mannitol, and histidine were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication. Histidine and mannitol were the most critical. Catalase and superoxide dismutase were investigated for their ability to promote the growth in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. The medium supported the replication of T. pallidum at oxygen concentrations of 5-7%. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth in this medium.
superoxide dismutase EC:1.15.1.1	E	E0210 The effects of the concentrations of dithiothreitol, molecular oxygen, and antioxidants on the in vitro replication of T. pallidum were studied [1]. The optimal dithiothreitol concentration was 0.65-1.62 mM. The optimum oxygen concentration was 3.0%. The optimum concentrations for the water-soluble antioxidants cobalt chloride, cocarboxylase (e.g. thiamin-PP), mannitol, and histidine were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication. Histidine and mannitol were the most critical. Catalase and superoxide dismutase were investigated for their ability to promote the growth in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. The medium supported the replication of T. pallidum at oxygen concentrations of 5-7%. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth in this medium.
Retinol (vitamin A) C00473	E	E0210 The effects of the concentrations of dithiothreitol, molecular oxygen, and antioxidants on the in vitro replication of T. pallidum were studied [1]. The optimal dithiothreitol concentration was 0.65-1.62 mM. The optimum oxygen concentration was 3.0%. The optimum concentrations for the water-soluble antioxidants cobalt chloride, cocarboxylase (e.g. thiamin-PP), mannitol, and histidine were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication. Histidine and mannitol were the most critical. Catalase and superoxide dismutase were investigated for their ability to promote the growth in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. The medium supported the replication of T. pallidum at oxygen concentrations of 5-7%. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth in this medium.
Fibronectin C00516	U	E0207 Pathogenic Treponema bind host fibronectin to penetrate organs [1].