Biosynthetic ability of diverse basidiomycetous yeast strains to produce the natural antioxidant ergothioneine.

Sixteen strains of basidiomycetous yeasts were evaluated for their capability to produce ergothioneine (EGT), an amino acid derivative with strong antioxidant activity. The cells were cultured in either two synthetic media or yeast mold (YM) medium for 72 h, after which cytosolic constituents were extracted from the cells with hot water. After analyzing the extracts via liquid chromatography-mass spectrometry (LC-MS), we found that all strains produced varying amounts of EGT. The EGT-producing strains, including Ustilago siamensis, Anthracocystis floculossa, Tridiomyces crassus, Ustilago shanxiensis, and Moesziomyces antarcticus, were subjected to flask cultivation in YM medium. U. siamensis CBS9960 produced the highest amount of EGT at 49.5 ± 7.0 mg/L after 120 h, followed by T. crassus at 30.9 ± 1.8 mg/L. U. siamensis was also cultured in a jar fermenter and produced slightly higher amounts of EGT than under flask cultivation. The effects of culture conditions, particularly the addition of precursor amino acids, on EGT production by the selected strains were also evaluated. U. siamensis showed a 1.5-fold increase in EGT production with the addition of histidine, while U. shanxiensis experienced a 1.8-fold increase in EGT production with the addition of methionine. These results suggest that basidiomycetous yeasts could serve an abundant source for natural EGT producers.

A New Screening Approach for Glycolipid-type Biosurfactant Producers Using MALDI-TOF/MS.

We applied matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) to screen for glycolipid-type biosurfactant (BS) producers. A crude extract of Pseudozyma antarctica, a well-known mannosylerythritol lipid (MEL) producer, was initially subjected to MALDI-TOF/MS. The spectrum of the extract showed the accumulation of diacylated MELs in culture. We then screened 80 environmental samples for BS-producing yeasts, and extracts from broth cultures of the selected five strains were examined using MALDI-TOF/MS. The results showed that all five strains produced MELs, whereas four strains also produced cellobiose lipids (CLs). By D1/D2 region sequence analysis, the MEL-producing strain was assigned to P. antarctica while the four MEL- and CL-producing strains were assigned to P. hubeiensis. These results demonstrate that MALDI-TOF/MS is a rapid and reliable tool to detect BS molecules in crude extracts of broth cultures to screen for glycolipid-type BS producers.