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Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins
Published on: June 20, 2019
Hydrolase activity of 150 Malassezia furfur isolates of different clinical origin
P Mayser1, C Scheurer, C Papavassilis
1Department of Dermatology and Andrology, Justus-Liebig-University, Giessen, FR Germany.
Abstract:
Apart from pityriasis versicolor, Malassezia furfur is thought to play a significant role in the pathogenesis of seborrhoic eczema and Malassezia folliculitis. However, it has not been clarified whether in addition to host factors (e.g. immune status, greasy skin), yeast-dependent activities are responsible for manifestation of the disease. In this context interstrain variability of hydrolase activity of Malassezia isolates might be significant. For determination of hydrolase activity, washed yeast suspension was applied to selective agar for pathogenic fungi containing 8% (v/v) Tween 80 or Tween 60 and was incubated at 37 degrees C for 10 days. Growth was accompanied by formation of a dense white zone around the colony, in which free fatty acids corresponding to Tween 80 (C18:1) or Tween 60 (16:0, 18:0) were demonstrated by means of thin layer and gas chromatography. Thus, this phenomenon is a parameter for yeast-dependent hydrolysis of Tween 80 and Tween 60. Considering different growth behaviour, a "hydrolase zone' (H2) was determined using the quotient colony diameter/(ring+colony) diameter in each of the 150 strains tested. Although no significant H2 variations were observed in strains of different clinical origin, the present study revealed that in addition to a known enzyme, which is located within the cell wall and/or membrane systems, these Malassezia isolates produce a very active hydrolase diffusing into the medium.
Insights
Malassezia furfur yeast isolates exhibit significant hydrolase activity, contributing to skin conditions like seborrheic eczema and folliculitis. This study quantifies yeast-dependent hydrolase activity, revealing a diffusible enzyme in Malassezia strains.
Area of Science:
- Medical Mycology
- Dermatology
- Biochemistry
Background:
- Malassezia furfur is implicated in pityriasis versicolor, seborrheic eczema, and Malassezia folliculitis.
- The role of yeast-dependent activities versus host factors in disease pathogenesis remains unclear.
- Interstrain variability in hydrolase activity may be crucial for disease manifestation.
Purpose of the Study:
- To investigate the hydrolase activity of Malassezia isolates.
- To determine if yeast-dependent activities contribute to skin disease pathogenesis.
- To assess interstrain variability in hydrolase activity.
Main Methods:
- 150 Malassezia strains were tested for hydrolase activity on selective agar with Tween 80 or Tween 60.
- Incubation at 37°C for 10 days.
- Hydrolase activity was quantified by measuring the "hydrolase zone" (H2) and confirmed using thin layer and gas chromatography.
Main Results:
- A distinct white zone around colonies indicated yeast-dependent hydrolysis of Tween 80 and Tween 60.
- Free fatty acids were identified, confirming the hydrolysis.
- No significant H2 variations were found across strains from different clinical origins.
- Malassezia isolates produce a highly active hydrolase that diffuses into the medium, beyond cell-associated enzymes.
Conclusions:
- Malassezia isolates possess a diffusible, highly active hydrolase.
- This enzyme activity likely contributes to the pathogenesis of Malassezia-related skin diseases.
- Further research is needed to fully elucidate the role of this hydrolase in specific conditions.
