Fish processing waste is a secondary raw material that can be used as part of value-added product systems. Fish protein hydrolysates contain essential amino acids and biologically active peptides that can be included in food formulations. This article introduces an improved enzymatic hydrolysis technology for obtaining protein hydrolysates from secondary fish raw materials. The study featured Central-East Atlantic cod (Gadus morhua) and its processing waste, which included protein-containing backbones, muscle tissue, and fins. Five different enzymes (Pancreatin, Collagenase, Protozyme B, Alcalase 2.4 L FG, and Enzy-Mix U) were applied in a wide range of concentrations. The new technology involved one additional step to improve the sensory properties: protein hydrolysates were treated with an acetic acid solution. The alkaline dissolution stage was also optimized to increase the degree of protein solubility. The resulting hydrolysates underwent a chemical analysis and high-performance liquid chromatography. Other tests made it possible to reveal their amino acid composition and molecular weight distribution. The fish protein hydrolysates demonstrated a high protein content of 85–90% and a high amino acid ratio (110–190%) for virtually all essential amino acids. The product yield ranged from 22 to 55%. As the concentration increased from 1 to 8 g per 1 kg of raw material, the yield of protein hydrolysate and the degree of protein hydrolysis also increased. However, the amount of lowmolecular-weight peptides also grew, which spoilt the sensory profile (bitter taste, strong fishy smell). Only Protozyme proved suitable for the food industry: its concentration could be varied whereas the enzymolysis time could be shortened, if necessary. Pancreatin and Alcalase 2.4 L FG provided high-yield protein hydrolysates for microbiological use.
A more efficient extraction of bioactive plant substances is a major biotechnological and pharmaceutical challenge. The composite structure of wood fiber creates a major diffusion barrier to the extraction of valuable secondary metabolites. This research established some reliable correlations between the mechanical activation modes, the structural and physical profile of tree bark, and the bioactive extraction kinetics. The study involved aspen (Populus tremula L.) bark harvested in the Tomsk Region, Russia. The mechanical activation was performed in an AGO-2S planetary mill. At a centrifugal acceleration of 300 m/s2 for 5–10 min, the yield of water-alcohol fractions increased by 9.5–19%; when it was raised to 600 m/s2, the yield increased by 40–44%. The yield of extracted polyphenols reached 34.6% at 1,395 rpm (7–10 min) and 61.5% at 1,820 rpm. The highest yield of salicin occurred at 1,820 rpm for 1–3 min and was between 0.80 ± 0.01 and 1.00 ± 0.02% by weight. A three-minute dispersion of aspen bark at 1,395 rpm increased the share of the carbohydrate component D1080/D1460, alcohol D1140/D1460, carboxyl groups D1280/D1460, -C=C- D1600/D1460, and OH groups D3400/D1460 while the number of alkyl substituents D2920/D1460 and carbonyl groups D1720/D1460 decreased. The mechanical activation of raw material during one-minute grinding at 1,395 rpm raised the antioxidant content to 1.15 ± 0.02 mol/kg and led to the formation of second type inhibitors. When the reactor speed remained 1,820 rpm for 5 min, the increase reached 1.66 ± 0.03 mol/kg, with an induction period of 90 min. The identified patterns of mechanical pretreatment increased the profitability and sustainability of producing valuable bioactive substances from renewable plant raw materials.
Oxidative spoilage of fresh food products results in significant economic losses in the food industry. Active packaging with natural antioxidants is a promising alternative to synthetic preservatives: it extends shelf life and provides consumer safety. The growing regulatory restrictions on synthetic antioxidants necessitate the development of natural analogs with GRAS status, i.e., generally recognized as safe. This study compared the antioxidant activities of different leaf extracts from Armenian tree species for active packaging prospects. Leaves of Quercus robur, Quercus iberica, Salix alba, and Lycium barbarum were collected during the full growing season in the Republic of Armenia. Their antioxidant properties were assessed using the kinetic method of cumene oxidation, which involved total antioxidant content (f [InH]) and the reaction rate constant with peroxide radicals (k7). This method provided a comprehensive description of the antioxidant mechanisms. The extract yields ranged from 3.4 ± 0.3% (S. alba) to 6.8 ± 0.3% (Q. iberica). The highest antioxidant content belonged to Q. iberica (1.34×10–4 mol/L) while the highest reactivity belonged to the S. alba extract (k7 = 1.01×105 L/mol·s). In DPPH equivalents, the antioxidant activity was 3.5–13.4 μg/mL, which was similar to that of green tea (8–20 μg/mL) and superior to rosemary fractions. The lack of correlation between the antioxidant content and the activity was due to the structural differences in the ph enolic components and their reaction mechanisms. The ethyl acetate extracts of Armenian wood species demonstrated high antioxidant activity at a low cost of raw materials ($5–10/kg vs. $15–50/kg of commercial antioxidants), which makes them a cost-effective and environmentally sustainable alternative to imported antioxidants.
Food safety being a primary concern for food producers, viticulture and winemaking are subject to strict regulations and quality control procedures. Before a food additive can be incorporated into a food formulation, it must be investigated for stereoisomers and associated potential risks. Selecting a specific isomeric form of an optically active compound to be used as a food additive is a critical task. This review explores scientific publications and regulatory documentation regarding such stereoisomers of tartaric acid (2,3-dihydroxybutanedioic acid) as natural dextrorotatory L(+) isomer of plant origin, synthetic levorotatory D(–) and dextrorotatory L(+)- isomers, racemic mixture (DL form), and mesotartaric acid. The research specifically focuses on L(+)-tartaric acid (food additive E334), obtained from grapes and their derivatives, as well as D(–)- and L(+)-tartaric acids synthesized from hydrocarbon raw materials. In addition, the article reviews the methods for monitoring the properties of different forms of tartaric acid as well as the methods for evaluation of origin of L(+)-tartaric acid. Based on toxicological data and regulatory standards, L(+)-tartaric acid is preferred in winemaking due to its natural origin and metabolic compatibility. In contrast, the synthetic D(–)- and L(+)-tartaric acid form is poorly metabolized, which may pose potential health risks.
Probiotics are live microorganisms that confer documented health benefits upon the host. Among these, Lactobacilli remain the most popular probiotic cultures, largely due to their efficacy in preventing metabolic disorders. This article evaluates the probiotic potential of several Lactobacillus strains as key functional ingredients in metabolic dietary supplements and functional foods. The research featured Lactobacillus plantarum (B-5772, B-11264, B-3242), Lactobacillus fermentum (B-7574, B-7573), Lactobacillus acidophilus (B-194, B-2585, B-2900), and Lactobacillus casei (B-7951). The disc diffusion method revealed the degree of antibiotic resistance while the agar diffusion method made it possible to assess their antagonistic activity. The nutrient media were tested for carbohydrate sources to maximize the biomass accumulation in optical density units. The chromatographic analysis demonstrated the qualitative and quantitative composition of metabolites, i.e., B vitamins, organic acids, and amino acids. The biocompatibility of the strains was determined by co-cultivation. B-194 and B-7951 were resistant to streptomycin. B-7951 exhibited intermediate resistance to azithromycin while B-7573 and B-7951 were resistant to gentamicin, and B-3242 and B-7573 demonstrated a lack of susceptibility to streptomycin. B-11264 and B-7573 showed high antagonistic activity (≥23 mm) against Pseudomonas aeruginosa; B-7573 was effective against Bacillus cereus; B-11264, B-3242, and B-7951 resisted Enterococcus faecalis; B-11264, B-3242 and B-7573 showed good resistance to Klebsiella pneumoniae. The strains accumulated biomass better in the presence of lactulose. The chromatographic analysis showed that B-3242 produced vitamin B1 (25.98 ± 0.33 mg/g); B-7573 generated vitamin B3 (5.60 ± 0.03 mg/g); B-7573 yielded citric acid (52.6 ± 0.7 mg/L); B-2585 produced citric (58.0 ± 0.8 mg/L) and succinic (326.5 ± 3.2 mg/L) acids; B-7951 generated oxalic (17.1 ± 0.2 mg/L) and tartaric (17.1 ± 0.2 mg/L) acids. B-11264 produced tryptophan (16.2 ± 0.2 mg/g), glutamic acid (15.1 ± 0.2 mg/g), glycine (19.1 ± 0.2 mg/g), and proline (21.8 ± 0.3 mg/g). The strains demonstrated biocompatibility in co-cultivation. The strains involved in this research represent promising candidates for integration into probiotic nutraceuticals and functional food formulations.
Availability of micronutrients remains a major global concern. Fortified foods are inherently rich in vitamins and minerals; for instance, folic acid, or vitamin B9, is essential for preventing such health issues as neural tube defects in newborns. However, the efficacy of functional food depends on the bioavailability and solubility of the fortifying nutrients. This study focused on the solubility folic acid in aqueous solutions with citric, ascorbic, and succinic acids at various concentrations (3, 5, and 10%). The methods of UV spectroscopy and high-performance liquid chromatography (HPLC) were employed to analyze the solubility of folic acid, with the resulting data processed using standard statistical methods. The dissolution medium had a reliable impact on the solubility of folic acid. Selecting an appropriate dissolution medium proved crucial for enhancing the bioavailability of folic acid in food fortification. The 10% aqueous solution of aminoacetic acid (glycine) demonstrated the greatest potential under standard conditions. This solution achieved the highest levels of folic acid absorption and release, surpassing those for the aqueous solution of folic acid. The absorption curves suggested a uniform molecular distribution within the solution, as well as the stability of the dissolved form. The results can be used to optimize existing food fortification technologies, thereby helping to prevent folic acid deficiency.
As the current agricultural production capacity fails to meet the growing demand for meat products, the global market might soon face meat shortages. Cultured, or cultivated, meat is a prospective solution to protein security challenges. Its bioproduction relies on a three-dimensional scaffold that defines the structural and mechanical properties of the final product. The food science is seeking new scaffolding materials that would yield cultured meat with targeted sensory and textural properties. This article describes the physical and chemical properties of marine collagen to evaluate its biotechnological potential as raw material for cultured meat scaffolds. The study investigated collagen derived from jellyfish (Aurelia aurita) and zander skin (Sander lucioperca) obtained by acid extraction. The physicochemical characterization involved the Laemmli electrophoretic method, capillary electrophoresis, FTIR spectroscopy, MTT assay, and 3D bioprinting. The collagen consisted of two polypeptide chains (α and β) with molecular weights of 240 kDa (A. aurita) and 220 kDa (S. lucioperca). The samples exhibited an optimal isoelectric point, which supported cell culture growth and development. Hydroxyproline, glycine, and proline provided molecules with robust scaffolding properties. The marine collagen demonstrated biocompatibility but no cytotoxicity. The collagen samples from A. aurita biomass and S. lucioperca skin demonstrated good prospects as biomaterial for scaffolds in cultured meat production.
Sturgeon waste is a raw material base for industrial food production. Sturgeon heads are responsible for the majority of waste, and their rational processing will allow for sustainable utilization of fish resources. This study focused on the chemical composition and biological value of sturgeon head tissues. The research featured muscle tissue, heads, and processed products of Russian sturgeon (Acipenser gueldenstaedtii). While the chemical composition was studied by standard methods, the biological value of protein was calculated according to the FAO/WHO Food Committee methodology. A topography of various body sections demonstrated the presence of light and dark muscles. Fat tissue accumulated in the subcutaneous, cartilaginous, and abdominal zones, while connective tissue was registered in different body parts. The chemical analysis of flesh from head, body, and tail revealed 76.0–82.9% water contents, 14.2–18.5% protein, 2.4–4.9% fat, and 0.1–0.6% minerals. The average fat content was 2.8 times lower than in commercial sturgeon. Soft tissues of sturgeon head contained 18.6–20.4% protein and 12.2–16.2% fat. These data are similar in composition but differ in nutritional value. The biological value of proteins in the flesh from sturgeon head was 44.1%, which was 2.6% higher than that of soft tissues. Sturgeon head fat contained 45.8% monounsaturated fatty acids, 11.0% ω-3 polyunsaturated fatty acids, and 15.2% ω-6 polyunsaturated fatty acids. As reliable sources of valuable protein and fat, sturgeon head recyclables proved to be promising ingredients of multi-component foods.
As a staple food product, milk requires special standards of quality and safety at all production stages. As cross-regional competition in the dairy sector continues to escalate, understanding consumer behavior is now a critical priority for domestic market studies. This article describes the drivers that determine consumer behavior on regional dairy markets in Russia. The research featured milk and other dairy products produced in the Republic of Crimea, Russia. The empirical research relied on a two-stage consumer survey (n1 = 400; n2 = 300) using a random sample. The statistical approach employed descriptive and multivariate techniques, e.g., factor analysis with Varimax rotation and agglomerative hierarchical clustering (SPSS 23.0). The study revealed significant gender differences in the factors of choice. Men focused on the taste, price, fat content, and shelf life (47%) while women paid more attention to the composition, compliance with state standards, and local production (53%). The factor analysis made it possible to identify a two-stage model of consumer choice, where safety parameters dominated at the cognitive stage (R2 = 0.834), and image-assortment properties prevailed at the final stage (R2 = 0.794). Price proved to be a major quality indicator when choosing between brands. At the macro level, however, price served as a determinant of accessibility, as evidenced by a reliable negative correlation between price dynamics and consumption volumes in 2016–2023. The cluster analysis revealed three groups of brands based on the degree of perception: mass brands, intermediate brands, and niche brands. The differentiated regional marketing strategies based on consumption patterns can be used to support local brands and stimulate demand for local dairy products. The effect of sensory properties on loyalty to regional brands is a promising research direction.
Reliable measurements are critical for the food industry. The measurement uncertainty assessment is an obligatory procedure stipulated in modern standards, e. g., general requirements for the competence of testing and calibration laboratories (ISO/IEC 17025); Guide to the Expression of Uncertainty in Measurement (GUM), etc. This research applied the hydrometer and pycnometer techniques to the uncertainty associated with milk density tests. The tests employed both hydrometer and pycnometer methods. Measurement uncertainty was calculated using the simulationbased approach and uncertainty distribution laws. The hydrometer method revealed such uncertainty sources as thermometer calibration, hydrometer calibration, and repeatability. For the pycnometer method, they included balance calibration, balance resolution, weighing repeatability, pycnometer calibration, and repeatability. Both techniques were provided with mathematical models that incorporated input variables with assigned distribution laws, standard uncertainties for these variables, and the total relative standard uncertainties. The total standard uncertainty for milk density measurement was 31 kg/m3 for the hydrometer method and 21 kg/m3 for the pycnometer method. The expanded uncertainties were computed at a 95% confidence level with a coverage factor of two. The measurement uncertainty depended on the method and the instrumental errors. The hydrometer and pycnometer tests showed different measurement uncertainty, depending on the method and instruments employed. The pycnometer method had lower uncertainty and provided more reliable and accurate measurements. Selecting an optimal method and accurate calibration minimize uncertainty; however, additional factors require further research to adapt the models for industrial applications.
Publications on functional condensed dairy foods focus on the formulation without considering the intermolecular interaction between a particular sweetener and casein micelles. Consequently, the structuring processes remain largely understudied. Electron microscopy and rheological studies may provide important data about the structure and mechanics of this interaction. This article introduces a comparative structural analysis of a condensed dairy product with sugar and its low-calorie substitute allulose. The research included fresh and 14-month-old original prodct with sugar and its experimental samples where sucrose was substituted with 40 and 60% allulose. The structural analysis involved the methods of electron microscopy and rheology. The physicochemical parameters and sensory profile were studied using standard methods. The electron-microscopic studies of the structures formed by casein micelles showed that 40% allulose had no effect on their morphometry. However, replacing 60% sucrose with allulose destroyed casein micelles, encapsulating them with saccharide and forming dense aggregates, which spoiled the sensory and physicochemical quality of the final product. The excessive fluidity of the product was due to a 27% decrease in viscosity. In this study, replacing more than 40% of conventional sweetener with allulose damaged the quality of the functional condensed dairy product.
Barley (Hordeum vulgare L.) is a versatile crop utilized for food, feed, and industrial purposes; however, it remains highly sensitive to water stress. Chitosan-based biofertilizers reduce soil drought, thus improving the growth rate and yield of cereal crops. The article describes the effect of the chitosan biofertilizer Yunigel Plantum on barley resistance to soil drought. The study involved the Biom barley variety subjected to two triplicated series of experiments with Yunigel Plantum. The experiments featured the effect of Yunigel Plantum on seed germination energy under optimal conditions, as well on drought resistance. Yunigel Plantum improved the water status, reduced the oxidative damage, and stabilized the accumulation of such protective metabolites as anthocyanins and proline. The high levels of photosynthetic pigments reduced the negative impact of water stress on the photosynthetic system. Yunigel Plantum contained hydrogel structures that promoted water retention in the soil, making it more available to plants and reducing the stress. The synergy of these positive factors increased the drought resistance and reduced the damage caused by water deficiency. In this research, Yunigel Plantum showed good prospects for field trials in the sphere of barley farming.
Birch chaga is a popular functional component. This fungus is rich in chemical and biologically active substances, but the composition is so complex that their extraction remains a challenge. This article describes the functional and medicinal status of Inonotus obliquus Pil. in terms of its prospects for the food industry. The research featured the conk of I. obliquus, its in-vitro cell suspension culture, and callus cell culture. A set of standard research methods made it possible to study such aspects as microscopic parameters, chemical composition, and methods of introducing chaga into in-vitro culture, as well as the antioxidant and antimicrobial activity of chaga extract. I. obliquus proved to contain such beneficial elements as P0, Na+, K+, Ca2+, and Mg2+. The list of biologically active substances included melanin, vitamins C and E, selenium, β-carotene, etc. The main biological component was a polyphenoloxycarbonate complex, an aqueous extract that formed a colloidal polydisperse system. The rational conditions for I. obliquus suspension culture were as follows: cultivation for 30 days in the dark at 27 °C and 60–70% humidity. The optimal nutrient medium consisted of 40.0 glucose, 5.0 peptone, 2.0 yeast extract, 1.0 KH2PO4, 0.5 MgSO4, 8–9 agar, and 0.5 g/l potato starch. The highest growth index was 15.9 g. The antioxidant activity across the experimental aqueous extracts were similar, ranging from 27 to 31 kC/100 g. The highest antimicrobial effect was detected against Escherichia coli, Proteus mirabilis, Proteus vulgaris, and Pseudomonas aeruginosa. The experimental functional product from wheat flour and I. obliquus was reliably rich in beneficial bioactive substances, which rationalizes the current popularity of chaga fungus in the modern food industry.
Modern cheese-making trends require a specialized approach to domestic starter cultures adapted to specific cheese production parameters. Studying and systematizing the properties of collection strains of propionic acid bacteria will enable their use in specific technological schemes of production, thereby increasing stability and reducing the risk of defects. The aim of the study is to identify strains of propionic acid bacteria from the VNIIMS microorganism collection that possess industrially significant properties and are promising for the production of semi-hard cheeses with low-temperature second heating. Ten strains of propionic acid bacteria were investigated. Gas-forming activity was determined using graduated Dunbar vessels; antagonistic activity of the main and protective starter microbiotas against the strains was assessed by the well diffusion method; development in model nutrient media with varying pH levels, nitrate concentrations, lysozyme, and salt was measured spectrophotometrically at a wavelength of 600 nm. In cheeses, the dynamics of the development of the studied bacteria and the formation of organoleptic properties were evaluated using standardized methods. Based on gas-forming activity results, 6 promising strains of propionic acid bacteria were selected for further studies of technologically significant properties and cheese production trials. For active gas formation, the investigated strains require nitrogenous substances in an accessible form. Active acidity below 5.6 units was a critical factor for their development. Lysozyme at a dose of 2.5 g/100 L had no negative effect on the development of propionic acid bacteria, while nitrates at doses of 5–15 g/100 L reduced the growth rate of most selected strains in the majority of cases. Protective cultures Lacticaseibacillus rhamnosus, Lacticaseibacillus casei, Lacticaseibacillus paracasei exhibited selective antagonistic effects on some of the studied strains, while Lactiplantibacillus plantarum cultures inhibited their development. Antagonism toward propionic acid bacteria from the main starter microbiota – lactococci – is strain-specific. Cheese ripening with the studied bacteria under optimal temperature regimes in polymer packages selected for gas permeability influenced the development of the eye pattern in the cheese matrix, while the use of concentrated brine during salting contributed to reducing the number of eyes in the sub-cortical layer. Promising strains of propionic acid bacteria were selected based on their gas-forming ability and resistance to adverse environmental factors, which, in combination with targeted use of specific technological techniques, ensure cheese quality stability.
Advanced food production relies heavily on functional additives. Polysaccharides are functional food additives that combine the technological properties of thickeners, stabilizers, and emulsifiers. The absence of domestic cellulose and hemicellulose production remains a significant barrier to import substitution in Russia. This article introduces new processing technologies for producing commercial cellulose and hemicellulose from depectinized and delignified sugar beet pulp. The experiment featured the effect of such variables as sodium hydroxide concentration and processing time on the efficiency of hemicellulose extraction. The obtained cellulose and hemicellulose were tested for the effect of washing on purity, as well as the effect of hydrochloric acid treatment on cellulose yield. The sensory and physicochemical tests involved standard research methods. The most effective production conditions were as follows. Hemicellulose was extracted with a 2% sodium hydroxide solution at 25°C for 3 h and washed with 70% ethanol. Cellulose was treated with a 15% hydrochloric acid solution for at 60°C 5 h. The resulting cellulose and hemicellulose met the sensory and physicochemical standards in all aspects but raw material. Beet pulp proved to be a promising source of cellulose and hemicellulose with good import substitution and recycling prospects. However, the use of beet cellulose and hemicellulose in the food industry requires updating the existing regulatory documentation.
Utilization and processing of milk whey have become a pressing problem for the dairy industry as a result of unprofitable production methods and the poor energy-saving industrial technologies. Milk whey is a valuable raw material that contains more than 200 organic and mineral components. It can be processes into products with useful biological properties. The article describes a new fortified protein curd whey product obtained with magnesium thermal coagulation of proteins and fermented with a complex lactobacillus starter. The curd whey was obtained by acid-rennet coagulation as a by-product of cottage cheese with a fat mass fraction of 9%. After +35–40°C in water bath, fat and casein powder were removed in a cream separator. The permeate was obtained by thermal denaturation at 92 ± 1°C with a 10% MgCl2×6H2O solution in concentrations of 2–42 g/L. The protein was then subjected to vacuum filtration. The fermentation of whey proteins involved a complex starter culture of four Lactobacillus strains. The research relied on standard physicochemical and microbiological methods. The optimal mode for thermal magnesium denaturation of milk whey was 92 ± 1°C for 5–10 min with 18 g/L MgCl2. It yielded 25.0 g/L protein with 2.4 mg/g magnesium. When the magnesium concentrate of thermally precipitated whey proteins was fermented with the complex starter culture, it activated the proteolysis process, as well as provided the targeted physicochemical and sensory properties. After freeze-drying at –40°C for 18–20 h, the powder obtained was rich in living lactobacilli cells (1–3×107) and contained 907 mg/100 g magnesium. The new method made it possible to turn whey wastes into a magnesium-containing concentrate of thermally precipitated proteins with increased biological value. Its qualitative indicators provide a wide range of possibilities in various areas of the food industry.
Omics technologies give consumers access to personalized disease prevention. Today, commercial genetic testing is very popular. For instance, nutrigenetic testing requires no medical consultation or prescription. Personalized nutrition is an informed diet choice based on genetic, phenotypic, climatic, geographical, and environmental data. It is a more effective preventive measure than standard dietary recommendations. This article describes the state and prospects of consumer genomics with an emphasis on nutrigenetic testing as a basis for personalized dietary choice. The review covered scientific publications registered in PubMed, ScienceDirect, and eLIBRARY.RU in 2012/2020–2025, as well as popular science literature and marketing reports. The development of human genomics had passed through several stages before it gave start to personalized nutrition as part of consumer genomics. Its popularization on the Russian nutrition market has its own peculiarities. The topic field includes such aspects as nutrigenetics, nutrigenomics, genome-wide association studies, polygenic risk index, epigenetics, and metabolomics of microbiome. The bioinformatics and machine learning help to process and interpret personalized data. Genetic risks awareness increases the motivation for healthier lifestyle choices. While the field of genetic research and personalized nutrition is run by some foreign and Russian direct-to-consumer companies, legal regulation remains a vague issue in this sphere. Personalized nutrition requires a competent use of interdisciplinary achievements combined with ethical standards and regulatory norms. However, serious preclinical and clinical studies are needed to introduce personalized nutrition into the state healthcare system.
New breeds of fur-bearing animals are multi-breed crosses. Selection requires new methods for genetic control. This article describes a new early-maturing and highly-productive crossbreed of rabbits called Rodnik obtained at the Afanasyev Research Institute for Fur Farming and Rabbit Breeding, Russia. It was a result of multi-stage generations of crossings between such breeds as White Giant, Soviet Chinchilla, and Californian. However, in terms of polylocus genotypes of highly polymorphic elements, the Rodnik rabbits are closer to the White Giant breed. The article describes the changes in the genetic pool of this three-breed cross through a comparative analysis of the nucleotide sequences of exons 8 and 13 of the melanophilin (mlph) color gene. The research featured the rabbit breeds of White Giant, Soviet Chinchilla, and Californian, as well as their Rodnik cross. The analysis of the mlph gene in Oryctolagus cuniculus involved bioinformatics methods based on the nucleotide sequence of the reference genome UM_NZW_1.0 (GCF_009806435.1) from the GenBank database. The exon-intron structure was determined using the Splign software; the conditions and stages of PCR, including the design of primers for exonic regions, were developed using the online version of BLAST3. Exons 8 and 13 made it possible to identify the differences between chinchilla-colored rabbits and other breeds. In some color properties, the Rodnik cross was closer to the Californian parent breed. A single nucleotide polymorphism (SNP) was detected in two nucleotides in exon 8 and in one nucleotide in exon 13. The haplotypes of two SNPs in exon 8 and one SNP in exon 13 of the mlph gene were homozygous and coincided with the White Giant breed. The Soviet Chinchilla and Californian breeds carried a heterozygous haplotype for two SNPs in exon 8, as well as in the homozygote of another nucleotide (C–T) in exon 13 in the same positions. The identified SNPs did not correlate with color. However, the Rodnik cross was closer to the White Giant in terms of genotypes and polymorphisms of some highly polymorphic genomic elements. The mlph gene might become part of selection as it belongs to the Ras oncogene superfamily and the largest exophilin subfamily of Rab effector proteins that coordinate vesicular transport and some adipogenesis stages.
Extraction is the initial stage in the study of the chemical composition of plant material. Crude extracts contain bioactive components that can be used in pharmaceuticals, food products, and food additives. Advanced extraction methods produce more yield and consume less time. The choice of a perfect extraction method depends on the target substance, the type of plant material, and its cell structure. Optimal extraction conditions are determined experimentally for each plant species. In this research, we revealed the main extraction factors for cell cultures of medicinal plants, depending on the technological limitations caused by the internal structure of the raw material. The bioactive extraction procedure was optimized using the experimental planning method. The study featured cell cultures of Ginkgo biloba L., Pulmonaria officinalis L., Filipendula ulmaria L., and Scutellaria baicalensis Georgi. The variables to be studied during the extraction experiments included ethyl concentration, temperature, and time. All the extracts were tested for optical density using spectrophotometry, with a full factorial experiment with three variable parameters. The data obtained underwent the analysis of variance (ANOVA) to check the mathematical model that defined the correlation between the flavonoid yield and the main extraction parameters. Using the experimental planning method, we optimized the main factors that facilitated the extraction of bioactive compounds from medicinal plant cell cultures. The volume fraction of the solvent proved to be associated with the largest yield of bioactive compounds from the plant cell cultures. Its optimal amount was the same across the samples (70% ethyl alcohol). The optimal extraction time and temperature for callus cultures of Filipendula ulmaria L. and Scutellaria baicalensis Georgi were 5 h and 35°C, respectively. The callus cultures of Pulmonaria officinalis L. demonstrated the most economically advantageous extraction parameters, i.e., 2 h and 30°C. The optimal extraction parameters for Ginkgo biloba L. were 6 h and 55°C. The study revealed the optimal parameters for the extraction of bioactive compounds from medicinal plant cell cultures with the maximal flavonoid yield. The results be used in further research.
Plant raw materials are an accessible and efficient substrate for multipurpose enzymes and multi-enzyme complexes obtaining. Thermophilic bacteria are reliable producer strains for industrial enzymes with valuable physicochemical properties, e.g., heat resistance, stability at extreme pH, and chemical stability. Food scientists are on the lookout for new thermophilic strains capable of producing high yields of industrially valuable enzymes from cheap plant raw materials. In this research, various plant cultivation media affected the complex hydrolytic enzymes produced by a thermophilic strain of Bacillus subtilis Kb.12.Gl.35. The research objective was to select the optimal raw materials for the synthesis of thermostable hydrolases, i.e., proteases, amylases, and hemicellulases. The thermophilic strain was isolated from a compost sample, identified by 16S rRNA sequencing, and tested using standard microbiological and biochemical methods. The strain grew on different media with plant flour combined with yeast or corn extract. The activity of extracellular enzymes made it possible to study the composition of the hydrolase complex. The proteolytic activity was determined using casein as the substrate and by zymography. The 3,5-dinitrosalicylic acid (DNS) method revealed the carbohydrase activity. When culturing the hydrolase producer on nutrient media with plant flour (1%) and corn extract (0.5%), the highest level of protease synthesis (≥ 40 units/mL) was detected in the samples with amaranth, oat, and rice flour. The highest level of amylase (≥ 1,300 units/mL) belonged to the chickpea, oat, and rice flour samples. The biggest yields of galactomannanase (≥ 200 units/mL) and xylanases (≥ 60 units/mL) was found in the samples with amaranth, pea, and chickpea samples. The biggest amount of arabinogalactanase (≥ 35 units/mL) belonged to the amaranth, chickpea, and rice flour. Based on the zymographic analysis, the enzymes of 24.7–28.2 kDa and the proteases of 62.7–75.0 kDa appeared to be the most efficient proteolytic agents. Their activity was obvious on plant raw materials, in contrast to the standard LB medium. The secretion level of 15.1 kDa proteases was the same on the plant media as on the standard medium. Bacillus subtilis Kb.12.Gl.35 proved to be an effective producer of proteases, amylases, and hemicellulases with plant flour and corn extract in the medium. Further studies are needed to optimize the composition of nutrient media and cultivation conditions to increase the yield of the target hydrolytic enzymes.
