Biotechnological potential of red yeast isolated from birch forests in Poland.

OBJECTIVES: This study aimed to isolate red yeast from sap, bark and slime exudates collected from Polish birch forests and then assessment of their biotechnological potential. RESULTS: 24 strains of red yeast were isolated from the bark, sap and spring slime fluxes of birch (Betula pendula). Strains belonging to Rhodotorula mucilaginosa (6), Rhodosporidiobolus colostri (4), Cystrofilobasidium capitaum (3), Phaffia rhodozyma (3) and Cystobasidium psychroaquaticum (3) were dominant. The highest efficiency of carotenoid biosynthesis (5.04 mg L-1) was obtained by R. mucilaginosa CMIFS 004, while lipids were most efficiently produced by two strains of P. rhodozyma (5.40 and 5.33 g L-1). The highest amount of exopolysaccharides (3.75 g L-1) was produced by the R. glutinis CMIFS 103. Eleven strains showed lipolytic activity, nine amylolytic activity, and only two proteolytic activity. The presence of biosurfactants was not found. The growth of most species of pathogenic moulds was best inhibited by Rhodotorula yeasts. CONCLUSION: Silver birch is a good natural source for the isolation of new strains of red yeast with wide biotechnological potential.

Physiological and genetic regulation of anhydrobiosis in yeast cells.

Anhydrobiosis is a state of living organisms during which their metabolism is reversibly delayed or suspended due to a high degree of dehydration. Yeast cells, which are widely used in the food industry, may be induced into this state. The degree of viability of yeast cells undergoing the drying process also depends on rehydration. In an attempt to explain the essence of the state of anhydrobiosis and clarify the mechanisms responsible for its course, scientists have described various cellular compounds and structures that are responsible for it. The structures discussed in this work include the cell wall and plasma membrane, vacuoles, mitochondria, and lysosomes, among others, while the most important compounds include trehalose, glycogen, glutathione, and lipid droplets. Various proteins (Stf2p; Sip18p; Hsp12p and Hsp70p) and genes (STF2; Nsip18; TRX2; TPS1 and TPS2) are also responsible for the process of anhydrobiosis. Each factor has a specific function and is irreplaceable, detailed information is presented in this overview.

Comparison of the Chemical Composition and Antioxidant Properties of Propolis from Urban Apiaries.

Bee products from urban apiaries are increasingly used. They are mainly used to promote local apiaries and cities in which they are located. The aim of the study was to compare the chemical composition and antioxidant activity of propolis from 6 Polish apiaries located in cities (Legionowo, Torun, Cracow, Warsaw, Katowice, Lodz). The chemical composition was analyzed using liquid chromatography (HPLC-DAD) and the analysis of antioxidant activity by scavenging free radicals (ABTS and DPPH) and FRAP. The obtained results showed the presence of 24 phenolic compounds in propolis extracts. The tested samples showed differentiation in terms of the content of individual chemical components, however, cinnamic acid and its derivatives were dominant. High antioxidant activity of the tested extracts was demonstrated (ABTS was in the range of 16.80-51.53 mg Te/mL, DPPH was in the range of 7.54-22.13 mg Te/mL, while FRAP reduction was in the range of 10.93-29.55 mg Te/mL). The obtained results compared with literature data on propolis from agricultural areas allow to conclude that propolis samples from both Poland types of areas are similar and can be classified as poplar propolis.

Recent advances and opportunities related to the use of bee products in food processing.

Nowadays, natural foods that can provide positive health effects are gaining more and more popularity. Bees and the products they produce are our common natural heritage that should be developed. In the article, we presented the characteristics of bee products and their use in industry. We described the development and importance of beekeeping in the modern world. Due to their high nutritional value and therapeutic properties, bee products are of great interest and their consumption is constantly growing. The basis for the use of bee products in human nutrition is their properties and unique chemical composition. The conducted research and opinions confirm the beneficial effect of bee products on health. The current consumer awareness of the positive impact of food having a pro-health effect on health and well-being affects the increase in interest and demand for this type of food among various social groups. Enriching the daily diet with bee products may support the functioning of the organism. New technologies have appeared on the market to improve the process of obtaining bee products. The use of bee products plays a large role in many industries; moreover, the consumption of bee products and promotion of their medicinal properties are very important in shaping proper eating habits.

Magnesium Binding by Cyberlindnera jadinii Yeast in Media from Potato Wastewater and Glycerol.

The aim of this study was to determine the magnesium-binding capacity of Cyberlindnera jadinii yeast in media prepared from potato wastewater and glycerol (after biodiesel production), supplemented with magnesium salt. The research was carried out in two stages. In the first, the ability to binding magnesium by yeast in media supplemented with various doses of this element was tested. In the second stage, after selecting the appropriate dose of magnesium, the culture was carried out in a bioreactor. The composition of the yeast biomass was also analysed in terms of lipids and protein content and amino acid composition. Studies have shown that this type of medium can be used as a culture medium for the growth of C. jadinii yeast. In the first stage of the study, the most magnesium (8.97 mg/gd.m.) was bound by yeast cells after 48 h of cultivation in a medium supplemented with the addition of magnesium at a dose of 2 g/L. In the second stage of the research, the highest magnesium content in the biomass (7.9 mg/gd.m.) was noted after 24 h of cultivation in the same medium. The lipid and protein contents in the biomass obtained after 24 h of cultivation in the bioreactor were 6.35 and 43.73%, respectively. The main fatty acids present in the yeast lipids were oleic acid (59.4%) and linoleic acid (8.6%). Analysis of the amino acid profile of the proteins showed the highest proportions were glutamic acid (13.7%) and aspartic acid (11%).

Diversity of Red Yeasts in Various Regions and Environments of Poland and Biotechnological Potential of the Isolated Strains.

Due to the growing demand for natural carotenoids, researchers have been searching for strains that are capable of efficient synthesis of these compounds. This study tested 114 red yeast strains collected from various natural environments and food specimens in Poland. The strains were isolated by their ability to produce red or yellow pigments in rich nutrient media. According to potential industrial significance of the carotenoids, both their total production and share of individual carotenoids (ß-carotene, γ-carotene, torulene, and torularhodin) were analyzed. The total content of carotenoid pigments in the yeast dry matter ranged from 13.88 to 406.50 µg/g, and the percentages of individual carotenoids highly varied among the strains. Most of the yeast isolates synthesized torulene at the highest amount. Among the studied strains, isolates with a total carotenoid content in biomass greater than 200 µg/g and those containing more than 60% torularhodin were selected for identification (48 strains). The identified strains belonged to six genera: Rhodotorula, Sporidiobolus, Sporobolomyces, Buckleyzyma, Cystofilobasidium, and Erythrobasidium. The largest number of isolates belonged to Rhodotorula babjevae (18), Rhodotorula mucilaginosa (7), Sporidiobolus pararoseus (4), and Rhodotorula glutinis (4).

Effect of selenium and methods of protein extraction on the proteomic profile of Saccharomyces yeast.

Selenium may influence the biosynthesis of individual proteins in the yeast cell cytosol. In this study, we used two-dimensional (2D) electrophoresis to identify proteins that are differentially expressed by the enrichment of selenium in Saccharomyces cerevisiae yeast cells. We chose eight protein fractions for further proteomic analysis. A detailed analysis was performed using the Ultraflextreme matrix-assisted laser desorption/ionisation time-of-flight/time-of-flight mass spectrometer, which enables fast and accurate measurement of the molecular weight of the analysed proteins. This study, for the first time, provides evidence that selenium-enriched yeast contains higher levels of mitochondria malate dehydrogenase, adenosine-5'-triphosphate (ATP)-dependent RNA helicase dbp3, and tryptophan dimethylallyltransferase, and alanyl-tRNA editing protein AlaX than yeast without the addition of selenium. It should be emphasised that the proteomic variability obtained reflects the high biological and complexity of yeast metabolism under control and selenium-enriched conditions and can be properly used in the future as a model for further research aimed at determining the expression of appropriate metabolic genes.

Characteristics of the Proteolytic Enzymes Produced by Lactic Acid Bacteria.

Over the past several decades, we have observed a very rapid development in the biotechnological use of lactic acid bacteria (LAB) in various branches of the food industry. All such areas of activity of these bacteria are very important and promise enormous economic and industrial successes. LAB are a numerous group of microorganisms that have the ability to ferment sugars into lactic acid and to produce proteolytic enzymes. LAB proteolytic enzymes play an important role in supplying cells with the nitrogen compounds necessary for their growth. Their nutritional requirements in this regard are very high. Lactic acid bacteria require many free amino acids to grow. The available amount of such compounds in the natural environment is usually small, hence the main function of these enzymes is the hydrolysis of proteins to components absorbed by bacterial cells. Enzymes are synthesized inside bacterial cells and are mostly secreted outside the cell. This type of proteinase remains linked to the cell wall structure by covalent bonds. Thanks to advances in enzymology, it is possible to obtain and design new enzymes and their preparations that can be widely used in various biotechnological processes. This article characterizes the proteolytic activity, describes LAB nitrogen metabolism and details the characteristics of the peptide transport system. Potential applications of proteolytic enzymes in many industries are also presented, including the food industry.

Comparison of simple and rapid cell wall disruption methods for improving lipid extraction from yeast cells.

The present study examined the effect of six disruption methods of the cell wall (acid hydrolysis, ultrasonication, osmotic shock, pasteurization, homogenization with zirconia balls, and freezing/defrosting) on the efficiency of lipid extraction from yeast cells and the composition of fatty acids. Acid hydrolysis and sonication led to a significant increase in lipid extraction from Cyberlindnera jadinii ATCC 9950 and Rhodotorula glutinis LOCKR13 yeast cells. The amount of lipids extracted in these conditions increased for C. jadinii from 12.46 (biomass not subjected to any pretreatment) to 20.37 and 19.53 g/100 gd.w. after the application of acid hydrolysis and sonication, respectively, and for R. glutinis strain from 13.95 to 21.20 and 17.22 g/100 gd.w., respectively, for the same methods. Initial sonication of biomass led to a significant reduction in the percentage of unsaturated fatty acids. The largest differences in fatty acid composition were found for the sample homogenized with zirconium balls. This process resulted in the degradation of both oleic acid and linolenic acid. The obtained results revealed that the method that significantly increases lipid extraction and does not change the composition of fatty acids is acid hydrolysis with hydrochloric acid. In addition, it is easy, cheap, does not require specialized equipment, and therefore can be implemented in any laboratory.

The aspects of microbial biomass use in the utilization of selected waste from the agro-food industry.

Cellular biomass of microorganisms can be effectively used in the treatment of waste from various branches of the agro-food industry. Urbanization processes and economic development, which have been intensifying in recent decades, lead to the degradation of the natural environment. In the first half of the 20th century, problems related to waste management were not as serious and challenging as they are today. The present situation forces the use of modern technologies and the creation of innovative solutions for environmental protection. Waste of industrial origin are difficult to recycle and require a high financial outlay, while the organic waste of animal and plant origins, such as potato wastewater, whey, lignin, and cellulose, is dominant. In this article, we describe the possibilities of using microorganisms for the utilization of various waste products. A solution to reduce the costs of waste disposal is the use of yeast biomass. Management of waste products using yeast biomass has made it possible to generate new metabolites, such as ß-glucans, vitamins, carotenoids, and enzymes, which have a wide range of industrial applications. Exploration and discovery of new areas of applications of yeast, fungal, and bacteria cells can lead to an increase in their effective use in many fields of biotechnology.

Effect of exogenous stress factors on the biosynthesis of carotenoids and lipids by Rhodotorula yeast strains in media containing agro-industrial waste.

In this study, we aimed to determine the effect of exogenous stress factors (sodium chloride as osmotic stressor, hydrogen peroxide as an inducer of oxidative stress, white light irradiation, and low temperature) on the biosynthesis of carotenoids and lipids by red yeast (Rhodotorula glutinis, R. mucilaginosa, and R. gracilis) during cultivation in media containing potato wastewater and glycerol. According to our results, the yeast were able to grow and biosynthesize lipids and carotenoids in the presence of the applied stress factors. Low temperature caused an increase in the biosynthesis of intracellular lipids and carotenoids. R. gracilis synthesized lipids (21.1 g/100 gd.w.) and carotenoids (360.4 µg/gd.w.) in greater quantities than that of other strains. Under these conditions, there was also an increase in the content of unsaturated fatty acids, especially linoleic and linolenic acids. The highest percentage of polyunsaturated fatty acid (PUFA) (30.4%) was synthesized by the R. gracilis yeast after cultivation at 20°C. Their quantity was 2.5-fold greater than that of the biomass grown in control conditions. The contribution of individual carotenoid fractions depended both on the yeast strain and the culture conditions. Induction of osmotic stress and low temperature intensified the biosynthesis of ß-carotene (up to 73.9% of the total carotenoid content). In oxidative stress conditions, yeast synthesized torulene (up to 82.2%) more efficiently than under other conditions, whereas white light irradiation increased the production of torularhodin (up to 20.0%).

Simultaneous Production of Lipids and Carotenoids by the Red Yeast Rhodotorula from Waste Glycerol Fraction and Potato Wastewater.

AbstractThe objective of this study was to determine the possibility of simultaneous biosynthesis of lipids and carotenoids by the Rhodotorula yeast strains in media with waste glycerol and deproteinized potato wastewater and to determine the level of pollution reduction by media. On the basis of results obtained during the yeast microcultures in the Bioscreen C system, it was found that potato wastewater and glycerol can be used as components of media for Rhodotorula glutinis, Rhodotorula mucilaginosa, and Rhodotorula gracilis yeast strains. The amount of glycerol added to media higher than 10% significantly decreased the growth rate of yeast. The results of yeast culture in the laboratory shaker flasks showed a possibility of simultaneous production of lipids and carotenoids by R. glutinis, R. mucilaginosa, and R. gracilis yeast strains during cultivation in media containing only waste glycerol and deproteinized potato wastewater. A higher intracellular lipid content (approximately 15 g/100 gd.w.) was obtained for R. mucilaginosa and R. gracilis yeast biomass after cultivation in experimental media with waste glycerol and potato wastewater. In conclusion, the yeast grown in media with potato wastewater supplemented with 3% or 5% glycerol synthesized carotenoids, and their content in biomass did not exceed 230 µg/gd.w.

Effect of selenium on growth and antioxidative system of yeast cells.

Selenium exhibits health-promoting properties in humans and animals. Therefore, the development of selenium-enriched dietary supplements has been growing worldwide. However, it may also exhibit toxicity at higher concentrations, causing increased oxidative stress. Different species of yeasts may exhibit different tolerances toward selenium. Therefore, in this study, we aimed to determine the effect of selenium on growth and on the antioxidative system in Candida utilis ATCC 9950 and Saccharomyces cerevisiae ATCC MYA-2200 yeast cells. The results of this study have demonstrated that high doses of selenium causes oxidative stress in yeasts, thereby increasing the process of lipid peroxidation. In addition, we obtained an increased level of GSSG from aqueous solutions of yeast biomass grown with selenium supplementation (40-60 mg/L). Increased levels of selenium in aqueous solutions resulted in an increase in the activity of antioxidant enzymes, including glutathione peroxidase and glutathione reductase. These results should encourage future research on the possibility of a thorough understanding of antioxidant system functioning in yeast cells.

Effect of Selenium on Lipid and Amino Acid Metabolism in Yeast Cells.

This article discusses the effect of selenium in aqueous solutions on aspects of lipid and amino acid metabolism in the cell biomass of Saccharomyces cerevisiae MYA-2200 and Candida utilis ATCC 9950 yeasts. The yeast biomass was obtained by using waste products (potato wastewater and glycerol). Selenium, at a dose of 20 mg/L of aqueous solution, affected the differentiation of cellular morphology. Yeast enriched with selenium was characterized by a large functional diversity in terms of protein and amino acid content. The protein content in the biomass of S. cerevisiae enriched with selenium (42.6%) decreased slightly as compared to that in the control sample without additional selenium supplementation (48.4%). Moreover, yeasts of both strains enriched with selenium contained a large amount of glutamic acid, aspartic acid, lysine, and leucine. Analysis of fatty acid profiles in S. cerevisiae yeast supplemented with selenium showed an increase in the unsaturated fatty acid content (e.g., C18:1). The presence of margaric acid (C17:0) and hexadecanoic acid (C17:1) was found in the C. utilis biomass enriched with selenium, in contrast to that of S. cerevisiae. These results indicate that selenium may induce lipid peroxidation, which consequently affects the loss of integrity of the cytoplasmic membrane. Yeast enriched with selenium with optimal amino acid and lipid composition can be used to prepare a novel formula of dietary supplements, which can be applied directly to various diets for both humans and animals.

Correction to: Effect of Selenium on Lipid and Amino Acid Metabolism in Yeast Cells.

The authors forgot to include the following information in Materials and Methods.

Torulene and torularhodin: "new" fungal carotenoids for industry?

Torulene and torularhodin represent the group of carotenoids and are synthesized by yeasts and fungi. The most important producers of these two compounds include yeasts of Rhodotorula and Sporobolomyces genera. The first reports confirming the presence of torulene and torularhodin in the cells of microorganisms date to the 1930s and 1940s; however, only in the past few years, the number of works describing the properties of these compounds increased. These compounds have strong anti-oxidative and anti-microbial properties, and thus may be successfully used as food, feedstock, and cosmetics additives. In addition, tests performed on rats and mice showed that both torulene and torularhodin have anti-cancerous properties. In order to commercialize the production of these two carotenoids, it is necessary to obtain highly efficient yeast strains, for example, via mutagenization and optimization of cultivation conditions. Further studies on the activity of torulene and torularhodin on the human body are also needed.

Effect of initial pH of medium with potato wastewater and glycerol on protein, lipid and carotenoid biosynthesis by Rhodotorula glutinis

Background: Rhodotorula glutinis is capable of synthesizing numerous valuable metabolites with extensive potential industrial usage. This paper reports the effect of initial culture medium pH on growth and protein, lipid, and carotenoid biosynthesis by R. glutinis. Results: The highest biomass yield was obtained in media with pH 4.0­7.0, and the value after 72 h was 17.2­19.4 gd.w./L. An initial pH of the medium in the range of 4.0­7.0 has no significant effect on the protein (38.5­41.3 g/100 gd.w.), lipid (10.2­12.7 g/100 gd.w.), or carotenoid (191.7­202.9 µg/gd.w.) content in the biomass or on the profile of synthesized fatty acids and carotenoids. The whole pool of fatty acids was dominated by oleic (48.1­53.4%), linoleic (21.4­25.1%), and palmitic acids (13.0­15.8%). In these conditions, the yeast mainly synthesized torulene (43.5­47.7%) and ß-carotene (34.7­38.6%), whereas the contribution of torularhodin was only 12.1­16.8%. Cultivation in medium with initial pH 3.0 resulted in a reduction in growth (13.0 gd.w./L) and total carotenoid (115.8 µg/gd.w.), linoleic acid (11.5%), and torularhodin (4.5%) biosynthesis. Conclusion: The different values of initial pH of the culture medium with glycerol and deproteinized potato wastewater had a significant effect on the growth and protein, lipid, and carotenoid biosynthesis by R. glutinis.

Rhodotorula glutinis-potential source of lipids, carotenoids, and enzymes for use in industries.

Rhodotorula glutinis is capable of synthesizing numerous valuable compounds with a wide industrial usage. Biomass of this yeast constitutes sources of microbiological oils, and the whole pool of fatty acids is dominated by oleic, linoleic, and palmitic acid. Due to its composition, the lipids may be useful as a source for the production of the so-called third-generation biodiesel. These yeasts are also capable of synthesizing carotenoids such as ß-carotene, torulene, and torularhodin. Due to their health-promoting characteristics, carotenoids are commonly used in the cosmetic, pharmaceutical, and food industries. They are also used as additives in fodders for livestock, fish, and crustaceans. A significant characteristic of R. glutinis is its capability to produce numerous enzymes, in particular, phenylalanine ammonia lyase (PAL). This enzyme is used in the food industry in the production of L-phenylalanine that constitutes the substrate for the synthesis of aspartame-a sweetener commonly used in the food industry.