Adam Dobrowolski

Efficient conversion of crude glycerol from various industrial wastes into single cell oil by yeast Yarrowia lipolytica

In this study, crude glycerol from various industries was used to produce lipids via wild type Yarrowia lipolytica A101. We tested samples without any prior purification from five different waste products; each contained various concentrations of glycerol (42-87%) as the sole carbon source. The best results for lipid production were obtained for medium containing glycerol from fat saponification. This reached 1.69gL(-1) (25% of total cell dry weight) with a biomass yield of 0.17gg(-1) in the flasks experiment. The batch cultivation in a bioreactor resulted in enhanced lipid production-it achieved 4.72gL(-1) with a biomass yield 0.21gg(-1). Moreover, the properly selected batch of crude glycerol provides a defined fatty acid composition. In summary, this paper shows that crude glycerol from soap production could be efficiently converted to single cell oil without any prior purification.

A two-stage fermentation process of erythritol production by yeast Y. lipolytica from molasses and glycerol.

In this study, a two-stage fermentation process of erythritol production based on molasses and glycerol was investigated. During the first stage, the biomass of Yarrowia lipolytica was grown on medium containing sucrose as the sole carbon source. In the second stage, production of erythritol was initiated by glycerol addition. To use molasses as a substrate for erythritol synthesis, sucrose utilization was established by expressing the Saccharomyces cerevisiae SUC2 gene. In this study, cultivation of yeast Y. lipolytica could produce 52-114 g/L of erythritol. The productivity was 0.58-1.04 g/L/h, and yield was 0.26-0.57 g/g; the final biomasses yield ranged 17-41 g/L. This is the first report describing erythritol production via industrial raw molasses and glycerol by Y. lipolytica. This work uses genetically modified strains of Y. lipolytica as tool for the direct conversion of affordable raw industrial molasses and glycerol into the value-added erythritol product.

Polyol production from waste materials by genetically modified Yarrowia lipolytica

Sugar alcohols (polyols) are sweeteners with many industrial applications. In this study, a fermentation process of polyol production based on waste substrates - raw industrial molasses and crude glycerol - was tested. The yeast strain Yarrowia lipolytica Wratislavia K1 was genetically modified by overexpression of the Saccharomyces cerevisiae SUC2 gene and overexpression of the native GUT1 gene. This process allowed for sucrose utilization and rapid glycerol assimilation by the engineered strain. In this study, the obtained strain AIB pAD-UTGut1 produced 100.65±3.75g/l of polyols, with productivity of 1.09±0.9g/lh and yield of 0.67±0.2g/g. This is the first study describing efficient polyol production by the modified Y. lipolytica strain from industrial raw molasses and crude glycerol. By process optimization, we established conditions for abundant polyol synthesis from low-value substrates.

Recent advances in biological production of erythritol

Abstract Erythritol is a natural sweetener commonly used in the food and pharmaceutical industries. Produced by microorganisms as an osmoprotectant, it is an ideal sucrose substitute for diabetics or overweight persons due to its almost zero calorie content. Currently, erythritol is produced on an industrial scale through the fermentation of sugars by some yeasts, such as Moniliella sp. However, the popularity of erythritol as a sweetener is still small because of its high retail price. This creates an opportunity for further process improvement. Recent years have brought the rapid development of erythritol biosynthesis methods from the low-cost substrates, and a better understanding of the metabolic pathways leading to erythritol synthesis. The yeast Yarrowia lipolytica emerges as an organism effectively producing erythritol from pure or crude glycerol. Moreover, novel erythritol producing organisms and substrates may be taken into considerations due to metabolic engineering. This review focuses on the modification of erythritol production to use low-cost substrates and metabolic engineering of the microorganisms in order to improve yield and productivity.

Synthesis, Surface and Antimicrobial Activity of Piperidine-Based Sulfobetaines

A new method for the preparation of new heterocyclic amine surfactants based on sulfobetaines is proposed. Interfacial activities of the surfactants obtained in aqueous solution were studied by surface tension measurements. The critical micelle concentration, surface excess concentration, minimum area per surfactant molecule, and standard Gibbs energy of adsorption were determined. The adsorption properties of these compounds depend significantly on the alkyl chain length. Alkyl chain length also affects biological properties of the new surfactants, determining the minimum inhibitory concentration and size of inhibited growth zone. The compounds have high antimicrobial activity.

A Role of a Newly Identified Isomerase From Yarrowia lipolytica in Erythritol Catabolism

Erythritol is a natural sweetener produced by microorganisms as an osmoprotectant. It belongs to the group of polyols and it can be utilized by the oleaginous yeast Yarrowia lipolytica. Despite the recent identification of the transcription factor of erythritol utilization (EUF1), the metabolic pathway of erythritol catabolism remains unknown. In this study we identified a new gene, YALI0F01628g, involved in erythritol assimilation. In silico analysis showed that YALI0F01628g is a putative isomerase and it is localized in the same region as EUF1. qRT-PCR analysis of Y. lipolytica showed a significant increase in YALI0F01628g expression during growth on erythritol and after overexpression of EUF1. Moreover, the deletion strain ΔF01628 showed significantly impaired erythritol assimilation, whereas synthesis of erythritol remained unchanged. The results showed that YALI0F1628g is involved in erythritol assimilation; thus we named the gene EYI1. Moreover, we suggest the metabolic pathway of erythritol assimilation in yeast Y. lipolytica.

Aseptic production of citric and isocitric acid from crude glycerol by genetically modified Yarrowia lipolytica

The unconventional yeast Yarrowia lipolytica is known for its capacity to produce citric or isocitric acid from glycerol. In this study a reduction of production cost was achieved by using cheap crude glycerol and conducting the production at pH 3 to prevent bacterial contamination. In this study a Y. lipolytica strain overexpressing Gut1 and Gut2 was used. For the modified strain, crude glycerol proved to be an excellent substrate for production of citric/isocitric acids in aseptic conditions, as the final concentration of these compounds reached 75.9 ± 1.8 g L-1 after 7 days of batch production. Interestingly, the concentration of isocitric acid was 42.5 ± 2.4 g L-1, which is one of the highest concentrations of isocitric acid obtained from a waste substrate. In summary, these data show that organic acids can be efficiently produced by the yeast Y. lipolytica from crude glycerol without any prior purification in aseptic conditions.