Plant cells and tissue cultures are sources of secondary plant metabolites. Substances produced by callus cultures can expand the raw material base in pharmacy and food production. However, isolating biologically active substances from medicinal plants is a labor- and time-consuming process. As a result, new and efficient technological processes adapted for extraction from callus cultures are in high demand, and new algorithms of isolation and purification of biologically active substances remain a relevant task. This research featured callus cultures of Scutellaria baicalensis. The procedures for phytochemical analysis and isolation of biologically active substances involved such physicochemical research methods as high-performance chromatography (HPLC), thin-layer chromatography (TLC), UV spectrometry, and IR spectrometry. The high performance liquid chromatography confirmed the presence of flavonoids represented by baicalein (5,6,7-trioxyflavone), baicalin (baicalein 7-O-glucuronide), scutellarein (5,6,7,4-tetraoxyflavone), scutellarin (7-O-glucuronide scutellarein), vagonin, and oroxylin. The spectral analyses also detected skutebaicalin. The highest total content of diterpene belonged to the samples extracted with 70% ethanol at 70°C. The content of diterpene was 0.09 mg/cm3 in terms of betulin. The biologically active substances were isolated from the callus extracts of S. baicalensis with a recovery rate of ≥ 80%. The purification scheme made it possible to obtain highly-pure individual biologically active compounds: trans-cinnamic acid, baicalin, and oroxylin A had a purity of ≥ 95%; baicalein had a purity of ≥ 97%; scutellarin and luteolin reached ≥ 96%. The new technological extraction method made it possible to obtain extracts from S. baicalensis callus cultures, which were tested for the component composition. The developed isolation algorithm and purification scheme yielded biologically active substances with a purification degree of ≥ 95%.
This paper aimed to study the biodegradation of collagen-containing waste (pork skin) induced by collagenase and Neutrase 1.5 MG enzymes and compare the amino acid, peptide, and carbohydrate composition of hydrolysis products. It was found that the degree of biodegradation of collagen-containing raw materials (pork skin) reached 78% when using an enzyme preparation (collagenase with a concentration of 250 U/g of the substrate) at pH 7.0, 40 °C, and a 360 min process duration. It was shown that the content of peptides with a molecular weight of 6.5–14.0 kDa in the hydrolysis products (collagenase) of collagen-containing wastes was 13.4 ± 0.40%, while in the products of hydrolysis (Neutrase 1.5 MG) it was 12.8 ± 0.38%. The study found that the hydrolysis products (Neutrase 1.5 MG) of collagen-containing raw materials contain fewer hexoses, free hexosamines, and hyaluronic acid than the hydrolysis products (collagenase) of collagen-containing raw materials. The content of chondroitin sulfates is practically the same in all samples of hydrolysis products. Proteases with collagenolytic activity are widely used in industry. Recently, they have increasingly been used in pharmaceutical, food, and other industries. Collagenases are promising enzymes for the production of chondroprotectors used for the treatment of osteoarthritis.
Bioactive peptides derived from food proteins are becoming increasingly popular ingredients due to their beneficial effect on the immune system and other functional properties. We aimed to develop a technology for obtaining peptides from poultry by-products and identify their bioactivity. Pepsin was the main reagent for the in vitro enzymatic hydrolysis. Specialized equipment and methods were used to determine the key indicators. The molecular weight and bioactivity of the resulting peptides were calculated by using the Peptide Mass Calculator and PeptideRanker online resources. First, we developed a flow chart for obtaining bioactive peptides and produced hydrolysates from poultry by-products. The hydrolysates had identical physicochemical parameters, with no significant differences. The molecular weight distribution revealed that most protein fractions were represented by peptides with a molecular weight below 20 kDa. Then, we evaluated the bioactivity of the peptides. The hydrolysate obtained using pepsin with an activity of 30 units per 100 g of material showed higher bioactivity in the FD peptides (0.922094). The hydrolysate obtained using pepsin with an activity of 45 units per 100 g of material had greater bioactive properties in the CYG p eptides (0.947378). Based on the results, we designed a flow chart for obtaining hydrolysates from poultry by-products and evaluated the bioactive properties of the peptides obtained. For further work, these properties should be confirmed by in vitro experiments to determine the reliability of our data and identify specific bioactive properties of the peptides.
Introduction. Cellulose-containing parts of herbs are an excellent source of alternative energy and can be used to produce biological ethanol. The present research aims at improving this fundamental and promising area of biotechnology. It introduces a new consortium of microorganisms that can saccharify while fermenting the substrate. Study objects and methods. The research featured technical cellulose obtained from Miscanthus sinensis using hydrotropic delignification and oxidation with pertrifluoroacetic acid. The ethanol content in the culture liquid was determined using an Agilent 7890B gas chromatograph with a flame ionization detector. The biocompatibility of the strains was studied by growing a direct co-culture in a dense nutrient medium. Results and discussion. The research objective was to create a new microbial consortium for the single-step production of bioethanol from Miscanthus sinensis cellulose. A set of biocompatibility experiments and cultivation conditions made it possible to select the optimal producers. The two developed microbial consortia required optimal compositions of culture media, which were determined by varying the ratio of components and measuring the yield of ethanol in the resulting culture liquid. Conclusion. The best consortium for Miscanthus sinensis cellulose consisted of Pichia stipites Y7124, Candida shehatae NCL3501, Kluyveromyces marxianus Y-4290, and Zymomonas mobilis 113 at a ratio of 1:1:1:1. The optimal parameters of bioethanol production included: temperature = 35 ± 1°C, pH = 5.2, time = 16 ± 1 h. The most efficient culture medium had the following composition (g/l): glucose – 5.0; peptone – 5.0; yeast extract – 0.4; K2HPO4 – 1.5; (NH)2 HPO4 – 1.5; MgSO4 – 0.5.
The work reveals the results of studying the content of biologically active substances in samples of extracts of Ginkgo biloba callus cultures. Callus cultures grown in vitro on liquid nutrient media were the objects of the study. Considering various factors affecting the yield of the target components during extraction, the volume fraction of the organic modifier in the extracting mixture, the temperature factor, and the exposure time were identified as the main ones. The maximum yield of extractive substances (target biologically active substances with a degree of extraction of at least 50%) from the samples of callus culture extracts was detected at a ratio of extragent of 70% ethanol, a temperature of 50 °C, and exposure time of 6 h. Flavonoids, such as luteolin, quercetin, isoramentin, kaempferol, and amentoflavone, were isolated in the extract samples. As a result of column chromatography, fractions of individual biologically active substances (bilobalide, ginkgolide A, B, and C) were determined. The proposed schemes are focused on preserving the nativity while ensuring maximum purification from associated (ballast) components. Sorbents (Sephadex LH-20, poly-amide, silica gel) were used in successive stages of chromatography with rechromatography. The degree of purity of individually isolated substances was at least 95%.
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