title: Albumin requirement of Treponema denticola and Treponema vincentii.
abstract: Treponema denticola and Treponema vincentii were found to require albumin, oleic acid, and thiamine pyrophosphate (TPP) for growth. Previous studies indicated that commercial human alpha globulin, which is 50% albumin, was the only serum fraction that supported growth of these two oral treponemes. The alpha-globulin proteins were separated from the contaminating albumin with Affi-Gel Blue affinity chromatography. Both the albumin fraction and one of the alpha-globulin fractions were required for growth of T. denticola. Oleic acid was supplied by the alpha-globulin fraction and the albumin functioned as a chelator to maintain a low level of free fatty acid in the medium. Purified serum albumin (bovine or human) could substitute for the alpha-globulin fraction that contained albumin. Optimal growth of T. denticola and T. vincentii was in a medium supplemented with 0.4% (w/v) delipified albumin, 0.08 mg/mL of sodium oleate, and 25 micrograms/mL of TPP. Serum albumin tightly bound TPP (0.5 microgram of TPP per milligram of albumin). Optimal growth of T. denticola was only in an albumin-oleate supplemented medium with sufficient TPP to saturate the albumin binding sites and provide excess free TPP. Albumin bound long-chain fatty acids and thus detoxified the medium. Neither starch- nor charcoal-treated Tween 80 (polysorbitan monooleate) replaced albumin for optimal growth. Short-chain fatty acids supported only limited growth of T. denticola when added to a medium with TPP or to a medium that contained 0.4% delipified albumin and TPP.
mesh_terms: D000818:Animals; D002417:Cattle; D003470:Culture Media; D006801:Humans; D007700:Kinetics; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013045:Species Specificity; D013835:Thiamine Pyrophosphate / Q000494:pharmacology; D014208:Treponema / Q000187:drug effects / Q000254:growth & development*
chemical_list: D003470:Culture Media; D009829:Oleic Acids; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D019301:Oleic Acid; D013835:Thiamine Pyrophosphate