1. Research based at the Prague University of Chemistry and Technology
Project Manager: Doc. Ing. Evgeny Razumov, DrSc
SPECTROSCOPUS CHARACTERISATION OF CHAGA (INONOTUS OBLIQUUS) PREPARATIONS: A CONTRIBUTION OF PHENOLICS AND POLYSACCHARIDES
Authors:
Lucie Třešnáková, Roman Bleha, Andrej Sinica, Jana Čopíková
Department of Carbohydrates and Cereals, Faculty of Food and Biochemical Technology, University of Chemistry and Technology Prague;
Evgeny Razumov
Faculty of Forestry and Wood Sciences, Czech University of Life Sciences, Prague, Czech Republic.
Participation in a scientific conference:
Proceedings of 14 International Conference on Polysaccharides-Glycoscience.
Prague
7-9 November 2018
Development of methods for identifying medicinal mushrooms using an infrared spectrometer.
Medicinal mushrooms such as reishi (Ganoderma lucidum), shiitake (Lentinula edodes), and chaga (Inonotus obliquus) are widely known for their therapeutic properties. Modern science is actively researching ways to identify them to ensure the quality and safety of biologically active substances. One of the promising methods for analyzing medicinal mushrooms is infrared spectroscopy (IR spectroscopy). This method allows for the rapid and accurate detection of chemical compounds such as polysaccharides, triterpenoids, and phenolic compounds that play a key role in the medicinal properties of mushrooms.
An infrared spectrometer works by using infrared radiation, which interacts with the molecules of a substance, causing them to vibrate. Different functional groups in the molecules absorb infrared radiation at different wavelengths, which allows for a unique absorption spectrum. The absorption spectra of fungi create a kind of “fingerprint” that can be used for their precise identification.
Infrared spectroscopy methods are used to determine the composition and identify active substances in fungi. For example, polysaccharides are characterized by absorptions in the range of 3000-3500 cm⁻¹, reflecting the presence of hydroxyl groups. These compounds are known to have immunomodulatory properties. Triterpenoids, which are antioxidants, are characterized by absorption bands in the range of 1500-1800 cm⁻¹, corresponding to carbonyl groups.
One of the key aspects of using an infrared spectrometer is the need to create a database of spectra of various fungi. This allows you to automate the process of identifying and comparing new samples with already known ones. Modern algorithms for analyzing spectral data, such as machine learning and artificial neural networks, can significantly improve the accuracy and speed of identification.
With the help of IR spectrometers, you can also track changes in the composition of fungi depending on their growth conditions. This opens up opportunities to optimize cultivation conditions to increase the concentration of certain biologically active substances. For example, different light and temperature conditions can affect the level of accumulation of triterpenoids in Ganoderma mushrooms.
In addition, infrared spectroscopy allows us to assess the effect of mushroom processing methods on the content of active components. This is especially important in the pharmaceutical industry, where thermal and chemical processing of raw materials can significantly change their composition.
IR spectrometers can also be used to detect counterfeiting. Often, unscrupulous manufacturers add artificial substances to drugs that imitate natural mushroom components. Spectral data will help to identify such counterfeits at the early stages of analysis.
One example of the successful application of IR spectroscopy is the study of the composition of mushrooms in traditional Chinese medicine. In mushrooms such as chaga, reishi and shiitake, unique combinations of active compounds have been identified that play a key role in their medicinal properties. These data make it possible to standardize and regulate the production of mushroom-based drugs. In conclusion, the use of infrared spectroscopy for the identification and analysis of medicinal mushrooms is a promising direction in mycological science. This method provides accurate, fast and non-destructive analysis of biologically active substances, which is especially important in medicine and the pharmaceutical industry.
2. International scientific project. The aim of the project is to identify active substances in medicinal mushrooms.
Participation in a scientific conference:
IOP Conf. Series: Materials Science and Engineering (2019)
Project Manager: Doc. Ing. Evgeny Razumov, DrSc
INFLUENCE OF DEVELOPMENT CONDITIONS ON QUALITATIVE INDICATORS OF FUNGUS CHAGA
University of Life Sciences, Prague, Czech Republic.
Kazan National Research Technological University, Kazan.
Technical University in Zvolen. Slovak Republic.
3. Joint research with the Czech University of Life Sciences (Czech Republic); Technical University in Zvolen
Project Manager: Doc. Ing. Evgeny Razumov, DrSc
Wood Research 65(5): (2020)
833-842
EXTRACTION OF BETULIN FROM THE BIRCH BARK BALANCE AT PULP AND PAPER PRODUCTION
Health benefits of betulin: research and application in pharmaceuticals Introduction
Betulin is a natural compound extracted from birch bark, known for its diverse health benefits. The chemical structure of betulin makes it unique among natural substances. In recent years, it has been actively studied in the field of pharmaceuticals due to its potential therapeutic effects, such as anti-inflammatory, antiviral and antitumor effects.
Composition of betulin
Betulin is a triterpenoid containing a pentacyclic structure that includes five carbon cycles and hydroxyl groups. Its molecule is based on lupane, which gives betulin high stability and biochemical activity. Betulin also contains various functional groups that can change its chemical and biological properties, making it suitable for modification to create new drugs.
Healing properties of betulin
1. Anti-inflammatory effect. Studies have shown that betulin can reduce the activity of inflammatory processes in the body. It inhibits key inflammatory mediators such as cyclooxygenase and interleukins, making it promising for the treatment of inflammatory diseases such as arthritis and dermatitis.
2. Antiviral action. Betulin exhibits antiviral activity against a number of viruses, including HIV, herpes, and influenza. It prevents viruses from penetrating cells and replicating, making betulin a potential tool for developing antiviral drugs.
3. Antibacterial action. Betulin is able to suppress the growth of some pathogenic bacteria, including antibiotic-resistant strains. This property makes it promising for use in the fight against infections, especially in the context of the growing problem of antibiotic resistance.
4. Antitumor action. One of the most studied properties of betulin is its ability to suppress the growth of cancer cells. Studies have shown that betulin induces apoptosis (programmed cell death) in various types of tumors, including skin, liver, lung and breast cancer. This is due to its effect on mitochondrial pathways and inhibition of cell proliferation.
5. Hepatoprotective action. Betulin has the ability to protect liver cells from damage caused by toxins, alcohol and drugs. Its use can be useful in the treatment of liver diseases such as hepatitis and cirrhosis.
6. Antioxidant action. Betulin acts as a powerful antioxidant, protecting cells from oxidative stress caused by free radicals. This property is important for preventing aging, the development of cardiovascular diseases and neurodegenerative disorders.
Use of betulin in pharmaceuticals
1. Preparations for the treatment of skin diseases. In dermatology, betulin is widely used to treat inflammatory and infectious skin diseases such as eczema, psoriasis, and acne. Betulin-based creams and ointments help reduce inflammation, improve tissue regeneration, and accelerate wound healing.
2. Anticancer drugs. Promising drugs for the treatment of oncological diseases are created based on betulin. Its ability to induce apoptosis in cancer cells and have minimal impact on healthy cells makes betulin an interesting object for the development of targeted antitumor agents.
3. Hepatoprotectors. Betulin-based drugs can be used to prevent and treat liver diseases such as hepatitis and fatty liver disease. They promote the restoration of hepatocytes and prevent their damage under the influence of toxic substances.
4. Antiviral drugs. Betulin, having a wide range of antiviral activity, can be used in drugs for the prevention and treatment of viral infections, including HIV and influenza. Research is ongoing to develop effective new-generation antiviral drugs based on it.
5. Cardiovascular disease prevention products. The antioxidant and anti-inflammatory properties of betulin make it promising for developing products aimed at reducing the risk of cardiovascular diseases. Products based on it can help prevent atherosclerosis, coronary heart disease, and other pathologies.
6. Immunostimulating drugs. Betulin can be used as part of immunomodulatory drugs, since its antiviral and anti-inflammatory properties help enhance the body’s immune response. This is especially important in conditions of increased epidemiological stress.
Conclusion
Betulin, due to its multifaceted biological properties, has significant potential for use in pharmaceuticals. Studies show its effectiveness in combating inflammatory processes, viral infections, tumors, and liver diseases. Further study of betulin and its derivatives can lead to the creation of new drugs for the treatment of a wide range of diseases.
4. The research was conducted at the Czech University of Life Sciences in Prague.
Project Manager: Doc. Ing. Evgeny Razumov, DrSc
Participation in a scientific conference:
IOP Conf. Series: Materials Science and Engineering (2020)
Cryogenic Milling Technology for Medicinal Mushrooms: Innovations to Improve Absorption and Efficacy
Medicinal mushrooms such as reishi, shiitake and chaga are important sources of bioactive compounds used in traditional and modern medicine. The main bioactive components of mushrooms, including polysaccharides, proteins and vitamins, exhibit a wide range of health benefits, from strengthening the immune system to anti-cancer properties. However, the effectiveness of these compounds largely depends on their bioavailability, i.e. the body’s ability to absorb them after consumption. Traditionally, the milling of mushrooms into powder often limits this bioavailability, as many biomolecules remain in a stable form and are difficult for digestive enzymes to access. In this context, cryogenic milling technology has emerged as a key innovation to improve the absorption and therapeutic efficacy of mushroom powders.
Cryogenic milling is a process in which raw materials are crushed at ultra-low temperatures, often in the range of -50 to -196°C. By using liquid nitrogen or other cryogenic liquids, the material is frozen, making it more fragile. This allows for finer and more efficient grinding, down to the level of micron particles. This method eliminates overheating, which can destroy important biomolecules, and preserves all the active components of the mushrooms in their original state. One of the main advantages of cryogenic grinding is the increased bioavailability of mushroom powders. This is due to a change in the physical structure of the raw material: cryogenic processing destroys the cell walls, which consist of chitin, a polymer that is the main obstacle to the release of active substances. Chitin is poorly absorbed by the human body, and its destruction allows the release of proteins, polysaccharides and other components. As a result, the surface area of the particles increases, which accelerates and facilitates their interaction with enzymes in the gastrointestinal tract.
In addition, an important feature of cryogenically ground powders is the change in protein structures. Under the influence of low temperatures, mushroom proteins undergo denaturation, that is, their complex spatial structures disintegrate, and they are transformed into simpler molecules – amino acids. This is important, since the human body absorbs amino acids more easily than proteins in their original form. Amino acids can quickly enter into metabolic processes, which enhances the pharmacological effect of the mushroom, whether it is stimulation of the immune system or regulation of inflammatory processes.
Comparison of traditional mushroom powder with cryogenically ground powder reveals significant differences in their digestibility and effectiveness. Conventional powder has large particles with intact cell walls, which makes it difficult to access active substances. Many polysaccharides and proteins remain in an indigestible form, and the chitinous shell prevents the release of biomolecules. As a result, the therapeutic effect of such a powder can be limited, and a significantly larger dose is required to achieve the desired effect. In the case of cryogenically ground powder, these problems are minimized. By reducing the particle size to micron values and destroying chitinous cell walls, cryogenic powder has significantly greater bioavailability. The conversion of proteins into amino acids also accelerates their absorption and involvement in metabolic processes. This makes cryogenic powder a more effective remedy, allowing for a lower dosage and achieving a more pronounced therapeutic effect.
In addition to improved bioavailability, cryogenic powder preserves the activity of heat-sensitive compounds such as enzymes, vitamins and antioxidants. During traditional grinding, which involves heating, these components can be partially destroyed, which reduces the overall effectiveness of the product. Cryogenic technology, on the contrary, ensures maximum preservation of all useful substances, which is especially important for products used in pharmaceuticals and nutraceuticals.
Practical studies of cryogenically ground mushrooms show higher pharmaceutical activity compared to conventional powders. In clinical trials, patients taking cryogenic powder demonstrated faster and more sustainable results in the treatment of various diseases, such as immune disorders, inflammation and cancer. This indicates that cryogenic technology opens up new horizons for the use of medicinal mushrooms in medicine. Thus, cryogenic grinding of mushrooms is a promising technology that significantly increases the effectiveness of mushroom-based drugs. Due to the destruction of chitin, the conversion of proteins into amino acids and the preservation of heat-sensitive substances, cryogenically ground powders have clear advantages over traditional ones. In the future, we can expect further dissemination of this technology in the pharmaceutical and food industries, which will maximize the potential of medicinal mushrooms for human health.
5. Research into Growing Medicinal Mushrooms Lion's Mane, Shiitake and Reishi in Climate Chambers (2024)
Project Manager: Martina Fiedlerová
Medicinal mushrooms such as Lion’s Mane (Hericium erinaceus), Shiitake (Lentinula edodes) and Reishi (Ganoderma lucidum) are known for their beneficial properties, including immunomodulatory and anti-inflammatory effects. Growing these mushrooms in controlled conditions such as climate chambers allows for an optimized cultivation process and an increase in the content of bioactive compounds. Research in this area shows that maintaining precise microclimate parameters such as temperature, humidity, light and carbon dioxide levels are key to success.
For Lion’s Mane, which grows on wood in temperate forests, temperatures in the 20 to 25 °C range and high humidity (80–90%) are important. The mushroom requires a certain light intensity at the fruiting stage, which is close to the natural conditions of its habitat in forests. Studies have shown that creating such conditions promotes the accumulation of hericenones and erinarins – compounds that promote the regeneration of nerve cells.
Shiitake, traditionally grown in East Asia, prefers temperatures within the range of 18-22 °C at the mycelial growth stage and about 15-18 °C at the fruiting stage. Humidity should be at the level of 85-90%. Research also emphasizes the importance of the light cycle: mushrooms grow better with alternating light and dark periods, which imitates natural day and night. This promotes maximum synthesis of lentinan – a polysaccharide with a pronounced antitumor effect.
Reishi, unlike the two previous species, grows in warmer and more humid climates. Temperatures ranging from 25 to 30 °C, humidity around 85–90% and low light levels allow this mushroom to produce the maximum amount of triterpenoids and polysaccharides with immunomodulatory and antioxidant properties. Particular attention is paid to controlling the CO2 concentration: increased levels of carbon dioxide in the chamber stimulate the growth of fruiting bodies, increasing the yield of bioactive substances.
One of the key challenges in cultivating medicinal mushrooms is creating optimal conditions for each stage of their life cycle. For example, the successful growth of Lion’s Mane mycelium requires a lower temperature (around 20 °C), while for the formation of fruiting bodies it must be increased to 24–25 °C. Humidity and oxygen levels are also regulated at different stages: high humidity and limited oxygen access are important during the incubation phase, while increased oxygen levels should be used during the fruiting stage. Research shows that synchronizing climate parameters with the natural life cycle of mushrooms plays an important role in the accumulation of valuable substances. For example, for Lion’s Mane, creating moderately warm and humid conditions that imitate the rainy season leads to an increase in the synthesis of erinarins, which are useful for the treatment of neurodegenerative diseases. While shiitake grows better with changing temperature regimes that imitate spring and autumn periods.
It has been established that regulation of illumination affects not only the growth of fruiting bodies, but also the accumulation of active substances. For example, reishi produces triterpenoids better under minimal illumination, since this mushroom naturally grows in shady places. At the same time, shiitake requires a moderate level of light to stimulate the production of lentinan.
An important part of the research is the development of automated systems for monitoring and regulating the microclimate in climatic chambers. Modern technologies make it possible to control changes in temperature, humidity, illumination and CO2 concentration in real time, which increases the stability of growing conditions. The use of such systems allows minimizing the human factor and significantly increases the productivity of mushrooms.
The use of climatic chambers also opens up the possibility of conducting experiments on changing the composition of substrates, which can significantly affect the biochemical composition of mushrooms. For example, adding enriched organic components to the substrate for shiitake helps to increase the content of lentinan and other useful polysaccharides. In the case of Lion’s Mane, studies show that wood substrates from certain tree species stimulate the formation of hericenones. Ultimately, research into growing medicinal mushrooms in climate chambers is aimed at achieving maximum biological activity of the mushrooms by fine-tuning conditions that mimic natural cycles. By creating an optimal microclimate, the content of medicinal substances can be significantly increased, making mushrooms a more valuable source for medicine and pharmaceuticals.
6. Joint research with Technical University in Zvolen
Project Manager: Doc. Ing. Evgeny Razumov, DrSc
Effect of habitat on quantitative indices of muscimol in red fly agaric (amanita muscaria)
(2024)
The red fly agaric (Amanita muscaria) is a mushroom widely known for its psychoactive properties due to the presence of such substances as muscimol and ibotenic acid. Muscimol is the main component causing the psychoactive effect, and its concentration can vary significantly depending on various environmental factors. Studies devoted to the effect of habitat on the level of muscimol in the red fly agaric play an important role in better understanding the mechanisms of its accumulation.
The first factor that was studied in the context of the effect on muscimol content is the type of soil. Fungi, as mycorrhizal organisms, form symbiotic relationships with trees, and the composition of the soil can affect the growth of the fungus and the synthesis of its secondary metabolites. Studies have shown that red fly agarics growing in poor sandy soils may contain less muscimol compared to mushrooms growing in more fertile soils rich in organic matter.
Another important environmental factor is the humidity of the environment. Fly agarics growing in high humidity conditions tend to have increased biomass, which in turn may contribute to higher levels of muscimol accumulation. The content of the active substance can also be affected by the temperature of the environment. In moderate temperatures, optimal synthesis of muscimol is observed, while extremely high or low temperatures can inhibit this process.
Illumination and access to sunlight also affect the concentration of muscimol. In places with moderate shade, fly agarics showed higher concentrations of the active substance than in strong sunlight, which is probably due to the protective mechanisms of the fungus against ultraviolet radiation.
The age of the mushroom is another parameter that should be taken into account when studying the quantitative indicators of muscimol. It was found that more mature fly agarics have significantly higher levels of muscimol than young mushrooms. This may be due to the cumulative effect during the growth and development of the fungus.
Researchers also pay attention to the soil microbiota, which can affect the chemical composition of the fungi. The interaction between fungi and soil microorganisms, such as bacteria and other fungi, can both stimulate and suppress the production of muscimol. Experiments have shown that some types of bacteria present in the soil can promote increased synthesis of this substance.
Geographical location and climatic conditions play an important role in the muscimol content of fly agarics. Mushrooms collected in regions with harsher climatic conditions, such as in northern latitudes, often contain more muscimol than mushrooms growing in mild climatic zones. This may be due to the adaptation of the fungus to extreme environmental conditions. The study of the influence of soil and environmental pollution on the muscimol content in mushrooms also seems to be a promising direction. In contaminated soil conditions, the level of metabolites may decrease, which indicates the potential sensitivity of the red fly agaric to the presence of toxic substances in the environment. In conclusion, the habitat has a significant impact on the quantitative indicators of muscimol in the red fly agaric. Soil type, humidity, temperature, illumination, age of the mushroom and soil microbiota are all factors that affect the level of this psychoactive substance. Ecological studies in this area can help to better understand the conditions that promote the synthesis of muscimol, which is important both for scientific research and for the potential use of the red fly agaric for medicinal and pharmacological purposes.
In Canada, several years ago, the red fly agaric (amanita muscaria) was officially recommended for the production of dietary supplements. https://webprod.hc-sc.gc.ca/nhpid-bdipsn/searchIngred
7. Growing Medicinal Mushrooms on Tree Stumps: Ecological and Economic Benefits
Project Manager: Martina Fiedlerová
Growing functional mushrooms on tree stumps is a promising method of using forest resources that benefits both the ecosystem and humans. This approach is based on the fact that tree stump mushrooms are saprophytes, meaning they feed on dead organic matter, promoting wood decomposition and accelerating natural forest restoration processes. Some of the most popular types of mushrooms for stump cultivation are reishi (Ganoderma lucidum) and turkey tail (Trametes versicolor). These species not only provide ecological benefits, but also have important medicinal properties, making their cultivation attractive from a commercial point of view.
The Role of Tree Stump Mushrooms in the Ecosystem
Fungi growing on tree stumps play a key role in forest ecosystems, helping to decompose dead wood and convert organic material into nutrients that are then used by other organisms. After forests are cleared, tree stumps are often left in the soil, where they are uprooted and composted. However, instead of removing these stumps, you can use them as a substrate for cultivating mushrooms.
Benefits of cultivating mushrooms on stumps:
- Forest restoration. Fungi promote the decomposition of dead wood, accelerating soil restoration and preventing erosion. This improves the conditions for new trees to grow.
- Filling ecological niches. Fungi can fill empty niches, preventing the spread of harmful organisms such as insect pests and pathogenic fungi.
- Sustainable use of resources. Stumps that would otherwise be considered waste can be used to produce valuable products.
- Pest control. Some fungi species, such as Trametes versicolor, can compete with wood pathogens, reducing their numbers.
Economic and medicinal value of mushrooms
Growing mushrooms on tree stumps can bring significant economic benefits, especially if we are talking about species with medicinal properties. Two examples of such mushrooms are reishi and turkey tail.
Reishi (Ganoderma lucidum)
Reishi mushroom, known in traditional Chinese medicine as the “mushroom of immortality”, is famous for its immunomodulatory, antioxidant and antitumor properties. Growing it on tree stumps is not only an efficient use of resources, but also a way to produce raw materials for the pharmaceutical industry.
The method of growing reishi on tree stumps involves inoculating the mycelium of the fungus into wood, preferably hardwoods such as oak or maple. Tree stumps from deforestation are an ideal environment for the growth of this type of mushroom. The mycelium develops inside the wood, decomposing it and forming fruiting bodies that can be collected and processed.
Turkey Tail (Trametes versicolor)
Trametes versicolor, known as “turkey tail” due to the characteristic shape and color of the fruiting bodies, also has significant medicinal properties. This mushroom is widely used in Eastern medicine as an immunomodulator and antitumor agent. Turkey tail extracts contain polysaccharide-K (PSK), which is used to support the immune system in cancer patients.
As in the case of Reishi, tree stumps after deforestation can be effectively used for the cultivation of Trametes versicolor. This mushroom is especially valuable, since its fruiting bodies develop quickly on old wood, promoting its decomposition and providing additional income for forestry enterprises.
Technology of inoculation of mushrooms on stumps
The process of growing mushrooms on stumps begins with the preparation of the mushroom mycelium. This mycelium is then inoculated into the stump, which has already undergone partial decomposition or remains fresh after the tree has been cut down. Inoculation can be done in several ways, such as using wooden dowels soaked in mycelium or applying mycelial mass to the surface of the stump.
- Preparing the stump. The stumps should preferably be hardwood and not too rotten. Fresh stumps that have been cut within a year are best.
- Inoculation. The mycelium is introduced into the stump through holes made, which are filled with mycelial dowels or granules. Mycelial blocks can also be used, which are attached to the surface of the stump.
- Controlling the conditions. After inoculation, it is important to maintain humidity and optimal conditions for mycelial growth. If necessary, stumps can be protected from direct sunlight and high ambient humidity can be maintained.
- Harvesting. Depending on the mushroom species and growing conditions, fruiting bodies can begin to form 6–18 months after inoculation. The harvest is hand-picked, and the mycelium continues to process the wood, ensuring long-term productivity.
Environmental benefits
In addition to the economic benefits, growing mushrooms on stumps makes a significant contribution to sustainable forestry. Mushrooms help decompose organic material, which speeds up the natural regeneration of forests, improving soil quality and creating optimal conditions for the growth of new trees. This also reduces the need for mechanical processing and uprooting of stumps, which can be a costly and labor-intensive process.
Moreover, mushrooms can prevent the spread of pests and diseases. By filling empty ecological niches, mushrooms compete with pathogens and insects, thereby protecting the remains of forest stands.
Conclusion
Growing mushrooms on stumps is an example of a symbiotic approach to the use of natural resources that benefits both the ecosystem and humans. Medicinal mushrooms such as reishi and turkey tail not only contribute to wood decomposition and forest restoration, but also provide valuable raw materials for the pharmaceutical industry. This makes mushroom cultivation on tree stumps a promising direction for forestry aimed at sustainable development and ecosystem restoration.
