Lipolytic enzymes of Trichophyton rubrum
TL;DR: Supernatant obtained after removal of 1,005,000 g sedimentable fragments from cell extract contains acyl CoA lysolecithin acyl transferase which requires ATP, CoA, Mg2+ and an unsaturated fatty acid for its activity.
Abstract: Trichophyton rubrum cells contain lipase, phospholipases A and B and acyl CoA lysolecithin acyl transferase activities. This dermatophyte excretes lipase and phospholipase A into the growth medium when cultivated in Sabouraud's broth. Extracellular lipase has optimum activity at pH 8.0 whereas the intracellular lipase is maximally active at pH 8.0 whereas the intracellular lipase is maximally active at pH 7.0. The optimum pH of phospholipase A and B activities which are localized in 15000 g sedimentable cell fragments are 7.0 and 6.0 respectively. Supernatant obtained after removal of 1,005,000 g sedimentable fragments from cell extract contains acyl CoA lysolecithin acyl transferase which requires ATP, CoA, Mg2+ and an unsaturated fatty acid for its activity.
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TL;DR: An exocellular proteinase produced by Trichophyton rubrum in a glucose-peptone broth was purified from lyophilized and dialysed culture filtrate of the dermatophyte and showed an alkaline pH optimum and was not activated by divalent metal ions but inhibited strongly by phenylmethylsulfonylfluoride.
Abstract: An exocellular proteinase produced by Trichophyton rubrum in a glucose-peptone broth was purified from lyophilized and dialysed culture filtrate of the dermatophyte by Sephadex G-100 gel filtration and preparative polyacrylamide gel electrophoresis. The purified enzyme was a homogeneous protein of molecular weight 34 700 and it could hydrolyse azoalbumin, casein, bovine serum albumin, α-N-benzoyl-L-arginine ethyl ester and p-toluenesulfonyl-L-arginine methyl ester but not N-benzoyl-L-tyrosine ethyl ester, α-N-benzoyl-DL-arginine-p-nitroanilide and keratin. The enzyme showed an alkaline pH optimum and was not activated by divalent metal ions but inhibited strongly by phenylmethylsulfonylfluoride. Thus the enzyme was identified as an alkaline serine proteinase.
70 citations
TL;DR: In this paper, the authors determined skin barrier function, epidermal differentiation, and human-β-defensin 2 (hBD-2) protein expression in 10 patients with tinea corporis caused by Trichophyton rubrum (T. rubrum).
70 citations
64 citations
TL;DR: Alkaline phosphatase, esterases and leucine arylamidase may be important for the parasitic growth of dermatophytes and may be helpful for species identification.
Abstract: Summary. Exoenzymes produced by common dermatophytes, in addition to their ability to cause cutaneous inflammation, are thought to contribute to fungal spread. To investigate the patterns of enzymes released by common dermatophytes as well as Scopulariopsis brevicaulis, the fungi were grown in liquid media containing either hair, stratum corneum, neopeptone or lipids, or in RPMI medium. Enzymes recovered from the culture supernatants were compared using the Apizym test. As a result, the widest range of enzymes was seen in protein-containing media, with a maximum of 13 enzymes stimulated by growth on hair. Dermatophytes in all protein media produced high levels of alkaline phosphatase, esterases and leucine arylamidase. In these media the highest total enzymatic activity was released by Microsporum canis, whereas the lowest was released by Epidermophyton floccosum. Although RPMI broth stimulated luxuriant growth of all species, recovery was limited to a maximum of six enzymes. In lipid medium E. floccosum and M. canis failed to grow. When comparing the various nutrients, Scopulariopsis released fewer enzymes than the dermatophytes and only minor quantities of alkaline phosphatase. We conclude that alkaline phosphatase, esterases and leucine arylamidase may be important for the parasitic growth of dermatophytes. The total enzymatic activity of dermatophytes appears to be correlated with the intensity of cutaneous inflammation. Furthermore, enzyme measurements may be helpful for species identification.
Zusammenfassung. Exoenzyme von Dermatophyten tragen sowohl zum Wachstum der Pilze in der Haut bei als auch zur kutanen Entzundungsreaktion. Um die Enzymmuster haufiger Dermatophyten und von Scopulariopsis brevicaulis zu bestimmen, wurden die Erreger in Nahrlosungen mit entweder Haar, Stratum corneum, Neopepton oder Fetten, sowie in RPMI-Medium gezuchtet. Die Enzyme in den Kulturuberstanden wurden mit Hilfe des Api-zymR Tests verglichen. Die breitesten Spektren mit bis zu 13 Enzymen wurden von Haaren induziert, gefolgt von den anderen Proteinsubstraten. In diesen Medien wurden fur alle Dermatophyten hohe Aktivitaten von alkalischer Phosphatase, Esterasen und Leucinarylamidase gemessen. Die hochste Gesamtenzymaktivitat in den Proteinmedien wurde von Microsporum canis, die geringste von Epidermophyton floccosum freigesetzt. Alle Arten wuchsen gut in RPMI-Medium, wenngleich dabei maximal nur 6 Enzyme nachweisbar waren. Im Lipidmedium wuchsen E. floccosum und M. canis nicht. Unter Berucksichtigung aller Nahrmedien setzte Scopulariopsis insgesamt eine geringere Zahl von Enzymen frei als die Dermatophyten und nur eine niedrige Aktivitat von alkalischer Phosphatase. Nach unseren Ergebnissen konnten besonders alkalische Phosphatase, Esterasen und Leucinarylamidase fur das parasitare Wachstum von Dermatophyten bedeutsam sein. Die Enzymaktivitat von Dermatophyten scheint mit der Intensitat der kutanen Entzundung zu korrelieren. Enzymmusterbestimmungen konnten auserdem hilfreich bei der Artdifferenzierung sein.
45 citations
References
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Journal Article•
TL;DR: Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein.
289,852 citations
TL;DR: In this article, thin-layer chromatography was used to quantify all lipid classes to a lower limit of 15 μg of lipid, where the reduction in absorbance at 350 mμ is proportional to the amount of lipid.
369 citations
TL;DR: Trichophyton rubrum grown in Sabouraud's liquid medium contains phosphatidyl inositol, polyphosphatids inositols, and the relative proportion of these components in total phospholipid fraction remains almost constant throughout the culture life of this fungus.
Abstract: Trichophyton rubrum grown in Sabouraud's liquid medium contains phosphatidyl inositol, polyphosphatidyl inositol, phosphatidyl choline, phosphatidyl serine, phosphatidyl ethanolamine phosphatidyl glycerol and phosphatidic acid in the polar lipid fraction of its mycelia. The relative proportion of these components in total phospholipid fraction remains almost constant throughout the culture life of this fungus.
23 citations
TL;DR: Among subcultured strains, it was found a decreased lipolytic activity in Microsporum species and an increased lipolyic activity in Epidermophyton and someTrichophyton species.
Abstract: Lipolytic activity of dermatophytes was tested by the method generally used forCandida lipolytica. Most of the freshly isolated strains ofMicrosporum canis, M. gypseum, Epidermophyton floccosum andTrichophyton mentagrophytes gave positive reactions, whereas, only few strains ofT. schoenleini, T. violaceum, T. megnini, T. rubrum andT. tonsurans yielded such reactions. Among subcultured strains, it was found a decreased lipolytic activity inMicrosporum species and an increased lipolytic activity inEpidermophyton and someTrichophyton species.
19 citations
TL;DR: Phospholipid synthesis, interconversion and breakdown in T. rubrum were followed by radioactive tracer, and their conversions to phosphatidyl choline and phosph atidyl inositol are suspected.
Abstract: Phospholipid synthesis, interconversion and breakdown in T. rubrum were followed by radioactive tracer. Synthesis and catabolism of phosphatidyl ethanolamine and phosphatidyl serine are most rapid; phosphatidyl choline and phosphatidyl inositol are metabolised rather slowly. Catabolism of phosphatidyl ethanolamine and phosphatidyl serine are uniform and their conversions to phosphatidyl choline and phosphatidyl inositol are suspected.
10 citations