Use of high-throughput techniques to study simultaneous production of lipid metabolites in carotenogenic yeasts grown on waste animal fat
I. Marova, M. Rapta, M. Vanek, A. Haronikova, M. Szotkowski, V. Shapaval
Characterization of metabolic activity of yeasts in biotechnological applications is an important issue both for routine analysis and trouble-shooting incidences. In this study we present results of high-throughput micro-cultivation, FTIR spectroscopy and single cell analysis of industrial red yeasts. As a substrate waste animal fat and its hydrolysis products were used according to the biorefinery concept.
Twenty strains of red yeasts (Rhodotorula, Sporobolomyces, Cystofilobasidium) were used for analyses. As the substrate hydrolyzed and crude animal was used. Yeasts were cultivated in medium with particular C/N ratio. Simultaneously with growth characteristics total metabolic activity (FTIR) and production of lipids and fatty acids (GC/FID), sterols, carotenoids and CoQ (LC/MS) were followed. Glycerol, lipase and lipids were measured by enzyme methods. Cells were monitored by advanced fluorescence techniques on single-cell level.
Optimal medium composition and C/N ratio was found by FTIR. All strains were able to utilize fat hydrolyzate, some of them produce lipase and utilize crude fat. Amount of lipid metabolites depended on glycerol content in medium and C/N ratio. Using time-differentiated fluorescence localization and trafficking of pigments and lipids in yeast cells was determined. The results could contribute to better understanding and control of biotechnological process.