RESUMO
The extracellular slime substance produced by Staphylococcus epidermidis was investigated. Slime production was assessed by bacterial agglutination in the presence of concanavalin A (Con A) or poly-L-lysine and by bacterial adherence to polyethylene. Media for slime production was optimized using these criteria. A phenol-saline extract of crude slime was separated into four fractions on a DEAE-sepharose column. Total protein and sugar content and the monosaccharide constituents were determined. Crude slime and the phenol-saline extract showed a strong precipitation reaction with Con A and poly-L-lysine (double diffusion). Fractions I and II containing mannose as the most abundant sugar reacted with Con A and two other mannose-specific lectins (Lens culinaris, Pisum sativum). This reaction could be inhibited by mannose. Fractions III and IV were precipitated by poly-L-lysine, probably due to a reaction with glucuronic acid which was only present in these fractions. Precoating of polyethylene with crude slime, phenol-saline extract and fractions III and IV resulted in a marked inhibition of attachment of staphylococcal cells. Production of the extracellular slime substance was completely inhibited by subinhibitory concentrations of the glycosylation inhibitor tunicamycin, whereas penicillin had no influence. Extracellular slime substance produced by S. epidermidis seems to be a complex of glycoconjugate character and plays an important role in the attachment to synthetic polymers. The production of slime by staphylococci can be easily determined using mannose specific lectins and poly-L-lysine.
Assuntos
Glicoproteínas/análise , Lipoproteínas/análise , Staphylococcus epidermidis/análise , Adesividade , Testes de Aglutinação , Proteínas de Bactérias/análise , Concanavalina A/farmacologia , Meios de Cultura , Glicoproteínas/biossíntese , Glicoproteínas/imunologia , Imunodifusão , Lipoproteínas/biossíntese , Lipoproteínas/imunologia , Penicilinas/farmacologia , Polietilenos , Polilisina/farmacologia , Staphylococcus epidermidis/imunologia , Staphylococcus epidermidis/metabolismo , Tunicamicina/farmacologiaRESUMO
Tissue slices from the liver and brain of 7-day-old rats incubated with [1-14C]stearic acid desaturate the stearate to oleate. The activities of the two tissues are different but of the same order of magnitude. With increasing age, the activity in the liver increases markedly, while the brain activity decreases. The postmitochondrial supernatant from adult (3-month-old) liver contains 2 to 3 orders of magnitude more stearoyl-CoA dehydrogenase activity than the brain postmitochondrial fraction. The washed microsomal fraction from liver had about the same activity as the postmitochondrial supernatant, but no dehydrogenase activity could be detected in the washed microsomal fraction from the brain. The acyl-CoA synthetase and the palmitoyl-CoA hydrolase activities measured in the washed microsomes from adult brain were both lower than in liver microsomes. The concentration of stearoyl-CoA (the substrate for the stearoyl-CoA dehydrogenase) resulting from the ratio of these activities was too high, however, for the lack of desaturase activity to have been simulated by lack of substrate.