[Alternative protein sources: bacteria Methylococcus capsulatus concentrate, characteristic of composition and biological value].

A promising growth vector of food protein production in the context of the Russian Federation's food sovereignty security is the use of microbial synthesis. Taking into consideration the proven promising use of biotechnological processes in the production of alternative protein sources, modern scientific research is focused, among other issues, on improving the technology of obtaining food microbial protein using a variety of substrates and strains-producers, as well as evaluating the consumer properties, food, biological value and safety of such products. The purpose of the research was to study and comparatively evaluate protein concentrate (PC) from bacteria Methylococcus capsulatus and basic food of animal and plant origin within the development of the technology of optimal in nutritional and biological value PC production. Material and methods. Analysis of the nutritional and biological value of PC obtained from denucleinized and purified from cell walls biomass of methanoxidizing bacteria Methylococcus capsulatus (strain GSB-15) was carried out on 46 indicators, including estimation of protein content and amino acid composition, fat content and fatty acid composition, ash and moisture. Biological studies based on measuring of net protein ratio / net protein utilization were performed on 28 growing (between 25-50 days of life) male Wistar rats. Rats in the control group (n=14) received a semi-synthetic casein diet with a protein content of ~12% in calories, the test group (n=14) received a diet including an equivalent amount of PC protein. Body weight, feed intake, and fecal and urine nitrogen losses were measured during the experiment. The biological value and digestibility of protein were judged by coefficients of protein efficiency ratio, net protein ratio, true protein digestibility, true protein biological value, true net protein utilization. Results. The nutritional value study of PC showed high protein content - 69.0%, the share of fat, moisture and ash, accounted for 0.17, 9.5 and 14.4%, respectively. The carbohydrate content was 7.0% (of which mono- and disaccharides were <0.1%). The results of a comparative assessment of Methylococcus capsulatus protein amino acid profile and basic food of animal and plant origin showed a balanced content of the most amino acids, the level of which is comparable with the protein of chicken egg, which is traditionally a standard of quality of complete protein. At the same time, the content of the essential amino acid tryptophan in PC was an order of magnitude lower than in chicken egg protein; the content of this amino acid in PC is comparable with incomplete plant proteins (sunflower, flax, rapeseed). The results of the biological value evaluation of the Methylococcus capsulatus protein in the experiment on rats indicate a relatively low biological value of the microbial synthesis protein, that is caused, most likely, by tryptophan deficiency. Rats of the test group had a significant decrease in body weight gain, feed/protein intake, coefficient of protein efficiency ratio, coefficient of net protein ratio, true protein biological value, true net protein utilization. Conclusion. The results of a comparative evaluation of PC from methanotrophic bacteria Methylococcus capsulatus denucleinized biomass and basic food of animal and plant origin indicate its relatively high nutritional value. However, the characteristics of this PC sample were not optimal in regard of protein biological value by reason of tryptophan deficiency. A single amino acid deficiency is not a valid argument for not using microbially synthesized protein in human nutrition, considering the capabilities of the modern food industry, including ways to enrich foodstuffs with missing components. In addition, there is every cause to believe that adjusting the hydrolysis technology used in the production of PC will allow to eliminate the essential amino acid loss, thereby increasing the biological value of this product.

[Squid fat as a promising source of polyunsaturated fatty acids].

Omega-3 polyunsaturated fatty acids (ω-3 PUFA) are substances that play an important role in human metabolism. They are essential nutritional factors and can improve the functioning of individual systems and the body as a whole. The main source of ω-3 PUFA has long been fish fat, which contains PUFA in the triglyceride form. A fairly new and promising alternative to fish fat is the liver fat of the Commander squid (Berryteuthis magister), which additionally contains alkylglycerols, contains PUFA in the phospholipid form and can be obtained from squid fishery waste. The objective of the research was to carry out an analysis of scientific data, including the results of studies of the biological activity of squid fat, as well as its components that are part of other similar raw materials, and evaluate the prospects for its use in medical practice. Material and methods. During the study, various sources were analyzed, including scientific literature from electronic databases eLibrary, PubMed, Scopus, Web of Science and electronic search engines Google Academy from 2000 to 2022. Results. It is noted that squid fat has a pronounced biological activity. Its components increase innate immunity, have antitumor potential, improve the state of the body under stress, have hypolipidemiс and hypotensive effect, improve memory and attention, and also positively affect the composition and rheological parameters of blood. In addition to these effects, a positive effect of ω-3 PUFA and alkylglycerols on spermatogenesis, sperm quality and the female reproductive system has been noted. In a number of studies, alkylglycerol esters increased the permeability of the blood-brain barrier and, due to their structure, are able to form vesicles, therefore, they can be considered as raw materials for the production of new dosage forms for targeted therapy of brain tumors. In the available literature, in the case of the use of squid fat and its components, undesirable side effects have not been identified. Conclusion. Squid fat is a complete source of ω-3 PUFA and alkylglycerols, therefore it can be recommended as a dietary supplement, especially in a diet low in ω-3 PUFA.

[Ways of nutritional value increasing of distillary grain fiber].

In the process of grain processing for ethyl alcohol, practically only carbohydrates are consumed, which are presented mainly in the form of starch. The remaining components (protein, fats, fiber, minerals) in transit pass into the grain fiber remaining after distillation of the alcohol from the mash. Distillery grain fiber surpasses wheat bran in its indicators, since during the processing it is enriched with biomass of alcohol yeast. In addition, there is a technological possibility of its additional enrichment with protein, amino acids, and vitamins due to changes in the modes of alcoholic fermentation. The aim of the work was to assess the influence of the mode and conditions of alcoholic fermentation on the composition of distillery grain fiber. Material and methods. Under laboratory conditions, samples of grain fiber of alcohol production were obtained by the method of fermentation samples from wheat under various fermentation conditions. In the obtained samples, the following parameters were determined: protein according to Barnstein, crude protein, vitamins B1, B2, B6, E, as well as the amino acid composition. The volume fraction of ethyl alcohol, the mass concentration of fermentable carbohydrates, and the concentration of yeast cells were determined in intermediate products of alcohol production. Results and discussion. Studies on the effect of yeast from various manufacturers on fermentation rates, biomass growth and grain fiber composition showed the advantage of race Y-717 in terms of alcohol accumulation (11.5% vol.), Fermentation rate (56 hours) and yeast cell concentration (260 million/cm3), which was 15-30% more than in other options. However, from the point of view of increasing the content of protein according to Barnstein and crude protein in samples of grain fiber with yeast Y-717, an increase of only 3-4% was noted compared with other options. This was associated with a decrease in the concentration of yeast by 72 h of fermentation due to autolysis. Studies on the influence of the initial yeast concentration on the growth of biomass, the fermentation rate and grain fiber indices showed that with an increase in the initial yeast concentration from 15 to 45 million/cm3, the fermentation time reduced to 48 h, the biomass growth at the end of fermentation was 20%, the protein in grain fiber increased by 15%, the content of vitamins B1, B2, B6 and E as well as amino acids increased by 13-17%. Conclusion. According to the results of the studies, the technological possibility of enriching distillery grain fiber with protein, amino acids, and vitamins due to a change in the fermentation process during the processing of grain raw materials to alcohol is shown. In particular, this could be achieved through the use of yeast with a high growth rate, by increasing the concentration of yeast biomass, shortening the fermentation period and preventing yeast autolysis at the maturation stage.

[Databases of the chemical composition of foods in the era of digital nutrition science].

By studying the chemical composition of foods, expanding the list of data on the content of nutrients, including minor biologically active substances, in the era of digital nutrition science, it became possible to create relevant systematic databases of the chemical composition of foods and rations in general. They allow us to solve various problems of modern society from the point of view of nutrition science. This review aim to analyze and generalize modern approaches to the formation and updating of databases of the chemical composition of food products from the standpoint of digital nutrition science. Results. This review considers the main provisions regarding creation of databases, directions for the development of food chemistry, discusses existing international programs for collecting and compiling data. The methods of systematizing data on the qualitative composition and content of biologically active and minor substances in products, as well as the problems associated with the development and metrological certification of highly selective highly sensitive analytical methods necessary to obtain reliable and reproducible data are considered. Conclusion. The development of digital nutrition science significantly increases the availability and quality of information on the chemical composition of foods, and allows it to be updated quickly. Further improvement of the quality of the data presented in the tables of chemical composition is associated with the establishment of stability and relationships between micro- and macro-components, their influence on the safety, stability of the chemical structure, the influence of the physic-chemical characteristics of the matrix on nutritive value of foods, determination of the content of specific minor components, development of relevant regulatory documents.

[Biological value of fruits and berries of Russian production].

Fruits and berries are the most important sources of a wide range of biologically active substances, including vitamin C, carotenoids, flavonoids, anthocyanins. In order to replenish and update data on the content of food and biologically active substances in the tables of the chemical composition of food products, a study of the content of mono- and disaccharides, dietary fiber was carried out; vitamins C, B1, B2 and E, minerals and trace elements; flavonoids (in terms of rutin), anthocyanins; organic and hydroxycinnamic acids, stilbenoids in various varieties of 16 fruit and berry crops. Material and methods. The material for the study was the fruits of promising varieties and selected forms of pome fruit (apple, pear), stone fruit (cherry, plum, apricot), berry (garden strawberry, raspberry, black currant, red currant, gooseberry), non-traditional crops (actinidia, honeysuckle, cornelian cherry, viburnum, sea buckthorn, rosehip) - a total of 208 samples grown at the I.V. Michurin Federal Scientific Center. The B vitamins were determined by the fluorometric method, and vitamin E, organic and hydroxycinnamic acids, carbohydrates, and stilbenoids were determined by HPLC. The amount of anthocyanin pigments was determined by pH differential spectrophotometry, dietary fiber - by enzymatic-gravimetric method, flavonoids - spectrophotometrically. Results and discussion. The main carbohydrate of apricot is sucrose, black currants, cherries, raspberries and honeysuckle contain mainly fructose and glucose. Raspberries and currants are high in fiber. Comparison of the obtained data on the content of vitamins B1, B2, E, flavonoids and anthocyanins in the studied population of varieties in comparison with the published data of tables of the chemical composition of food products in the USA and Russia was carried out. By the content of vitamin C in descending order, the fruits are arranged in a row: black currant > sea buckthorn > honeysuckle > strawberry > red currant > viburnum > gooseberry > raspberry > apple. Berries, making a significant contribution to providing the body with vitamin C, are not an essential source of vitamins B and E. The inclusion of 100 g of fresh fruit in the diet provides about 10% of the recommended dietary intake for potassium (apricot, gooseberry, cherry and black currant), magnesium (apple, cherry, strawberry) and dietary fiber. Honeysuckle and black currant are high in anthocyanins; consumption of 100 g of these berries will ensure adequate intake of these micronutrients. Some varieties of strawberries, apples and pears are rich in hydroxycinnamic acids. The data obtained can be used to refine the indicators in the existing tables of the chemical composition of food products. Conclusion. The obtained data on the composition of fruit and berry products will make it possible to more correctly calculate the nutritional value of rations using questionnaire-survey methods. Combined analysis of the composition of biologically active substances in berries and fruits makes it possible to more reasonably make the choice of a particular product in the dietary correction of the ration of healthy and sick people.

[Alkaline transesterification CG-MS/MS method of monochlorpropanediol and glycidyl esters' determination in some edible fats, oils and fat blends on Russian market].

Monochlorpropanediol fatty acid esters (MCPDE) and glycidyl fatty acid esters (GE) are mainly considered to be processing contaminants and their concentration can rise during high temperature refining and deodorization of edible oils. Free forms formed during digestive hydrolysis of esters such as 3-monochloropropane-1,2-diol (3-MCPD), 2-MCPD and glycidol can provoke a negative effect on human health. Therefore the quantitative determination of MCPDE and GE in edible oils, fats and fat blends is needed. The aim - this manuscript deals with MCPDE and GE concentration measured as free 3-MCPD, 2-MCPD and glycidol in different edible oils, fats and fat blends of Russian market. Material and methods. 55 edible oil and fat samples sold on Russian market including refined and non-refined oils and fat blends such as spreads, dairy fat replacers, and margarines have been analyzed. Slow alkaline transesterification method with GC-MS/MS was used. Results. According to the data obtained, the highest concentrations of the contaminants were detected in fat blends: <0.10-5.03 mg/kg for 3-MCPD, <0.10-2.50 mg/kg for 2-MCPD and 0.1 5-11.17 mg/kg for glycidol. In palm oils and its fractions concentration of 3-MCPD was <0.10-6.61 mg/kg, 2-MCPD - <0.10-2.69 mg/kg and glycidol - <0.10-6.29 mg/kg. The content of glycidol in sunflower oils fluctuated in the range <0.10-1.19 mg/kg, 3-MCPD was <0.10-2.47 mg/kg, and 2-MCPD <0.10-0.67 mg/kg. Non-refined edible oils and olive oils had no or little MCPDE or GE. Conclusion. In this work we indicate high importance of monitoring MCPDE and GE in edible oils and fats both as ready-to-eat products and as ingredients prior to the Russian market release. There is strong need in mitigation of these process contaminants during fat blends manufacturing.

[Secondary lipid oxidation products. Human health risks evaluation (Article 1)].

The first article of the series describes possible applications of both proton nuclear magnetic resonance spectroscopy (1Н NMR) and Fourier transform infrared spectroscopy (FTIR) in food lipid thermo-oxidation analysis. Thermo-oxidation process is a source of various oxidation products. Some of them are known to be toxic, such as oxidized α,ß-unsaturated aldehydes and epoxidized linoleic acid derivatives. Today we know that routine nonspecific methods in lipid oxidation analysis are not informative, may provide incorrect results and procedures are long and laborious. Therefore it might be useful to find more reliable, accurate and informative physic-chemical methods measuring food lipid oxidation status. This paper is devoted to the most widely used in lipid analysis spectroscopic methods such as 1Н NMR and FTIR. It has been shown that 1Н NMR and FTIR provide more information on the types, formation and degradation time of compounds formed than wet chemistry methods. 1Н NMR gives qualitative and quantitative information on degraded and newly formed compounds and FTIR is able to measure a lot of standard oxidation indices with high accuracy. Both of them allow us to trace any compounds' evolution in lipid matrices in real time. Mention is made of their advantages for routine laboratory analysis.

[Comparative characteristics of in vivo models of hyperlipidemia in Wistar rats and C57Bl/6 mice].

In vivo simulation of lipid disorders (hyperlipidemia, obesity, metabolic syndrome, atherosclerosis) is of considerable interest to search for genomic, transcriptomic and metabolomic markers that allow for differential diagnosis, prognosis and selection of personalized diet therapy in patients with such pathology. The aim of the study was the development and characterization of basic biochemical parameters of in vivo models of alimentary hyperlipidemia in outbred rats and inbred mice. The experiment was con­ducted on 48 growing female Wistar rats, and 48 growing female mice of line C57Black/6, which were divided into 12 groups of 8 animals per group. Within 63 days the rats and mice of first (control) group received a balanced semi synthetic diet (BD), the animals of the second groups - high-fat diet (HFD) with 30% of the total fat by weight of dry feed, third groups - BD and fructose solution (Fr) instead of water, the fourth groups -HFD + Fr, fifth groups - BD supplemented with 0.5% cholesterol (Cho) by weight of dry feed, sixth groups - BD with Cho and Fr. The amount and composition of diets consumed were corrected during the experiment for their closest approach in calories. After removal of animals from the experiment there were determined the mass of internal organs, HDL, LDL, total cholesterol, triglycerides, glucose in blood plasma, total lipids and their fatty acid composition in liver, ghrelin, GIP, GLP-1, glucagon, leptin, PAI-1, resistin levels in blood plasma. It was found that in both species the liver is the most sensitive to nutritional imbalance, nutrient exerting the greatest impact on this was Fr. In rats, as compared to mice, there was significantly more pronounced shifts in lipoprotein spectrum in response to nutritional imbalances, especially to the consumption of additional Cho, which was manifested in an increase of LDL, decrease of HDL and magnification of atherogenic index. In the liver of rats fed diets with Cho, marked steatosis developed manifested in a disproportionate increase in the lipid content and accompanied by changes in their fatty acid composition, especially in the ratio ω6 to ω3 PUFAs. Changing of hormones - regula­tors of carbohydrate metabolism (GLP, glucagon) and ghrelin was significantly greater in mice than in rats as a result of consumption of additional Fr. Effect had the opposite direction in two species of Cho and Fr combining on leptin levels. The significance is dis­cussed of the revealed interspecies differences in the light of the characteristics of lipid and glucose metabolism in these two lines of animals that are the most common models of alimentary-dependent diseases.

[Enrichment of the rats diet with docosahexaenoic acid and astaxanthin: physiological and biochemical efficiency].

To investigate the effect of enrichment of the rats diet with polyunsaturated fatty acids (PUFA) ω-3 (220 mg docosahexaenoic acid per 1 kg of animal body weight per day) and astaxanthin (5 mg/kg body weight) on serum corticosterone concentration, physical fatigue, anxiety of rats after exhausting the load. During 30 days the rats of the test group received the diet in which the usual fat component comprising sunflower oil and lard (1:1) was completely replaced by the mixture of oils (high oleic sunflower (89%), coconut (6%), and marine oil from microalgae Schizochytrium sp. (5%) with a high content of docosahexaenoic acid with the addition of astaxanthin). Ratio of ω-6 and ω-3 PUFA in the lipid component of the experimental diet was 5.2:1 (n=12) and 135:1 in the diet of rats in the control group (n=12). DHA enrichment of the diet resulted in a significant 10-fold increase of the DHA liver content and ω-6 PUFA reducing (in particular of linoleic acid in 2.7-fold). No significant differences have been identified between the groups in terms of anxiety, estimated on the elevated plus maze at the beginning and on 24th day of the experiment. Results of the exhausting load on a treadmill (25th day) showed a significant reduction in physical fatigue in rats of the experimental group compared with the control group of rats: the number of contacts with the electrical grid was 4.2±0.9 versus 19.7±4.4, fulltime shock was 0.9±0.2 versus 3.3±0.8 sec. Significantly lower serum corticosterone concentration took place in the subjected to exhausting exertion animals receiving lipid module (15.0±3.9 ng/ml) compared to control animals (31.0±5.4 ng/ml). Thus, modification of the lipid component of the diet by its enrichment with DHA and astaxanthin led to decrease of the rat fatigue during exercise training (test treadmill) and prevent from the serum corticosterone raise, that indicates animal stress adaptation ability.