Growth inhibition of Candida species by Wickerhamomyces anomalus mycocin and a lactone compound of Aureobasidium pullulans.
BACKGROUND
The increasing resistance of Candida yeasts towards antifungal compounds and the limited choice of therapeutic drugs have spurred great interest amongst the scientific community to search for alternative anti-Candida compounds. Mycocins and fungal metabolites have been reported to have the potential for treatment of fungal infections. In this study, the growth inhibition of Candida species by a mycocin produced by Wickerhamomyces anomalus and a lactone compound from Aureobasidium pullulans were investigated.


METHODS
Mycocin was purified from the culture supernatant of an environmental isolate of W. anomalus using Sephadex G-75 gel filtration column chromatography. The mycocin preparation was subjected to SDS-PAGE analysis followed by MALDI TOF/TOF mass spectrometry analysis. The thermal and temperature stability of the mycocin were determined. The glucanase activity of the mycocin was investigated by substrate staining of the mycocin with 4-methyl-umbelliferyl-ß-D-glucoside (MUG). Gas chromatography mass spectrometry (GCMS) analysis was used to identify anti-Candida metabolite in the culture supernatant of an environmental isolate of Aureobasidium pullulans. The inhibitory effects of the anti-Candida compound against planktonic and biofilm cultures of various Candida species were determined using broth microdilution and biofilm quantitation methods.


RESULTS
A mycocin active against Candida mesorugosa but not C. albicans, C. parapsilosis and C. krusei was isolated from the culture supernatant of W. anomalus in this study. The mycocin, identified as exo-ß-1,3 glucanase by MALDI TOF/TOF mass spectrometry, was stable at pH 3-6 and temperature ranging from 4-37°C. The glucanase activity of the mycocin was confirmed by substrate staining with MUG. 5-hydroxy-2-decenoic acid lactone (HDCL) was identified from the culture supernatant of A. pullulans. Using a commercial source of HDCL, the planktonic and biofilm MICs of HDCL against various Candida species were determined in this study.


CONCLUSIONS
W. anomalus mycocin demonstrated a narrow spectrum of activity targeting only against C. mesorugosa, while HDCL demonstrated a broad spectrum of inhibitory action against multiple Candida species. The growth inhibition of W. anomalus mycocin and the lactone compound from A. pullulans against Candida yeasts should be further explored for therapeutic potentials against candidiasis.
Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. tayst@um.edu.my.
Sun-Tee
ST
Tay
Su-Lin
SL
Lim
Hui-Wee
HW
Tan
D000935:Antifungal Agents / Q000302:isolation & purification / Q000494:pharmacology*; D001203:Ascomycota / Q000737:chemistry*; D018441:Biofilms / Q000187:drug effects; D001688:Biological Products / Q000302:isolation & purification / Q000494:pharmacology*; D002175:Candida / Q000187:drug effects* / Q000254:growth & development; D005229:Fatty Acids, Monounsaturated / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D007783:Lactones / Q000302:isolation & purification / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D009183:Mycotoxins / Q000302:isolation & purification / Q000494:pharmacology*; D013045:Species Specificity
D000935:Antifungal Agents; D001688:Biological Products; D005229:Fatty Acids, Monounsaturated; D007783:Lactones; D009183:Mycotoxins; C052476:2-decenoic acid