Abstract
Raman spectroscopy is a universal method designed for the analysis of a wide range of physical, chemical and biological systems or various surfaces. This technique is suitable to monitor various components of cells, tissues or microorganisms. The advantages include very fast non-contact and non-destructive analysis and no or minimal need for sample treatment. The yeasts Metschnikowia can be considered as industrially usable producers of pulcherrimin or single-cell lipids, depending on cultivation conditions and external stress. In the present study, Raman spectroscopy was used as an effective tool to identify both pulcherrimin and lipids in single yeast cells. The analysis of pulcherrimin is very demanding; so far, there is no optimal procedure to analyze or identify this pigment. Based on results, the strong dependence of pulcherrimin production on the ferric ion concentration was found with the highest yield in media containing 0.1 g/L iron. Further, production of lipids in Metschnikowia cells was studied at different temperatures and C:N ratios, using Raman spectroscopy to follow fatty acids composition, under different regimes, by monitoring the iodine number. The results of Raman spectroscopy were comparable with the fatty acid analysis obtained by gas chromatography. This study therefore supported use of Raman spectroscopy for biotechnological applications as a simple tool in the identification and analysis both the pulcherrimin and microbial lipids. This method provides a quick and relatively accurate estimation of targeted metabolites with minimal sample modification and allows to monitor metabolic changes over time of cultivation.
Highlights
Pulcherrimin potency to exert antagonistic effects of Metschnikowia yeasts correlates with the amount of pulcherrimin production
The antimicrobial activity occurs when iron immobilizing pulcherrimin, together with pulcherriminic acid, sequesters any iron in the environment which, in turn, prevents other microorganisms’ growth, because iron is an element vital for the growth
We aim to explore pulcherrimin production characteristics on selected samples when yeast is cultivated in the presence of different concentrations of ferric ions
Summary
Introduction published maps and institutional affilRaman spectroscopy is a spectroscopic technique with many types of applications; it is commonly used to provide a structural fingerprint and composition of substances [1,2].The method is based on inelastic scattering of photons, known as Raman scattering, that allows identification of exact molecular spectrum of biological/non-biological sample [3,4,5,6].Conventional analytical techniques, such as gas chromatography or high-performance liquid chromatography, are commonly used to detect different metabolites or lipids [7,8].these conventional technologies are destructive, time-consuming and often expensive. The method is based on inelastic scattering of photons, known as Raman scattering, that allows identification of exact molecular spectrum of biological/non-biological sample [3,4,5,6]. Conventional analytical techniques, such as gas chromatography or high-performance liquid chromatography, are commonly used to detect different metabolites or lipids [7,8]. These conventional technologies are destructive, time-consuming and often expensive. The main goal of our investigations is the development of specialized instrumentation and methodology to monitor metabolic states of microorganisms iations
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