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Antioxidants play a very important role in the food industry. Recently, both science and industry have shown substantial preference for natural antioxidants, including searching for antioxidant substances from natural sources without undesirable side effects. The purpose of this study was to evaluate the effect of adding Allium cepa husk extract at a volume of 68 or 34 μL/g of unsalted blanched materials to replace 34% and 17% of the beef broth, respectively, which corresponded to a total antioxidant capacity (TAC) of 44.4 or 22.2 μmol-equiv. Q/100 g meat pté (i.e., 13.42 or 6.71 mg of quercetin/100 g meat pté), on the quality and safety indicators of the developed meat pté. The TAC according to a ferric reducing antioxidant power assay, thiobarbituric acid reactive substances, and physicochemical and microbiological characteristics were determined during the storage of the meat pté. Proximal and UPLC-ESI-Q-TOF-MS analyses were also performed. The addition of yellow onion husk ethanolic extract to the meat pté at both volumes allowed the maintenance of an increased content of antioxidants, which contributed to a decrease in the generation of secondary products of lipid peroxidation for 14 days of storage at 4 ∘C. The results of the microbiological analyses showed that the developed meat ptés were safe according to all indicators of microbial spoilage within 10 days of production. The results supported the use of yellow onion husk extract in the food industry to contribute to improving the functionality of meat products, developing products for a healthy lifestyle, and providing clean-label foods without or with a minimal content of synthetic additives.

Because of the increased demand for processed meat, there is an urgent need to introduce specific identification methods. Strategies such as molecular genetics and the physical condition of meat are used to quickly explore multi-component products. However, a single methodology does not always unambiguously classify a product as counterfeit. In laboratory practice, as a rule, screening techniques are rarely used in the first stage, followed by arbitration. This work aimed to study individual methodologies using artificially falsified meat samples as examples and to identify their composition based on muscle tissue. For the experiments, the three most common types of raw meat were selected: pork, beef, and chicken. The calculation of the content of muscle tissue was carried out according to the BEFFE method. The study of muscle protein was carried out by ICA, ELISA, PCR, microstructural analysis, and mass spectrometric identification. In this connection, we proposed a multilevel control system for multicomponent meat products. Both classical methodologies, such as calculation by prescription bookmarks (BEFFE) and microstructural analysis, and approaches of highly sensitive methodologies, such as identification of muscle tissue by marker peptides (LC/MS-MRM) and semi-quantitative PCR analysis, were evaluated.

Proteases are produced by a wide variety of microorganisms and have a wide range of applications. Peptidase M9 from Aeromonas salmonicida is a 915 amino acid residue protein. Earlier, we cloned the 864 bp C end of the gene. (col2 gene) encoding the actual peptidase domain of 238 aa in the vector plasmid pPic9K. It was demonstrated that the recombinant Pichia pastoris strain, created on the basis of this plasmid, has a specific enzymatic activity against collagen. The aim of this work was to further optimize the expression of the col2 gene of the M9 peptidase domain from Aeromonas salmonicida in Pichia pastoris, isolation, purification, and characterization of the protein.First, the col2 gene was optimized based on the use of P. pastoris codons. Secondly, a study of collagenase activity with 4 different signaling sequences was carried out. It was shown that the strain for which the collagenase gene had the P. pastoris alpha peptide as a signal sequence was shown to have the maximum activity. One of the approaches leading to an increase in the yield of the target protein is the cloning of the gene encoding the chaperone proteins. We have created strains containing, in addition to the collagenase gene, also the pdi and hac chaperone genes. It was shown that the collagenase activity in the strain containing the pdi chaperone gene was 2–3 times higher than in the strain containing only the col2 gene. So, the Pichia pastoris strain was created - the producer of the recombinant peptidase M9 from Aeromonas salmonicida, the target protein was isolated and characterized.Research conducted as part of the state assignment of Gorbatov Federal Research Center for Food Systems of Russian Academy of Sciences 0437-2019-0001.

Unfair production and products that do not comply with the declared labeling are currently an acute problem in the field of technical regulation, including with regard to food safety and quality. Given the high added value and multicomponent composition, finished meat products are among the most susceptible to adulteration. Despite the best efforts of regulatory agencies to counteract these inconsistencies, the hidden substitution of cheaper or lower-grade meats is still widespread. One of the main tasks facing research laboratories and testing centers today is the detection of falsification of food products, as well as standardization and certification of techniques necessary to solve such problems. The manufacturer, aware of the current control methods, can go to the deception, using vegetable protein, new unregistered feed additives. To determine the complex changes that occur in products, it is necessary to use methodological approaches in which it is possible to reliably determine these changes. The paper presents an overview of the most commonly used methodologies for assessing the component composition of meat products. Quality assessment of meat products includes control of components of finished products. The most difficult task is to determine the proportion of muscle protein in multicomponent meat products that have undergone heat treatment.

Insufficient quality and replacement of raw meat for vegetable are quite urgent problems. This is due to the expansion of rights and the emergence of new meat processing enterprises. The manufacturer may use vegetable protein, new unregistered feed additives. This paper presents the LC-MS method for detecting and quantifying two different types of meat (beef and pork) in a biological matrix of structureless minced meat. After isolating the proteins and digesting them with trypsin, species-specific peptide markers were selected for each animal species for quantitative determination. High-performance liquid chromatography (HPLC) Agilent 1260 with triple quadrupole mass spectrometer detector 6410 was provided for ease of usage and improve veracity of the analysis. This analysis used three stable peptides for the quantitative determination of two putative muscle biomarkers for pork and beef in minced meat with a 25-minute analysis. Good sensitivity was established (LOD 0.23–0.29%) with the ability to quantify the muscle tissue of each animal species (LOQ 0.70–0.86%) using special calibration curves.

Functional foods promote health benefits in human metabolism, with bioactive compounds acting as therapeutic agents. The aim was to investigate the biological effects of a pâté made of pork hearts and aortas, minced, sterilised and packed in tins. Adults (61–66 years old) with a body mass index of 26.4–60.7 kg/m2 (n = 36) were randomly divided into two groups: one group consumed a low-calorie diet (LCD), while the other consumed an LCD with the developed meat product (MP) for 28–30 days. Serum biochemical parameters, anthropometry and blood pressure were measured. Consumption of an LCD + MP by experimental group participants helped to maintain reduced cholesterol levels. The difference in total cholesterol was significantly different (p = 0.018) from that of the control group, mainly due to the difference in low-density lipoprotein cholesterol (p = 0.005). Six peptides with potential cholesterol-binding properties and four peptides with potential antioxidant activity were identified in the MP, while elevation of the content of two peptides with potential angiotensin-converting enzyme-inhibitory activity was detected in patients’ plasma. Intervention with the MP can be considered as a supportive therapy to the main treatment for medical cardiovascular diseases due to a positive effect on serum cholesterol.

Functional, particularly personalized meat-based foods are of more in demand by a consumer today. Functional additives, such as plant components and animal proteins from bovine or porcine tissues have been successfully used. With many ingredients added to foods, it is important to provide quality and composition monitoring to confirm the products’ authenticity, to identify undeclared or rarely used types of raw meat in product formulations. For example, if animal heart tissue is a component of a product formulation or if aorta tissue presents in a product due to improper trimming. Different methods are used to identify raw materials, including new approaches in proteomics and peptidomics that are considered the most effective modern methods nowadays. The purpose of the study is meat product composition analysis and special biomarker peptide identification to confirm the presence of heart and aorta tissue in a finished meat product. Over 20 amino acid sequences were checked based on earlier obtained data. Those amino acid sequences were analyzed with a high-performance liquid chromatography with mass spectrometric detection as described. The MS settings were selected using the Skyline. Signal-to-Noise ratio (S/N) over 10 units were used to choose the best peptide candidates. Seven peptides were found in porcine hearts. The best candidate was peptide VNVDEVGGEALGR (S/N - 73.10±5.3) from β-Hemoglobin. Two marker peptides from serum albumin were selected for pork aorta: TVLGNFAAFVQK (S/N 53.51±2.4) and EVTEFAK (S/N 31.69±4.1). These biomarkers showed the best detection and specificity. The multiply reaction monitoring method made it possible to identify the most/best specific peptides—biomarkers that could confirm the heart and/or aorta in meat products. The method can be used for comparative research or identification of best peptides that are specific to any type of animal tissue. The work was supported by the Russian Science Foundation, project no. 16-16- 10073.

One of the ways to address the food quality issues facing the industry is the development of standardized and certified methods related to the conduct of in-depth studies of biochemical indicators of quality and safety of meat and meat products. The world laboratory practice in the field of food quality and safety shows a constant expansion of the list of controlled indicators of food raw materials and products. An important feature of the modern period in the development of biomedical and biotechnological research is the introduction of a whole complex of postgenomic technologies, which are based on a systematic approach to the study of the functioning of the mammalian proteome in various physiological and pathological conditions, including the formation and development of alimentary-dependent pathologies. In this regard, the problem of multilateral study of food products, in particular their identification, is the most relevant, because the modern technology of their production has undergone significant changes and requires the development of “gentle “ processing modes. They concern raw materials and auxiliary materials used at all stages of production. This and new technologies of production of protein products from plant raw materials, as well as the introduction of food raw materials and food additives of artificial origin and the excess introduction of additives of plant and animal origin can cause falsification of products, as well as affect the health of the consumer. Food quality assessment includes the control of components of finished products. It is most difficult to determine the proportion of muscle protein in multi-component meat products that have undergone heat treatment. Therefore, in practice, when assessing the quality of food products, there is a need to identify its real composition in accordance with the declared normative documents. Currently, a promising area of research in the field of determining the composition of finished food is the selection of biomarkers of various components. Therefore, it is important to develop a methodology for the identification of biochemical changes in food raw materials under the influence of technological factors using modern research methods. This paper provides an overview of the protein and peptide analysis methodology, including the latest technologies that are becoming increasingly important.