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.

Reprocessing of side-streams towards obtaining valuable bacterial metabolites.

Every year, all over the world, the industry generates huge amounts of residues. Side-streams are most often used as feed, landfilled, incinerated, or discharged into sewage. These disposal methods are far from perfect. Taking into account the composition of the side-streams, it seems that they should be used as raw materials for further processing, in accordance with the zero-waste policy and sustainable development. The article describes the latest achievements in biotechnology in the context of bacterial reprocessing of residues with the simultaneous acquisition of their metabolites. The article focuses on four metabolites - bacterial cellulose, propionic acid, vitamin B12 and PHAs. Taking into account global trends (e.g. food, packaging, medicine), it seems that in the near future there will be a sharp increase in demand for this type of compounds. In order for their production to be profitable and commercialised, cheap methods of its obtaining must be developed. The article, in addition to obtaining these bacterial metabolites from side-streams, also discusses e.g. factors affecting their production, metabolic pathways and potential and current applications. The presented chapters provide a complete overview of the current knowledge on above metabolites, which can be helpful for the academic and scientific communities and the several industries. KEY POINTS: • The industry generates millions of tons of organic side-streams each year. • Generated residues burden the natural environment. • A good and cost-effective method of side-streams management seems to be biotechnology - reprocessing with the use of bacteria. • Biotechnological disposal of side-streams gives the opportunity to obtain valuable compounds in cheaper ways: BC, PA, vitmain B12, PHAs.

In silico and in vitro analysis of a rational mutation in gIII signal peptide and its effects on periplasmic expression of rhGH in E. coli.

The secretion efficiency of a heterologous protein in E. coli is mainly dictated by the N-terminal signal peptide fused to the desired protein. In this study, we aimed to select and introduce mutations into the - 1, - 2 and - 3 positions of the gIII signal peptide (originated from filamentous phage fd Gene III) fused to the N-terminus of the human growth hormone (hGH), and study its effect on the secretion efficiency of the recombinant hGH into the periplasmic space of E. coli Top10. Bioinformatics software such as SignalP-5.0 and PrediSi were employed to predict the effects of the mutations on the secretion efficiency of the recombinant hGH. Site-directed mutagenesis was applied to introduce the desired mutations into the C-terminus of the gIII signal peptide. The periplasmic expression and the secretion efficiency of the recombinant hGH using the native and mutant gIII signal peptides were compared in E. coli Top10 under the control of araBAD promoter. Our results from bioinformatics analysis indicated that the mutant gIII signal peptide was more potent than the native one for secretion of the recombinant hGH in E. coli. While our experimental results revealed that the mutation had no effect on hGH secretion. This result points to the importance of experimental validation of bioinformatics predictions.

Use of apple pomace, glycerine, and potato wastewater for the production of propionic acid and vitamin B12.

Propionic acid bacteria (PAB) are a source of valuable metabolites, including propionic acid and vitamin B12. Propionic acid, a food preservative, is synthesized from petroleum refining by-products, giving rise to ecological concerns. Due to changing food trends, the demand for vitamin B12 has been expected to increase in the future. Therefore, it is necessary to look for new, alternative methods of obtaining these compounds. This study was conducted with an aim of optimizing the production of PAB metabolites using only residues (apple pomace, waste glycerine, and potato wastewater), without any enzymatic or chemical pretreatment and enrichment. Media consisting of one, two, or three industrial side-streams were used for the production of PAB metabolites. The highest production of propionic acid was observed in the medium containing all three residues (8.15 g/L, yield: 0.48 g/g). In the same medium, the highest production of acetic acid was found - 2.31 g/L (0.13 g/g). The presence of waste glycerine in the media had a positive effect on the efficiency of propionic acid production and P/A ratio. The concentration of vitamin B12 obtained in the wet biomass of Propionibacterium freudenreichii DSM 20271 ranged from 90 to 290 µg/100 g. The highest production of cobalamin was achieved in potato wastewater and apple pomace, which may be a source of the precursors of vitamin B12 - cobalt and riboflavin. The results obtained show both propionic acid and vitamin B12 can be produced in a more sustainable manner through the fermentation of residues which are often not properly managed. KEY POINTS: • The tested strain has been showed metabolic activity in the analyzed industrial side-streams (apple pomace, waste glycerine, potato wastewater). • All the side-streams were relevant for the production of propinic acid. • The addition of waste glycerine increases the propionic acid production efficiency and P/A ratio. • B12 was produced the most in the media containing potato wastewater and apple pomace as dominant ingredients.

Astounding Health Benefits of Jamun (Syzygium cumini) toward Metabolic Syndrome.

Syzygium cumini, also called Jamun, or black plum, is an excellent source of bioactive components such as flavonoids, polyphenols, antioxidants, iron, and vitamin C. The Jamun tree is a tropical evergreen blooming plant and is an important medicinal plant from the Myrtaceae family that has been used for a long time in Indian and other traditional medicines across the world. Jamun is mainly cultivated in Asian countries such as Pakistan, India, Sri Lanka, and Bangladesh. Since ancient medicine, it has been utilized to treat a variety of diseases and physiological conditions. Currently, it is mostly used as a medication to treat various metabolic issues, including diabetes, hyperlipidemia, hypertension, obesity, etc. Therefore, Jamun could serve a beneficial role against metabolic syndrome (MS). In this work, the latest available scientific literature on Jamun was collected and the clinical trials investigating its effect on diabetes, hypertension, obesity, and hyperlipidemia were analyzed to find out how Jamun could improve the symptoms and biomarkers of MS. Overall, the results of this study found a significant association of Jamun with the prevention and treatment of these biomarkers of MS. In many studies, Jamun showed pharmacological modifications not only in MS but in many other diseases as well. Currently, its utilization as a folk medicine for the treatment of patients with MS is widely acknowledged. Hence, the findings of a large number of clinical studies confirmed the ameliorating effects of Jamun against MS due to its antioxidation, antidiabetic, anti-inflammation anticarcinogenic, and hyperlipidemic effects. More research is still needed to determine and identify the Jamun compounds and to elucidate their mechanisms of action that are responsible for these astounding bioactive properties and health benefits.

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.

An Intricate Review on Nutritional and Analytical Profiling of Coconut, Flaxseed, Olive, and Sunflower Oil Blends.

Vegetable oils (VOs), being our major dietary fat source, play a vital role in nourishment. Different VOs have highly contrasting fatty acid (FA) profiles and hence possess varying levels of health protectiveness. Consumption of a single VO cannot meet the recommended allowances of various FA either from saturated FA (SFA), monounsaturated FA (MUFA), polyunsaturated FA (PUFA), Ω-3 PUFAs, and medium-chain triglycerides (MCTs). Coconut oil (CO), flaxseed oil (FO), olive oil (OO), and sunflower oil (SFO) are among the top listed contrast VOs that are highly appreciated based on their rich contents of SFAs, Ω-3 PUFAs, MUFAs, and Ω-6 PUFA, respectively. Besides being protective against various disease biomarkers, these contrasting VOs are still inappropriate when consumed alone in 100% of daily fat recommendations. This review compiles the available data on blending of such contrasting VOs into single tailored blended oil (BO) with suitable FA composition to meet the recommended levels of SFA, MUFA, PUFA, MCTs, and Ω-3 to Ω-6 PUFA ratios which could ultimately serve as a cost-effective dietary intervention towards the health protectiveness and improvement of the whole population in general. The blending of any two or more VOs from CO, FO, OO, and SFO in the form of binary, ternary, or another type of blending was found to be very conclusive towards balancing FA composition; enhancing physiochemical and stability properties; and promising the therapeutic protectiveness of the resultant BOs.

Use of Propionibacterium freudenreichii T82 Strain for Effective Biosynthesis of Propionic Acid and Trehalose in a Medium with Apple Pomace Extract and Potato Wastewater.

Propionic acid bacteria are the source of many metabolites, e.g., propionic acid and trehalose. Compared to microbiological synthesis, the production of these metabolites by petrochemical means or enzymatic conversion is more profitable. The components of microbiological media account for a large part of the costs associated with propionic fermentation, due to the high nutritional requirements of Propionibacterium. This problem can be overcome by formulating a medium based on the by-products of technological processes, which can act as nutritional sources and at the same time replace expensive laboratory preparations (e.g., peptone and yeast extract). The metabolic activity of P. freudenreichii was investigated in two different breeding environments: in a medium containing peptone, yeast extract, and biotin, and in a waste-based medium consisting of only apple pomace and potato wastewater. The highest production of propionic acid amounting to 14.54 g/L was obtained in the medium containing apple pomace and pure laboratory supplements with a yield of 0.44 g/g. Importantly, the acid production parameters in the waste medium reached almost the same level (12.71 g/L, 0.42 g/g) as the medium containing pure supplements. Acetic acid synthesis was more efficient in the waste medium; it was also characterized by a higher level of accumulated trehalose (59.8 mg/g d.s.). Thus, the obtained results show that P. freudenreichii bacteria exhibited relatively high metabolic activity in an environment with apple pomace used as a carbon source and potato wastewater used as a nitrogen source. This method of propioniate production could be cheaper and more sustainable than the chemical manner.

Phytochemistry, Food Application, and Therapeutic Potential of the Medicinal Plant (Withania coagulans): A Review.

Herbal plants have been utilized to treat and cure various health-related problems since ancient times. The use of Ayurvedic medicine is very significant because of its least reported side effects and host of advantages. Withania coagulans (Family; Solanaceae), a valuable medicinal plant, has been used to cure abnormal cell growth, wasting disorders, neural as well as physical problems, diabetes mellitus, insomnia, acute and chronic hepatic ailments. This review provides critical insight regarding the phytochemistry, biological activities, and pharmacognostic properties of W. coagulans. It has been known to possess diuretic, anti-inflammatory, anti-bacterial, anti-fungal, cardio-protective, hepato-protective, hypoglycemic, anti-oxidative, and anti-mutagenic properties owing to the existence of withanolides, an active compound present in it. Apart from withanolides, W. coagulans also contains many phytochemicals such as flavonoids, tannins, and ß-sterols. Several studies indicate that various parts of W. coagulans and their active constituents have numerous pharmacological and therapeutic properties and thus can be considered as a new drug therapy against multiple diseases.

Ultrasound-Assisted Extraction of Carotenoids from Carrot Pomace and Their Optimization through Response Surface Methodology.

Ultrasound-assisted extraction (UAE) was used to extract carotenoids from the carrot pomace. To investigate the effect of independent variables on the UAE, the response surface methodology (RSM) with central-composite design (CCD) was employed. The study was conducted with three independent variables including extraction time (min), temperature (°C), and ethanol concentration (%). The results showed that the optimal conditions for UAE were achieved with an extraction time of 17 min, temperature of 32 °C, and ethanol concentration of 51% of total carotenoids (31.82 ± 0.55); extraction time of 16 min, temperature of 29 °C, and ethanol concentration of 59% for a combination of ß-carotene (14.89 ± 0.40), lutein (5.77 ± 0.19), and lycopene (2.65 ± 0.12). The non-significant (p > 0.05) correlation under optimal extraction conditions between predicted and experimental values suggested that UAE is the more productive process than conventional techniques for the extraction of carotenoids from the carrot pomace.

A Critical Review on Pulsed Electric Field: A Novel Technology for the Extraction of Phytoconstituents.

Different parts of a plant (seeds, fruits, flower, leaves, stem, and roots) contain numerous biologically active compounds called "phytoconstituents" that consist of phenolics, minerals, amino acids, and vitamins. The conventional techniques applied to extract these phytoconstituents have several drawbacks including poor performance, low yields, more solvent use, long processing time, and thermally degrading by-products. In contrast, modern and advanced extraction nonthermal technologies such as pulsed electric field (PEF) assist in easier and efficient identification, characterization, and analysis of bioactive ingredients. Other advantages of PEF include cost-efficacy, less time, and solvent consumption with improved yields. This review covers the applications of PEF to obtain bioactive components, essential oils, proteins, pectin, and other important materials from various parts of the plant. Numerous studies compiled in the current evaluation concluded PEF as the best solution to extract phytoconstituents used in the food and pharmaceutical industries. PEF-assisted extraction leads to a higher yield, utilizes less solvents and energy, and it saves a lot of time compared to traditional extraction methods. PEF extraction design should be safe and efficient enough to prevent the degradation of phytoconstituents and oils.

Metabolic Response of the Yeast Candida utilis During Enrichment in Selenium.

Selenium (Se) was found to inhibit the growth of the yeast Candida utilis ATCC 9950. Cells cultured in 30 mg selenite/L supplemented medium could bind 1368 µg Se/g of dry weight in their structures. Increased accumulation of trehalose and glycogen was observed, which indicated cell response to stress conditions. The activity of antioxidative enzymes (glutathione peroxidase, glutathione reductase, thioredoxin reductase, and glutathione S-transferase) was significantly higher than that of the control without Se addition. Most Se was bound to water-insoluble protein fraction; in addition, the yeast produced 20-30 nm Se nanoparticles (SeNPs). Part of Se was metabolized to selenomethionine (10%) and selenocysteine (20%). The HPLC-ESI-Orbitrap MS analysis showed the presence of five Se compounds combined with glutathione in the yeast. The obtained results form the basis for further research on the mechanisms of Se metabolism in yeast cells.

Biological Activity of Some Aromatic Plants and Their Metabolites, with an Emphasis on Health-Promoting Properties.

The biological activities of four aromatic plants, namely frankincense, myrrh, ginger, and turmeric, were reviewed in the current study. The volatile fraction (essential oil) as well as the nonvolatile fraction of these four plants showed different promising biological activities that are displayed in detail. These activities can include protection from and/or alleviation of some ailment, which is supported with different proposed mechanisms of action. This review aimed to finally help researchers to get a handle on the importance of considering these selected aromatic plants, which have not been thoroughly reviewed before, as a potential adjuvant to classical synthetic drugs to enhance their efficiency. Moreover, the results elicited in this review encourage the consumption of these medicinal plants as an integrated part of the diet to boost the body's overall health based on scientific evidence.

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.

Selenium⁻Fascinating Microelement, Properties and Sources in Food.

Selenium is a micronutrient that is essential for the proper functioning of all organisms. Studies on the functions of selenium are rapidly developing. This element is a cofactor of many enzymes, for example, glutathione peroxidase or thioredoxin reductase. Insufficient supplementation of this element results in the increased risk of developing many chronic degenerative diseases. Selenium is important for the protection against oxidative stress, demonstrating the highest activity as a free radical scavenger and anti-cancer agent. In food, it is present in organic forms, as exemplified by selenomethionine and selenocysteine. In dietary supplementation, the inorganic forms of selenium (selenite and selenate) are used. Organic compounds are more easily absorbed by human organisms in comparison with inorganic compounds. Currently, selenium is considered an essential trace element of fundamental importance for human health. Extreme selenium deficiencies are widespread among people all over the world. Therefore, it is essential to supplement the deficiency of this micronutrient with selenium-enriched food or yeast cell biomass in the diet.

Optimization of propionic acid production in apple pomace extract with Propionibacterium freudenreichii.

Sequential optimization of propionate production using apple pomace was studied. All experiments were performed in a static flask in anaerobic conditions. Effect of apple pomace as nitrogen source against conventional N sources (yeast extract, peptone) was studied. The double increase was observed in propionic acid production while using yeast extract and peptone (0.29 ± 0.01 g/g), as against the use of only apple pomace extract (APE) (0.14 ± 0.01 g/g). Intensification of propionic acid fermentation was also achieved by increasing the pH control frequency of the culture medium from 24-(0.29 ± 0.01 g/g) to 12-hour intervals (30 °C) (0.30 ± 0.02 g/g) and by increasing the temperature of the culture from 30 to 37 °C (12-hour intervals of pH control) (0.32 ± 0.01 g/g). An important factor in improving the parameters of fermentation was the addition of biotin to the medium. The 0.2 mg/L dose of biotin allowed to attain 7.66 g/L propionate with a yield of 0.38 ± 0.03 g/g (12-hour intervals of pH control, 37 °C).

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%).

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.

Propionibacterium spp.-source of propionic acid, vitamin B12, and other metabolites important for the industry.

Bacteria from the Propionibacterium genus consists of two principal groups: cutaneous and classical. Cutaneous Propionibacterium are considered primary pathogens to humans, whereas classical Propionibacterium are widely used in the food and pharmaceutical industries. Bacteria from the Propionibacterium genus are capable of synthesizing numerous valuable compounds with a wide industrial usage. Biomass of the bacteria from the Propionibacterium genus constitutes sources of vitamins from the B group, including B12, trehalose, and numerous bacteriocins. These bacteria are also capable of synthesizing organic acids such as propionic acid and acetic acid. Because of GRAS status and their health-promoting characteristics, bacteria from the Propionibacterium genus and their metabolites (propionic acid, vitamin B12, and trehalose) are commonly used in the cosmetic, pharmaceutical, food, and other industries. They are also used as additives in fodders for livestock. In this review, we present the major species of Propionibacterium and their properties and provide an overview of their functions and applications. This review also presents current literature concerned with the possibilities of using Propionibacterium spp. to obtain valuable metabolites. It also presents the biosynthetic pathways as well as the impact of the genetic and environmental factors on the efficiency of their production.