The generic term “melanin“ describes a black pigment of biological origin, although some melanins can be brown or even yellow. The pigment is characterized as a heterogenic polymer of phenolic or indolic nature, and the classification of eu-, pheo- and allo- melanin is broadly accepted. This classification is based on the chemical composition of the monomer subunit structure of the pigment. Due to the high heterogeneity of melanins, their analytical characterization can be a challenging task. In the present work, we synthesized the current information about the analytical methods which can be applied in melanin analysis workflow, from extraction and purification to high-throughput methods, such as matrix-assisted laser desorption/ionization mass-spectrometry or pyrolysis gas chromatography. Our thorough comparative evaluation of analytical data published so far on melanin analysis has proven to be a difficult task in terms of finding equivalent results, even when the same matrix was used. Moreover, we emphasize the importance of prior knowledge of melanin types and properties in order to select a valid experimental design using analytical methods that are able to deliver reliable results and draw consistent conclusions.

The ability to orally administer silver nanoparticles (AgNPs) in enteric capsules implies a direct interaction with the colon microbiota. The in vitro effect provides a portrayal of the functional properties under in vivo conditions. The purpose of this study was to describe a green AgNP synthesis process, using aqueous extract from Lactarius piperatus mushroom, and to characterize the nanomaterial. We determined its antimicrobial and antioxidant effects in vitro in the microbiota of healthy individuals via the GIS1 system—a colon transit simulator. Per the quantitative polymerase chain reaction (qPCR) results, the antimicrobial properties of the AgNPs affected the initial share of different enteric species by decreasing the Bacteroides, Enterobacteriaceae, and Lactobacillus populations and favoring the Bifidobacterium group. The association between AgNPs and wild mushroom L. piperatus extract had a synergistic antibacterial activity against various pathogenic microorganisms while the mushroom extract reduced biofilm formation. Administration of AgNP maintained its constant antioxidant status, and it was correlated with a reduction in ammonium compounds. The physicochemical characterization of these NPs complemented their biochemical characterization. The maximum ultraviolet-visible spectroscopy (UV-VIS) absorbance was observed at 440 nm, while the Fourier transform infrared (FT-IR) spectrum reached a peak at 3296 cm^–1, which was correlated with the high-performance liquid chromatographic analysis (HPLC). The major phenolic compound was homogentisic acid. The size (49 ± 16 nm in diameter) and spherical shape of the NPs were correlated with their biological effects in vitro.

Environmental changes generate free radicals and reactive oxygen species (ROS), resulting in abiotic stress in plants and fungi. Gamma ionizing radiation generates a significant amount of free radicals and ROS, thereby simulating natural environmental stressors. We used a ⁶⁰Co source of radiation to experimentally induce oxidative stress in living mycelium mass of the medicinal fungus Fomes fomentarius, in order to obtain a late response of stress tolerance by means of bioactive compounds synthesis. We measured the response at 24, 48, and 72 h after the irradiation. The highest improvement was found 24 h after exposure for antioxidant activity and for total phenolic compounds of methanolic extract, with a 1.89- and 1.64-fold increase, respectively. The total flavonoids in methanolic extract increased 1.68 times after 48 h from treatment and presented a more stable raising in the assessed time-lapse. For the three analyzed parameters, 300 Gy was the optimum absorbed dose to trigger a beneficial response, with potentially applications in pharmaceutics and nutraceutics. Gamma irradiation can be used as a biotechnological tool to direct the secondary metabolites synthesis upregulation in medicinal mushroom living mycelium.

Submerged culturing of mycelium is an efficient technique used to increase biomass yields, more so when employed with naturally slow-growing species of mushrooms. This paper is concerned with optimizing nutrient broth components used in Inonotus obliquus cultures for achieving high biomass yields. We modeled the effect of seven biotechnological parameters (six broth ingredients and the initial pH of nutritive broth) on mycelial biomass and predicted an optimum broth formula using response surface methodology. An analysis of variance showed that the elaborated model is significant (F-value of 2.76 and p-value of 0.0316). We used bioreactor cultures to confirm the model’s optimum prediction and to compare these results with a general-purpose mycology medium, namely potato dextrose broth (PDB). The optimized bioreactor culture yielded 4.37 g/L (93.36% of the dry weight prediction), while the PDB bioreactor culture yielded 2.084 g/L, after 15 days of cultivation. The optimized formula was: 2.15299 g malt extract, 3.99296 g yeast extract, 11.0041 g fructose, 17.4 g soluble starch, 0.1 g MgSO₄, and 0.05 g CaCl₂ per liter of broth.

Inonotus obliquus is a parasite on the birch and other trees and is also a well-known medicinal mushroom. Its sterile conk is highly sought for its bioactive compounds such as phenols, polysaccharides, triterpenoids, and steroids. It was traditionally used to treat various gastrointestinal diseases, viral and parasitic infections, to counteract the progression of cancers, and to stimulate the immune system. We used acute gamma irradiation, followed by short-term submerged cultivation, as an oxidative stress inducer to enhance the synthesis of mycelial metabolites. The 300 Gy and 400 Gy doses showed the best results across the whole experimental design. Each assayed criterion had a different corresponding optimal stimulation dose. In one experiment, sublethal doses of irradiation triggered the dry weight of the cultured mycelium to increase by 19.764%. The free radical scavenging potential of the mycelium extracts increased by 79.83%. The total phenolic content of mycelium extracts and culture broth increased by 55.7% and 62.987%, respectively. The total flavonoid and sinapinic acid content of the broth increased by 934.678% and 590.395%, respectively. As such, gamma irradiation pre-treatment of the mycelial inoculum proved an interesting, economically and environmentally effective tool for stimulating secondary metabolite synthesis in submerged mycelium cultures.