Isolation and characterization of thermotolerant ethanol-fermenting yeasts from Laos and application of whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) analysis for their quick identification

Thermotolerant yeasts, which are expected to be applicable for high-temperature fermentation as an economical process, were isolated from four provinces in Laos. Of these yeasts, five isolates exhibited stronger fermentation abilities in a 16% sugars-containing medium of glucose, sucrose, sugarcane or molasses at 40°C than that of Kluyveromyces marxianus DMKU 3-1042, one of the most thermotolerant and efficient yeasts isolated previously in Thailand. One of the five strains, BUNL-17, exhibited the highest ethanol fermentation performance at 45°C. Yeast identification was achieved by whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) analysis as well as by nucleotide sequencing of the D1/D2 domain of the large subunit rRNA gene, revealing that the isolated strains can be categorized into Pichia kudriavzevii, Cyberlindnera rhodanensis and K. marxianus and that all of the five strains are K. marxianus. The results of this study showed that the former analysis is much faster than the latter and reliable and equivalent to the latter. Key words: Ethanol fermentation, thermotolerant yeast, Kluyveromyces marxianus.