Characteristics of exopolysaccharides from Paecilomyces hepiali and their simulated digestion and fermentation in vitro by human intestinal microbiota.

The metabolic process of polysaccharides in gastrointestinal digestions and the effects of the resulting carbohydrates on the composition of gut microbes are important to explore their prebiotic properties. Therefore, the purpose of this study was to investigate the simulated digestion and fecal fermentation in vitro of three fractions (PHEPSs-1, PHEPSs-2 and PHEPSs-3) purified from the crude exopolysaccharides of Paecilomyces hepiali HN1 (PHEPSs) and to explore the potential prebiotic mechanisms. The three purified fractions were characterized by HPLC, UV, FT-IR, SEM and AFM, and they were all of galactoglucomannan family with molecular weight of 178, 232 and 119 kDa, respectively. They could resist the simulated gastrointestinal digestions, but they were metabolized in fecal fermentation in vitro. Furthermore, the mannose in PHEPSs showed a higher utilization rate than that of glucose or galactose. The proliferation effects of PHEPSs on Bifidobacterium and Lactobacillus were weaker significantly than those of fructooligosaccharides before 12 h of fecal fermentation, but stronger after 24 h of fecal fermentation. Meanwhile, higher levels of short-chain fatty acids were found in PHEPSs groups when the fecal fermentation extended to 36 h. Therefore, PHEPSs are expected to have a potent gut healthy activity and can be explored as functional food ingredients.

Structural diversity and biological activities of indole-diterpenoids from Penicillium janthinellum by co-culture with Paecilomyces formosus.

Co-culturing the marine-derived fungi Penicillium janthinellium with Paecilomyces formosus led to the isolation of nine new indole-diterpenes, janthinellumines A-I (1-9), along with twelve known analogues (10-21). The chemical structures including their absolute configurations of them were assigned by the analysis of extensive spectroscopic data and calculated ECD and VCD methods. These indole-diterpenoids displayed extensive biological activities, including anti-influenza A virus, protein tyrosine phosphatase (PTP) inhibitory, and anti-Vibrio activities. Among them, the anti-influenza mechanism of compounds 1, 2, and 7 was further investigated using neuraminidase inhibitory assay, molecular docking, and reverse genetics methods, suggesting that 1, 2, and 7 could interact with Arg371 of the viral neuraminidase. The structure-activity relationship (SAR) of PTPs inhibitory activity for indole-diterpene derivatives (1, 2, 4, 5, 9-16, and 19-21) was also summarized.

Locating and Quantifying Carbon Steel Corrosion Rates Linked to Fungal B20 Biodiesel Degradation.

Fungi that degrade B20 biodiesel in storage tanks have also been linked to microbiologically influenced corrosion (MIC). A member of the filamentous fungal genus Paecilomyces and a yeast from the genus Wickerhamomyces were isolated from heavily contaminated B20 storage tanks from multiple Air Force bases. Although these taxa were linked to microbiologically influenced corrosion in situ, precise measurement of their corrosion rates and pitting severity on carbon steel was not available. In the experiments described here, we directly link fungal growth on B20 biodiesel to higher corrosion rates and pitting corrosion of carbon steel under controlled conditions. When these fungi were growing solely on B20 biodiesel for carbon and energy, consumption of FAME and n-alkanes was observed. The corrosion rates for both fungi were highest at the interface between the B20 biodiesel and the aqueous medium, where they acidified the medium and produced deeper pits than abiotic controls. Paecilomyces produced the most corrosion of carbon steel and produced the greatest pitting damage. This study characterizes and quantifies the corrosion of carbon steel by fungi that are common in fouled B20 biodiesel through their metabolism of the fuel, providing valuable insight for assessing MIC associated with storing and dispensing B20 biodiesel. IMPORTANCE Biodiesel is widely used across the United States and worldwide, blended with ultra-low-sulfur diesel in various concentrations. In this study, we were able to demonstrate that the filamentous fungus Paecilomyces AF001 and the yeast Wickerhamomyces SE3 were able to degrade fatty acid methyl esters and alkanes in biodiesel, causing increases in acidity. Both fungi also accelerated the corrosion of carbon steel, especially at the interface of the fuel and water, where their biofilms were located. This research provides controlled, quantified measurements and the localization of microbiologically influenced corrosion caused by common fungal contaminants in biodiesel fuels.

Improvement of cordycepin production by an isolated Paecilomyces hepiali mutant from combinatorial mutation breeding and medium screening.

Cordycepin is a major bioactive compound found in Cordyceps sinensis that exhibits a broad spectrum of biological activities. Here a Paecilomyces hepiali OR-1 strain was initially isolated from plateau soil for the bioproduction of cordycepin. Subsequently, strain modification including 60Co γ-ray and ultraviolet irradiation were employed to increase the cordycepin titer, resulted in a high-yield mutant strain P. hepiali ZJB18001 with the cordycepin content of 0.61 mg/gDCW, showing a 2.3-fold to that from the wild strain (0.26 mg/gDCW). Furthermore, medium screening based on Box-Behnken design and the response surface methodology facilitated the enhancement of cordycepin yield to the value of 0.96 mg/gDCW at 25 °C for 5 days in submerged cultivation with an optimized medium composition. The high cordycepin yield, rapid growth rate and stable genetic characteristics of P. hepiali ZJB18001 are beneficial in terms of costs and time for the industrialization of cordycepin production.

Exocellular polysaccharides extracted from mangrove fungus Paecilomyces Lilacinuson present anti-HSV-1 activity in mice.

This study examined the anti-HSV-1 activity of EPS extracts isolated from mangrove fungus Paecilomyces Lilacinuson after intraperitoneal administration in mice. Mice were experimentally infected with HSV-1 intracranially and treated intraperitoneally with three different doses of EPS extract (6 g/Kg, 8 g/Kg, and 10 g/Kg) for 7 days. One group of 15 mice was infected with HSV-1 but did not receive any treatment, while another group of 15 mice was mock-infected to remain a control group. Animals were observed twice a day for 14 days after virus infection, searching for clinical signs of weight loss, piloerection, isolation, or retardation movement. Compared with the mock-infected group, mortality was significantly increased (p < 0.05) in the virus-infected group and the groups that received 6 g/Kg and 8 g/Kg EPS extract. Interestingly, no significant differences in mortality were found between the 10 g/Kg EPS extract and the mock-infected group. Mortality in the 10 g/Kg EPS extract group was substantially improved compared with virus-infected(p < 0.05). Additionally, EPS extracts inhibited HSV-1 replication in the mice brain in a dose-dependent manner. Furthermore, the extracts decreased NF-κB protein and mRNA expression and the production of TNF-α in HSV-1-infected mice brain tissue. These effects were also dose-dependent. Our findings suggest that the EPS extract may be a potential candidate for developing an antiviral drug against HSV-1.

Identification of the pigment and its role in UV resistance in Paecilomyces variotii, a Chernobyl isolate, using genetic manipulation strategies.

Fungi produce secondary metabolites that are not directly involved in their growth, but often contribute to their adaptation to extreme environmental stimuli and enable their survival. Conidial pigment or melanin is one of the secondary metabolites produced naturally by a polyketide synthesis (PKS) gene cluster in several filamentous fungi and is known to protect these fungi from extreme radiation conditions. Several pigmented or melanized fungi have been shown to grow under extreme radiation conditions at the Chernobyl nuclear accident site. Some of these fungi, including Paecilomyces variotii, were observed to grow towards the source of radiation. Therefore, in this study, we wanted to identify if the pigment produced by P. variotii, contributes to providing protection against radiation condition. We first identified the PKS gene responsible for synthesis of pigment in P. variotii and confirmed its role in providing protection against UV irradiation through CRISPR-Cas9 mediated gene deletion. This is the first report that describes the use of CRISPR methodology to create gene deletions in P. variotii. Further, we showed that the pigment produced by this fungus, was not inhibited by DHN-melanin pathway inhibitors, indicating that the fungus does not produce melanin. We then identified the pigment synthesized by the PKS gene of P. variotii, as a naptho-pyrone Ywa1, by heterologously expressing the gene in Aspergillus nidulans. The results obtained will further aid in understanding the mechanistic basis of radiation resistance.

Scorpionicidal activity of secondary metabolites from Paecilomyces sp. CMAA1686 against Tityus serrulatus.

INTRODUCTION: Urban pests pose enormous risks to human health. Control initiatives are carried out in regions of high infestation and incidence of accidents caused by scorpions OBJECTIVE: In this study, we aimed to analyze the anti-scorpionic activity of fungal isolates obtained from a cemetery in Brazil. MATERIALS AND METHODS: A total of thirteen fungi were subjected to a bioassay test against Tityus serrulatus, and the two isolates with the highest scorpionicidal activity were selected for molecular identification through sequencing of the ITS DNA hypervariable region and large-scale cultivation on liquid medium for secondary metabolite extraction. The crude extracts were partitioned by solid-phase extraction, and the resulting purified extracts were tested for anti-scorpionic activity. The extracts from one of the isolates presented better results and were submitted to UPLC-MS/MS. The metabolomics data were submitted to GNPS website for Molecular Networking and MASST searches. We also performed a MolNetEnhancer analysis to identify the chemical classes of the molecules found in the samples. RESULTS: The most promising fungal isolate was identified as Paecilomyces sp. CMAA1686 which has 98% of similarity to Paecilomyces formosus. The sub-fractions C and D had the best activity against the scorpions (54 and 32% mortality, respectively). Molecular Networking and MolNetEnhancer revealed a range of molecular classes in our extracts that are known to include bioactive metabolites from Paecilomyces species. CONCLUSIONS: The scorpionicidal activity of Paecilomyces sp. CMAA1686 and its secondary metabolites may provide new alternative compounds for biological and chemical control of scorpions from the species T. serrulatus. Paecilomyces sp. CMAA1686 is an isolate that has great potential for isolation of secondary metabolites.

Secondary Metabolites and Their Bioactivities Produced by Paecilomyces.

Paecilomyces, a common saprobic filamentous fungus, not only plays an important role in biological control, but also has applications in medicine, food, and environmental protection. In this paper, 223 secondary metabolites and their bioactivities from 13 known species and various unidentified strains of Paecilomyces are reviewed. Their structures can be described as polyketide, terpenoid, peptide, alkaloid, quinone, pyrone, sterol, and fatty acid. They have been demonstrated varying biological activities, including antimicrobial, antitumor, insecticidal, antiplasmodial, antimalarial, nematicidal, herbicidal, and enzyme-inhibiting. This review provides a comprehensive overview of secondary metabolites and their biological activities from strains of Paecilomyces.

Cadinane- and drimane-type sesquiterpenoids produced by Paecilomyces sp. TE-540, an endophyte from Nicotiana tabacum L., are acetylcholinesterase inhibitors.

Sesquiterpenoids with diverse skeleton types are regarded as potential lead compounds in pharmacological and other applications. Herein, we report the discovery of two new cadinane-type sesquiterpenoids, paecilacadinol A (1) and B (2); two new drimane-type sesquiterpenoids, ustusol D (3) and ustusol E (4); and six known analogs (5-10) from the endophytic fungus Paecilomyces sp. TE-540, enriching the structural diversity of naturally occurring sesquiterpenoids. Their planar structures were determined on the basis of detailed interpretation of 1D and 2D NMR spectroscopy and HRESIMS data, while their stereochemical structures were established by X-ray crystallographic analyses for 1 and 3-8 and theoretical calculations for 2. Notably, compounds 1 and 2 represent novel examples of cadinane-type sesquiterpenoids with ether bonds formed by intramolecular dehydration. Compounds 5 and 6 showed moderate activities against acetylcholinesterase (AChE), with IC50 values of 43.02 ± 6.01 and 35.97 ± 2.12 µM, respectively. Docking analysis predicted that 5 bound well in the catalytic pocket of AChE via hydrophobic interactions with Trp84, Gly117, Ser122, and Tyr121 residues, while 6 was located with Asp72 and Ser122 residues.

Induced terreins production from marine red algal-derived endophytic fungus Aspergillus terreus EN-539 co-cultured with symbiotic fungus Paecilomyces lilacinus EN-531.

The coculture of marine red algal-derived endophytic fungi Aspergillus terreus EN-539 and Paecilomyces lilacinus EN-531 induced the production of a new terrein derivative, namely asperterrein (1) and a known dihydroterrein (2), which were not detected in the axenic cultures of both strains. The production of the known secondary metabolites terrein (3), butyrolactone I (4), and dankasterone (6), derived from A. terreus EN-539, were depressed significantly in the coculture. Compounds 1-3 exhibited inhibitory activity against Alternaria brassicae, Escherichia coli, Physalospora piricola, and Staphylococcus aureus with MIC values ranging from 4 to 64 µg ml-1.

Solid-state Co-cultivation of Bacillus subtilis, Bacillus mucilaginosus, and Paecilomyces lilacinus Using Tobacco Waste Residue.

Agro-industrial wastes are excellent sources for solid-state culture to produce spores of microorganisms, whereas microbial co-cultivation is not fully exploited in solid-state culture. In this work, the co-cultivation of different strains of Bacillus subtilis, and three microbes of B. subtilis, Bacillus mucilaginosus, and Paecilomyces lilacinus was studied using a solid medium only composed of water and tobacco waste residue after extraction of nicotine and solanesol. The influences of matrix thickness, moister, temperature, and ratio of three microbes in seed on the cell growth and spore formation were studied. The maximum viable cells and spores of each microbe reached 1013 cfu/g when cultured alone at 30 °C in a medium containing 58.3% moisture. Co-cultivation of microbes stimulated cell growth and maximum viable cells of each microbe reached 1014 cfu/g, while spore production was inhibited and decreased to 1011 cfu/g. With decreasing amount of P. lilacinus in seed, total amount of spores was increased. When the seed with a ratio of 6:3:1 for B. mucilaginosus, B. subtilis, and P. lilacinus was inoculated, the total amount of spores reached 4.14 × 1012 cfu/g and the ratio was 1.7:0.7:1. These results indicate the potential of solid-state cultivation in the high production of spores from tobacco waste residue at low cost.

Chitosan production from Paecilomyces saturatus using three monosaccharides via mixture design.

In this study, we demonstrate that chitosan is produced from Paecilomyces saturatus fungi using ternary monosaccharide carbon sources liquid cultivation via mixture design strategy. Sixteen experiments were carried out to obtain regression equations of fungal dry mycelial biomass (W), chitosan ratio (R), and deacetylation degree (DD) for plotting contour lines. Contour lines reveal that the maximum W, R, and DD can be simultaneously obtained in cultivated media containing 20% glucose, 60% fructose and 20% mannitol rather than pure monosaccharide cultivation. Three additional confirmation experiments based on the maximum FuCS deacetylation degree had been performed to confirm to be 92.3% via Fourier-transform infrared spectra. Accordingly, FuCS possessed much better anti-microbial activity on E. coli than commercial chitosan (CrCS). Meanwhile, X-ray diffraction results confirmed that FuCS possessed both α and γ crystalline peaks while CrCS possessed only α crystalline peak, being collaborated with thermogravimetric analysis results. The superior FuCS was obtained by using ternary monosaccharides system in fungal culture via mixture design for the first time. This study provides a new approach to produce chitosan from fungal cultivation by using the mixture design strategy.

Chemical Constituent Profiling of Paecilomyces cicadae Liquid Fermentation for Astragli Radix.

Astragli Radix (AR) is one of the most popular traditional Chinese medicines with chemical constituents including flavonoids and saponins. As recently evidenced, some fungi or their fermentation liquid may have the potential to affect the bioactive constituents and different pharmacological effects of AR. Thus, the composition of fermented AR (FAR) produced by Paecilomyces cicadae (Miquel) Samson in liquid-state fermentation was investigated using a UHPLC-LTQ-Orbitrap mass spectrometer in both positive and negative ion modes. Firstly, the MSn data sets were obtained based on a data-dependent acquisition method and a full scan-parent ions list-dynamic exclusion (FS-PIL-DE) strategy. Then, diagnostic product ions (DPIs) and neutral loss fragments (NLFs) were proposed for better constituent detection and structural characterization. Consequently, 107 constituents in total, particularly microconstituents in FAR and AR, were characterized and compared in parallel on the same LTQ-Orbitrap instrument. Our results indicated that AR fermentation with Paecilomyces significantly influenced the production of saponins and flavonoids, especially increasing the content of astragaloside IV. In conclusion, this research was not only the first to show changes in the chemical components of unfermented AR and FAR, but it also provides a foundation for further studies on the chemical interaction between microbiota and AR.

Fungal Dirigent Protein Controls the Stereoselectivity of Multicopper Oxidase-Catalyzed Phenol Coupling in Viriditoxin Biosynthesis.

Paecilomyces variotii produces the antibacterial and cytotoxic ( M)-viriditoxin (1) together with a trace amount of its atropisomer ( P)-viriditoxin 1'. Elucidation of the biosynthesis by heterologous pathway reconstruction in Aspergillus nidulans identified the multicopper oxidase (MCO) VdtB responsible for the regioselective 6,6'-coupling of semiviriditoxin (10), which yielded 1 and 1' at a ratio of 1:2. We further uncovered that VdtD, an α/ß hydrolase-like protein lacking the catalytic serine, directs the axial chirality of the products. Using recombinant VdtB and VdtD as cell-free extracts from A. nidulans, we demonstrated that VdtD acts like a dirigent protein to control the stereoselectivity of the coupling catalyzed by VdtB to yield 1 and 1' at a ratio of 20:1. Furthermore, we uncovered a unique Baeyer-Villiger monooxygenase (BVMO) VdtE that could transform the alkyl methylketone side chain to methyl ester against the migratory aptitude.

Application of the kinetic and isotherm models for better understanding of the mechanism of biomineralization process induced by Purpureocillium lilacinum Y3.

Purpureocillium lilacinum can promote the biomineralization of jarosite by secreting extracellular polymeric substances (EPS), but the detailed mechanism is not clear. In this study, the biosynthesis process of jarosite induced by P. lilacinum Y3 and hypha cell surface characterization were investigated. X-ray diffraction (XRD) analysis indicated that P. lilacinum Y3 could induce the formation of jarosite crystal and enhance mineralization kinetics. The kinetic and isotherm models confirmed that the metal ions transferring from the solution to the mycelium surface was controlled by diffusion process and the active interfacial sites on hypha cell surface played a pivotal role in the biomineralization process. Furthermore, transmission electron microscopy (TEM) pictures illustrated that the P. lilacinum Y3 mainly induced the generation of mineral precipitate extracellularly, but not intracellularly. Three-dimensional excitation-emission matrix (3D-EEM) fluorescence spectrum results further revealed the extracellular compounds such as fulvic-acid-like and protein-like substances participated in the mineralization process.

Trichosporon fermentans biomass flocculation from soybean oil refinery wastewater using bioflocculant produced from Paecilomyces sp. M2-1.

The soybean oil refinery (SOR) wastewater contains a high concentration of chemical oxygen demand (COD) and lipid, so the direct emissions of SOR wastewater will result in environmental pollution and waste of resources. Oleaginous yeast Trichosporon fermentans can consume organic materials in SOR wastewater to synthesize microbial oil, which achieves the purpose of SOR wastewater resource utilization. The effective harvesting technology of oleaginous yeasts can improve the utilization efficiency. In this study, Paecilomyces sp. M2-1 with high flocculating activity was isolated. The flocculants produced by M2-1 (MBF2-1) include 75% (w/w) polysaccharides, rely on cations, and display the flocculation percentage of above 77% in the range of pH 2-11. Especially under alkaline conditions, the flocculation percentage can be kept above 97%. The results of scanning electron microscope observation and zeta potential measurements suggested that the bridging, net trapping, and sweeping were the main flocculation mechanism of MBF2-1. MBF2-1 could flocculate T. fermentans that was used to reduce the organic matter in SOR wastewater and to produce microbial oil. Under the optimum conditions, the flocculation percentage of MBF2-1 against T. fermentans from SOR wastewater can reach 95%. Fatty acid content percent in microbial oil from T. fermentans was not almost affected by flocculation of MBF2-1. Moreover, MBF2-1 can further remove 55% and 53% of COD and oil content in the fermented SOR wastewater, respectively. The properties and high flocculating percentage displayed by MBF2-1 indicated its potential application prospect in oleaginous yeast harvest and food industry wastewater treatment.

Dibutyl succinate, produced by an insect-pathogenic fungus, Isaria javanica pf185, is a metabolite that controls of aphids and a fungal disease, anthracnose.

BACKGROUND: An entomopathogenic fungus, Isaria javanica pf185, causes mortality in nymphs of the green peach aphid and inhibits the growth of fungal plant pathogens. However, the metabolites of pf185 involved in these antifungal and aphicidal activities are unknown. This study was performed to identify the metabolites with these activities. RESULTS: An antifungal metabolite was purified by repetitive column chromatography and preparative high-performance liquid chromatography. Based on data from mass spectrometry and nuclear magnetic resonance, the active metabolite was identified as dibutyl succinate. The minimum concentration of dibutyl succinate to inhibit germination of conidia of the cause of anthracnose, Colletotrichum acutatum, was 4 mg mL-1 . Dibutyl succinate at 2 µg mL-1 inhibited mycelial growth. It also had strong aphicidal activity against the nymphs of the green peach aphid, Myzus persicae, median lethal concentration (LC50) of 306 mg L-1 at 24 h exposure, and median lethal time (LT50) of 14.5 h at 388 mg L-1 exposure. CONCLUSION: This is the first report indicating that a single metabolite, dibutyl succinate, from the beneficial fungus I. javanica, has potential for use in integrated pest management to inhibit both insect and fungal plant pathogens. © 2018 Society of Chemical Industry.

Anticancer Action and Mechanism of Ergosterol Peroxide from Paecilomyces cicadae Fermentation Broth.

Isaria cicadae, a medicinal food fungus, is a fruit from Paecilomyces cicadae. In this study, we purified ergosterol peroxide (EP) from the fermentation broth of P. cicadae and investigated its effects on renal cell carcinoma (RCC) cells, in vitro. EP was purified from P. cicadae fermentation broth. The human RCC cell line 786-0 was used to analyze the anticancer mechanism of EP and inhibit its effect on cancer cell proliferation, in vitro. EP with a validated structure showed a yield rate of 20.1 mg/L and a purity of 96%. EP significantly inhibited RCC cell growth and clone formation in vitro. In addition, EP suppressed the migration and invasion, triggered the apoptosis, and modulated the cell cycle of RCC cells, in a dose-dependent manner. It also downregulated ß-catenin expression. EP could be routinely produced through P. cicadae. It fights RCC cells in vitro through multiple mechanisms, including suppressing cell growth, colonization, migration, and invasion, arresting the cell cycle, attenuating ß-catenin pathways, and triggering apoptosis.

Endophytic fungus Paecilomyces formosus LHL10 produces sester-terpenoid YW3548 and cyclic peptide that inhibit urease and α-glucosidase enzyme activities.

Endophytic fungi have been used to obtain novel bioactive secondary metabolites with potential applications in medical and agricultural sectors, which can also act as lead targets for pharmaceutical and medicinal potential. In the present study, the endophytic fungus Paecilomyces formosus LHL10 isolated from the root of cucumber plant was tested for its enzyme inhibitory potential. The ethyl acetate (EtOAc) extract of LHL10 was screened for its inhibitory effect on acetylcholinesterase (AChE), α-glucosidase, urease, and anti-lipid peroxidation. The findings suggest that the EtOAc extract from LHL10 possesses significant inhibitory potential against urease and α-glucosidase. The EtOAc extract was thus, subjected to advanced column chromatographic techniques for the isolation of pure compounds. The structure elucidation was carried out through spectroscopic analysis and comparison with literature data, and these compounds were confirmed as known a sester-terpenoid (1) and a known cyclic peptide (2). The enzyme inhibition bioassay indicated that Compounds 1 and 2 exhibited remarkable inhibitory rate against α-glucosidase and urease, with an IC50 value of 61.80 ± 5.7, 75.68 ± 6.2 and 74.25 ± 4.3, 190.5 ± 10.31 µg/g, respectively. Thus, the current study concludes the enzyme inhibitory potential of endophyte LHL10 and provides the basis for further investigations of bioactive compounds, which could be used as potent drugs for enzyme inhibition.

Antioxidant Activity and Infrared Spectroscopy Analysis of Alcoholic Extracts Obtained from Paecilomyces hepiali (Ascomycetes).

This research studies the influence of substrate on the antioxidant activity of alcohol extracts of Paecilomyces hepiali. We used corn, rice, millet, and peas as substrates. Antioxidant activity was measured with the DPPH radical scavenging method. Concentrations of extracts (6.25, 3.12, 1.56, 0.78, and 0.39 mg/mL) were applied in all evaluations. Overall antioxidant activity was expressed as the concentration of substrate that decreased DPPH radical levels by 50% (IC50DPPH) for 7 methanol and 7 ethanol extracts. A comparison of IC50DPPH allowed us to conclude that the methanol extracts are more active in scavenging stable DPPH radicals than are the ethanol extracts. The substrate with antioxidant properties most suitable for cultivation of P. hepiali was rice supplemented with non-defatted soy flour. The extract most effective in scavenging stable radicals was the methanol extract of sample 4 (IC50DPPH = 2.33 mg/mL) cultivated on rice with nondefatted soy flour. The methanol extract of sample 7 cultivated on peas was less effective (IC50DPPH = 11.50 mg/mL). By crystallizing these extracts, we managed to obtain sufficient quantities of 6 samples in a solid state, for which infrared spectra were measured and confirmed the presence of amino acids in the extracts.