Methodology for the identification of bioactive and marker peptides in the organs of cattle and pigs

The development of general conception methodology for the meat-based functional food compositions is especially relevant today due to the growing consumers’ interest and attention to their health. This category of these food-products is intended for personal  ized nutrition of various age groups in the population, taking into account fortification of the food with nutraceuticals and with functional and metabolically active ingredients obtained from animal and vegetable source. Therefore, it was necessary to develop a certain tool for reliable identification of free peptides from the offals (by-products like hearts and aorta from Sus scrofa and Bos taurus) and from the ready-to-consume meat food (canned food) based on the free peptides, which food is potentially targeted to help with some issues in the human body. The authors proposed the methodology for identification of peptides weighing less than 5 kDa. This methodology has a row of significant advantages, such as a short time of analysis (90 minutes) and the possibility to prepare a large number of samples simultaneously (n=16). Analysis of bioactive peptides (BAPs) was performed by liquid chro  matography combined with time-of-flight mass spectrometry (Agilent 6545XT AdvanceBio LC/Q-TOF). The marker peptides were detected by a triple quadrupole mass spectrometer (Agilent 6410 Triple Quadrupole LC/MS). All peptide sequences were defined with the help of mass spectrometric data processing databases like PepBank, BioPep, AHTPDB. In this work from 39 to 269 peculiar soluble peptides were found, with an extraction level of 0.17–0.23%. The main fraction consisted of short peptides less than 1000 Da (71.0–98.0%). In experimental samples of pork hearts and arteries 7 peculiar marker peptides were identified. FFESFGDL  SNADAVMGNPK peptide obtained from the β-hemoglobin protein is of a special interest, as this peptide showed the maximum intensity of a signal. Presumably, this peptide can serve as an indicator of the blood presence in the finished food product. So it can serve as an assessment tool of bleeding degree of meat raw. For pork aortas a specific peptide TVLGNFAAFVQK was isolated from serum albumin, which turned out to be stable during heat treatment. This is also important for assessment of meat food that are subjected to high thermal exposure.The development of general conception methodology for the meat-based functional food compositions is especially relevant today due to the growing consumers’ interest and attention to their health. This category of these food-products is intended for personal  ized nutrition of various age groups in the population, taking into account fortification of the food with nutraceuticals and with functional and metabolically active ingredients obtained from animal and vegetable source. Therefore, it was necessary to develop a certain tool for reliable identification of free peptides from the offals (by-products like hearts and aorta from Sus scrofa and Bos taurus) and from the ready-to-consume meat food (canned food) based on the free peptides, which food is potentially targeted to help with some issues in the human body. The authors proposed the methodology for identification of peptides weighing less than 5 kDa. This methodology has a row of significant advantages, such as a short time of analysis (90 minutes) and the possibility to prepare a large number of samples simultaneously (n=16). Analysis of bioactive peptides (BAPs) was performed by liquid chro  matography combined with time-of-flight mass spectrometry (Agilent 6545XT AdvanceBio LC/Q-TOF). The marker peptides were detected by a triple quadrupole mass spectrometer (Agilent 6410 Triple Quadrupole LC/MS). All peptide sequences were defined with the help of mass spectrometric data processing databases like PepBank, BioPep, AHTPDB. In this work from 39 to 269 peculiar soluble peptides were found, with an extraction level of 0.17–0.23%. The main fraction consisted of short peptides less than 1000 Da (71.0–98.0%). In experimental samples of pork hearts and arteries 7 peculiar marker peptides were identified. FFESFGDL  SNADAVMGNPK peptide obtained from the β-hemoglobin protein is of a special interest, as this peptide showed the maximum intensity of a signal. Presumably, this peptide can serve as an indicator of the blood presence in the finished food product. So it can serve as an assessment tool of bleeding degree of meat raw. For pork aortas a specific peptide TVLGNFAAFVQK was isolated from serum albumin, which turned out to be stable during heat treatment. This is also important for assessment of meat food that are subjected to high thermal exposure

Biochemical Transformations of Lipide and Carbohydrat-Protein Nano Complex in Liquid Foodstuff

Abstract The process of enzymatic hydrolysis of dairy whey at presence of a polyfermental preparation pancreatin was investigated. The process of hydrolytic transformations of dairy proteins, fats and carbohydrates in comparison of a parallel estimation of changes of the sizes of the lipid, carbohydrates-protein formations which are being in nano region of area was studied

Biologically active peptides of meat and meat product proteins: a review. Part 2. Functionality of meat bioactive peptides

Biologically active peptides (BAP) are regarded as the main products of protein hydrolysis. BAP activity depends on the amino acid sequence molecular weight and chain length, type and charge of an amino acid at the N-terminus and C-terminus, hydrophobic and hydrophilic properties, spatial structure. They positively influence many systems of the human body, including the blood circulatory, nervous, immune, gastrointestinal and other systems. The health-improving effect of bioactive peptides is formed due to their antioxidant, antihypertensive, antithrombotic, immunomodulatory, antimicrobial, anti-allergic, opioid, anti-inflammatory, hypocholesterolemic and anticancer properties. Angiotensin-I-converting enzyme (ACE) inhibitory peptides are most studied due to their effect on blood pressure regulation. Unlike synthetic preparations, biologically active peptides do not have side effects and, therefore, can be used as their alternative. There is a growing commercial interest in peptides generated from meat proteins is in the context of health saving functional foods. The paper describes prospects, pros and cons of using bioactive peptides as functional food ingredients and biologically active food additives

Products of chemical reactions that occur during high-temperature heat treatment of the meat products

Recently the actively active studies have begun devoted to the accumulation of «harmful» substances in food products, which are supposedly accumulated in the body of a person who often consumes these products. Meat, as a source of full-featured animal protein, is especially popular in this aspect. For the preparation of meat products various types of heat treatment are used, almost each of which will inevitably lead to the destruction of some of the chemical compounds originally present in the product, and the formation of completely new chemical compounds, which can often be harmful to the human body. During high-temperature heat treatment (mainly frying), some chemical reactions in meat products occur, which lead to the formation of heterocyclic aromatic amines (HAA) in it. Due to the great variety of raw meat and cooking recipes, during the heat treatment HAA’s of various classes are formed, each of them will be peculiar for the particular type of raw material or recipe components (with the exception of MeIQx and PhIP, which always form during frying). The more complete understanding of the HAA’s formation mechanism will help study the products of Maillard reactions and Strecker degradation. In this work we studied the formation of HAA’s as a result of the cyclization of creatine and the detaching of water (dehydration) from it during temperature exposure. The classification of the compounds formed as a result of these reactions is presented and the main classes of the HAA obtained in result are considered. The questions of the influence of various factors on amount of HAA formed, such as the fat content, the introduction of Fe2+, Fe3+, are raised. In the future it is necessary to conduct studies of the quantitative content of HAA in meat products to complement the already actively ongoing work on the study of xenobiotics consumed by humans with food, which will give a more comprehensive picture of the carcinogens content in food products

Effect of organically bound iodine in cattle feed on health indicators

Currently, the problem of iodine deficiency is actual in the world, which may cause a large number of diseases and disorders. The problem of iodine deficiency for humans may be partly solved by enriching agricultural products with iodine, i.e. by providing animals with an increased intake of iodine during their growth. Theoretically, the most effective way to use iodine is the form bound to tyrosine, since diiodotyrosine has been proven to be a thyroxine precursor. Taking it into account, a supplement was developed containing iodine organically bound to tyrosine and histidine. In this work, we studied the effect of this supplement introduced into the diets of cattle on biochemical parameters of animal blood. In the test group, which received the supplement with organically bound iodine, an improvement in nitrogen metabolism was noted compared to the control group. This was most clearly demonstrated by the content of urea, since in the test group, its content decreased by ≈15 percentage points, and by the content of creatinine, since its increase in the test group was more than 20 percentage points. Differences in the parameters of carbohydrate and lipid metabolism between treatments were also noted, as in the blood of animals from the test group, the content of cholesterol, triglycerides, phospholipids, glucose and malondialdehyde was lower than in the control group. In mineral metabolism and morphological parameters, there was no significant difference between treatments. Among the indicators of pigment and hormone metabolism, it is important to note the reduced content of cortisol in the blood of animals from the test group. Its level was lower by ≈17.23 percentage points compared to the control group. A decrease in cortisol levels indicated a lower stress load in the test group. In general, studies have shown that the use of a feed supplement containing organically bound iodine has a positive effect on the metabolism of animals.Currently, the problem of iodine deficiency is actual in the world, which may cause a large number of diseases and disorders. The problem of iodine deficiency for humans may be partly solved by enriching agricultural products with iodine, i.e. by providing animals with an increased intake of iodine during their growth. Theoretically, the most effective way to use iodine is the form bound to tyrosine, since diiodotyrosine has been proven to be a thyroxine precursor. Taking it into account, a supplement was developed containing iodine organically bound to tyrosine and histidine. In this work, we studied the effect of this supplement introduced into the diets of cattle on biochemical parameters of animal blood. In the test group, which received the supplement with organically bound iodine, an improvement in nitrogen metabolism was noted compared to the control group. This was most clearly demonstrated by the content of urea, since in the test group, its content decreased by ≈15 percentage points, and by the content of creatinine, since its increase in the test group was more than 20 percentage points. Differences in the parameters of carbohydrate and lipid metabolism between treatments were also noted, as in the blood of animals from the test group, the content of cholesterol, triglycerides, phospholipids, glucose and malondialdehyde was lower than in the control group. In mineral metabolism and morphological parameters, there was no significant difference between treatments. Among the indicators of pigment and hormone metabolism, it is important to note the reduced content of cortisol in the blood of animals from the test group. Its level was lower by ≈17.23 percentage points compared to the control group. A decrease in cortisol levels indicated a lower stress load in the test group. In general, studies have shown that the use of a feed supplement containing organically bound iodine has a positive effect on the metabolism of animals

Molecular genetic methods for identifying raw materials in meat products: Diversity, opportunities and prospects

In the current economic situation, after easing the Covid pandemic restrictions, almost all laboratories, which are focused on evaluation of the conformity of food products, have faced issues in supplying for their laboratories. In this regard, in the last years many laboratories have been forced to validate new approaches and introduce new methods for assessing conformity of the food products. Very often it is not possible to use only one method to resolve the issue of the food product ingredients, especially for the purpose of traceability of their names and the used raw materials, listed on the label. Survey of the raw food materials to determine whether they correspond to the type name is a simpler task, in contrast to survey of the multicomponent food product. Many researchers have to estimate the opportunities and feasibility of application of various methodologies in their workplaces. Therefore, this review is relevant for the researchers in this field, as it focuses on aspects and special features of similar methodologies. The prospect of molecular genetic methods for identification of the raw materials used for manufacturing of meat products is presented below. This review also represents characteristics of methods for identification of the sources of raw materials used for the manufacturing of the meat products, based on the recognition of species-specific sections within the nucleic acids structures. The variety of methods (hybridization methods, polymerase chain reaction, different types of isothermal amplifications, methods using CRISPR/Cas systems), the principles of their implementation, and achieved analytical characteristics are considered. The capacities and competitive potential of various methods are discussed, as well as approaches being developed to overcome the existing limitations.In the current economic situation, after easing the Covid pandemic restrictions, almost all laboratories, which are focused on evaluation of the conformity of food products, have faced issues in supplying for their laboratories. In this regard, in the last years many laboratories have been forced to validate new approaches and introduce new methods for assessing conformity of the food products. Very often it is not possible to use only one method to resolve the issue of the food product ingredients, especially for the purpose of traceability of their names and the used raw materials, listed on the label. Survey of the raw food materials to determine whether they correspond to the type name is a simpler task, in contrast to survey of the multicomponent food product. Many researchers have to estimate the opportunities and feasibility of application of various methodologies in their workplaces. Therefore, this review is relevant for the researchers in this field, as it focuses on aspects and special features of similar methodologies. The prospect of molecular genetic methods for identification of the raw materials used for manufacturing of meat products is presented below. This review also represents characteristics of methods for identification of the sources of raw materials used for the manufacturing of the meat products, based on the recognition of species-specific sections within the nucleic acids structures. The variety of methods (hybridization methods, polymerase chain reaction, different types of isothermal amplifications, methods using CRISPR/Cas systems), the principles of their implementation, and achieved analytical characteristics are considered. The capacities and competitive potential of various methods are discussed, as well as approaches being developed to overcome the existing limitations

The formation of flavoring characteristics of meat products by changing the chemical composition of food compositions

The article presents the results of the study of changes in flavour characteristics when using corrective additives. Monosodium glutamate, ribotide, yeast extract and hydrolysate of vegetable soy protein were used as flavoring additives (FA). To assess the effect of composition of meat product recipe, as well as the method of FA‑introduction on taste intensity, the recipes of model meat systems with partial replacement of meat raw materials were used. Pork fat, soy protein and potato starch were used as meat substitutes. The effect of recipe composition on the content of non-volatile substances of aroma was accessed. It is shown that replacement of pork by pork fat in the recipe by 20–40% led to a sharp decrease in the concentration of aromatic substances and a decrease in intensity of taste of the finished product several times. The ways for taste correction using FA was studied. For this, a chopped semi-finished product — minced meat was prepared from chilled whole-muscle pork and 0.05% of each FA was added. It is shown that the dynamics of changes in the content of free amino acids is the most pronounced when using monosodium glutamate not as a mono-additive, but in compositions: monosodium glutamate with yeast extract and monosodium glutamate with ribotide. A pool of chemical compounds involved in the formation of taste and aroma of products was detected. The main components were derivatives of C6–C24 fatty acids, as well as a significant number of other biochemical compounds, mainly substituted amines, amides, alcohols and ketones, with a content ranging from 0.001 to 0.2 mg/kg. The results of organoleptic analysis showed that the most delicious and attractive samples were those containing monosodium glutamate with yeast extract and monosodium glutamate with ribotide

A procedure for the change point problem in parametric models based on phi-divergence test-statistics

This paper studies the change point problem for a general parametric, univariate or multivariate family of distributions. An information theoretic procedure is developed which is based on general divergence measures for testing the hypothesis of the existence of a change. For comparing the accuracy of the new test-statistic a simulation study is performed for the special case of a univariate discrete model. Finally, the procedure proposed in this paper is illustrated through a classical change-point example

БИОИНФОРМАТИКА — ИНСТРУМЕНТ ИНТЕРПРЕТАЦИИ ПРОТЕОМНЫХ ПРОФИЛЕЙ БЕЛКОВ МЯСА

Proteomic technologies have proven very effective for detection in meat products of biochemical changes, such as changes in heat resistant and species-specific proteins that could be relevant bio-markers.In the work presented in this report (for the period of 2013–2016), several tissue-specific proteins were detected in the samples of meat and specially developed meat products using proteomic technologies and identified as individual biomarkers in meat product control.The existence of a large number of different proteins resulted in the need to create information arrays — databases (or banks). Currently, there are a number of general and specialized databases that are available online to anyone interested. When studying protein proteomic profiles, many scientists stop at the stage of two-dimensional electrophoregrams sometimes even without ideas about the future prospects of using modern instruments and bioinformation resources to confirm or refute their hypotheses, and sometimes just to identify. This overview shows the chain of actions that allows going from profiling proteins in the gel to a specific interpretation of the results. Studies in this field have enabled formulating and significantly expanding the approaches to the identification and quantification of protein markers of quality, functionality and safety of meat raw material (detection of falsification) in the finished meat products. Based on the obtained data, the information was systematized using bioinformatics techniques with creation of the unique Atlas «Proteomic profiles of farm animal meat proteins.»Протеомные технологии оказались весьма эффективными для выявления в  мясных продуктах биохимических изменений, таких как изменения термоустойчивых и видоспецифичных белков, способных стать соответствующими биомаркерами. В работе, представленной в  данном обзоре (в период с 2013–2016 гг), с помощью протеомных технологий в исследуемых образцах мяса и в специально выработанных мясных изделиях, было определено несколько тканеспецифичных белков, которые были определены как индивидуальные биомаркеры при контроле мясных изделий. Существование огромного количества разнообразных белков привело к необходимости создания информационных массивов — баз (или банков) данных. В настоящее время существует множество общих и специализированных баз данных, которые доступны в Интернете каждому желающему. При исследовании протеомных профилей белков, многие ученые останавливаются на этапе получения двумерных электро-фореграмм, не имея порой даже представления о дальнейших перспективах использования современных инструментальных и биоинформационных ресурсов, позволяющих подтвердить или опровергнуть их гипотезы, а порой просто идентифицировать. В данной статье представлена цепочка действий, позволяющих пройти путь от получения профиля белков на геле, до конкретной интерпретации полученного результата. Выполнение исследований в данном направлении позволило сформулировать и  значительно расширить подходы к идентификации и количественному определению белковых маркеров качества, функциональности и безопасности мясного сырья (выявления фальсификации) в готовых мясных продуктах. По полученным данным систематизирована информация с помощью методов биоинформатики, позволившая создать уникальный Атлас «Протеомные профили белков мяса сельскохозяйственных животных»