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Developing the technology and evaluating the biological value of the peptone from secondary products of processing of fish raw material of the arctic region

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The results of microbiological and toxicological tests have proved the safety of the bone-muscular cod waste (BMW). Waste (humerus with meat cuts) from cod cutting on fillets contains 18,95 % of a full-grade animal protein and insignificant amount of fat (0.15 %).

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Nội dung Text: Developing the technology and evaluating the biological value of the peptone from secondary products of processing of fish raw material of the arctic region

  1. International Journal of Mechanical Engineering and Technology (IJMET) Volume 10, Issue 03, March 2019, pp. 1880-1893. Article ID: IJMET_10_03_191 Available online at http://www.iaeme.com/ijmet/issues.asp?JType=IJMET&VType=10&IType=3 ISSN Print: 0976-6340 and ISSN Online: 0976-6359 © IAEME Publication Scopus Indexed DEVELOPING THE TECHNOLOGY AND EVALUATING THE BIOLOGICAL VALUE OF THE PEPTONE FROM SECONDARY PRODUCTS OF PROCESSING OF FISH RAW MATERIAL OF THE ARCTIC REGION Ludmila Kazimirovna Kuranova, Vlasimir Aleksandrovich Grokhovsky, Yulia Viacheslavovna Zhivlyantseva and Vasily Igorevich Volchenko Department of Food Production Technology Institute of Natural Science and Technology Federal State Educational Institution of Higher Education "Murmansk State Technical University" Murmansk, Russian Federation ABSTRACT The results of microbiological and toxicological tests have proved the safety of the bone-muscular cod waste (BMW). Waste (humerus with meat cuts) from cod cutting on fillets contains 18,95 % of a full-grade animal protein and insignificant amount of fat (0.15 %). Thus, the possibility of using them as a protein raw material for producing peptones has been proved. The technology of obtaining peptone from the secondary fish raw material - bone-muscular waste from the cutting of cod fish - has been developed and optimized. Using the cryoextrusion method in the technology of peptone production at the stage of grinding waste is proposed. Using of the enzyme (protosubtilin G3X) in hydrolysis is substantiated; proteolytic activity is established, it is 560,77 μmol TYR / g, the optimal temperature of enzymatic hydrolysis of BMW is (45±1) ˚С. Using the theory of experiment planning and computer modeling, a series of works was carried out to optimize the stage of enzymatic hydrolysis of fish wastes. Nearly optimal hydrolysis parameters were found: enzyme concentration of 1,33 % to total waste weight, duration of hydrolysis process of 3 hours. The chemical and biochemical quality characteristics of the enzymatic peptone obtained by the optimized technology have been researched. It was found that the mass fraction of protein in the product is of 92,27 %, water is of 4,7 %, sodium chloride is of 2,6 %, fat is of 0,3 %. The amino acid composition has been determined with the method of high- performance liquid chromatography. It made it possible to calculate the biological value of peptone. Tryptophan has been established to be the only limiting amino acid http://www.iaeme.com/IJMET/index.asp 1880 editor@iaeme.com
  2. Ludmila Kazimirovna Kuranova, Vlasimir Aleksandrovich Grokhovsky, Yulia Viacheslavovna Zhivlyantseva and Vasily Igorevich Volchenko in the peptone protein (score is 70 %); which characterizes the peptone as a sufficiently balanced protein product, which can be recommended for use in food as a complete protein additives to food. Keywords: Bone-muscular cod waste, Protosubtilin, Enzymatic peptone, Amino acid composition, Triptophan, Balanced protein product. Cite this Article Ludmila Kazimirovna Kuranova, Vlasimir Aleksandrovich Grokhovsky, Yulia Viacheslavovna Zhivlyantseva and Vasily Igorevich Volchenko, Developing the Technology and Evaluating the Biological Value of the Peptone From Secondary Products of Processing of Fish Raw Material of the Arctic Region, International Journal of Mechanical Engineering and Technology, 10(3), 2019, pp. 1880-1893. http://www.iaeme.com/IJMET/issues.asp?JType=IJMET&VType=10&IType=3 1. INTRODUCTION The problem of providing the population of Russia with the foodstuffs with the increased nutrition and biological value, balanced іn micronutrients and containing biologically active substances positively influencing functions of organs and tissues of the person, is faced now by national economy of Russia and it is acute and very urgent [1, 2]. The need for protein is an evolutionary dominant in human nutrition, which is due to the need to provide the optimal physiological level of essential amino acids. The quality of the protein – its biological value – is determined by the presence in it of a complete set of essential amino acids in a certain ratio. Essential amino acids are represented by amino acids with carbon branch chain – leucine, isoleucine and valine, aromatic – phenylalanine, tryptophan and aliphatic – threonine, lysine and methionine. Since the body synthesizes cysteine and tyrosine from methionine and phenylalanine, respectively, then the presence in food in sufficient quantities of these two amino acids reduces the need for essential precursors. Partially nonessential amino acids include arginine and histidine, as they are synthesized rather slowly in the body. A young and growing organism is particularly in need of histidine. The absence of at least one essential amino acid in food causes a negative nitrogen balance, disruption of the central nervous system, growth stop and severe clinical consequences such as vitamin deficiency. Lack of one essential amino acid leads to incomplete assimilation of others [3]. There is a need to develop new technologies for the development of non-traditional nutrition for the fishing industry — protein products - peptones, which can be successfully used as a protein component of foodstuff, feed of farm animals, birds, and aquaculture facilities, nutritional and diagnostic microbiological environments, pharmaceutical and cosmetic products [4, 5]. Peptons are large protein fragments that are formed during hydrolysis. It is a protein-containing product, the amount of "total" protein in which is not less than 85 %, and "true" protein – not less than 75 %, i. e. protein isolate [3]. The creation of new types of products based on peptones will partly solve not only the problem of environmental pollution, but also the problem of protein deficiency. It is known that enzymatic hydrolysis of fish proteins forms a mixture of free amino acids, di -, tri-and oligopeptides, the number of polar groups and the solubility of peptone increases, and therefore changes the functional characteristics of proteins, improving their functional properties and biological value. This is important if the peptone is used as a food ingredient [6]. http://www.iaeme.com/IJMET/index.asp 1881 editor@iaeme.com
  3. Developing the Technology and Evaluating the Biological Value of the Peptone From Secondary Products of Processing of Fish Raw Material of the Arctic Region The pace of modern life dictate their own conditions and, unfortunately, people do not always manage to eat the right, balanced food, observe the regime, take sports. In nature, there are only a few equivalent sources of high-grade protein of the highest biological value: milk, egg, meat, fish and soy. The finiteness of natural resources leads to the fact that the increase in the volume of animal protein produced lags behind the growing needs of the population. Today, the volume of fish catch in Russia in absolute terms is 4.2 million tons. However, up to 28 % of the catch (or more than 1 million tons) is not used for food purposes. Due to the development of new industrial projects in the Arctic Region, a significant increase in traffic volume along the Northern Sea Route can be currently observed. To protect the environment against polluting emissions from shipping, now, requirements to the quality of marine fuel are being updated and tightened, new environmental regulations worked out. The strictest regulations are applied to some air pollutant Emission Control Areas (ECA). In Europe, ECA include Baltic Sea and Northern Sea areas, in the Northern America – Pacific and Atlantic coast areas [7]. One of the most significant examples of intensive anthropogenic load on the natural landscape is the metallurgical complex “Severonikel” (Murmansk Region). The metallurgical complex is located on the Kola Peninsula in close proximity to the administrative border of the Russian Federation with Finland [31]. Deposition of heavy metals with the atmospheric precipitation in the form of acid rain is the main way of their ingress from atmospheric air into soil and surface water bodies [8]. In this regard, of particular importance and relevance there are the studies aimed at the development of safe food protein products from hydrobionts, involving in the processing of secondary products formed in the processes of traditional processing of hydrobionts, in particular, bone-muscular waste from cutting cod fish species on fillets. Atlantic cod (Gadus morhua) is a traditional object of fishing in the Arctic region. The chemical composition of cod is well studied. The average protein content in the muscle tissue of cod varies within 16– 18% [9, 10]. The component composition of tissues is characterized by low fat content (less than 1 %). Waste from cutting into fillets include: skin cover and scales, fins, rib bones, vertebral bone, head, entrails and tail. Fish waste is an important reserve of food raw materials, which is often underestimated [11, 12]. Currently, in the literature there are works devoted to the study of their chemical and biochemical composition [4, 13, 14]. However, in- depth study of the properties and possibilities of deep processing of bone-muscular waste from cutting of traditional raw materials was practically not carried out. This implies the purpose of this work, which is associated with the study of bone- muscular waste from cutting cod for its subsequent use in the creation of innovative technologies for processing hydrobionts — development of peptone technology, optimization of process parameters, the study of the quality of the fish peptone, the study of its biological value and directions of use. The possibility of using peptones obtained from secondary fish raw materials will allow to preserve valuable protein products in the food ration, reduce the volume of non-recyclable waste and expand the range of products from hydrobionts used for food, feed and medical purposes. 2. OBJECTS, MATERIALS AND METHODS OF RESEARCH The objects of research were: bone-muscular cod waste (BMW); obtained by cutting fish into a carcass by separating the head with the nape (cod caught by the Public Joint-Stock Company "Murmansky Trailing Fleet" in the fishing areas of the Barents sea, was frozen and delivered to the port of Murmansk, where it was stored for 1 month at a temperature not http://www.iaeme.com/IJMET/index.asp 1882 editor@iaeme.com
  4. Ludmila Kazimirovna Kuranova, Vlasimir Aleksandrovich Grokhovsky, Yulia Viacheslavovna Zhivlyantseva and Vasily Igorevich Volchenko higher than minus 18°C); protosubtilin G3X – industrial enzyme preparation – a product of bacteria strain Bacillus subtilis; pepton (PBMW), obtained in the process of enzymatic hydrolysis of BMW according to the technology developed by the authors [15]. Chemical reagents, amino acid standards for chromatography and standard samples for atomic absorption spectroscopy were purchased from Sigma Aldrich (Germany) for chemical analysis [30]. Chemical and biochemical methods accepted in scientific researches are used in the work. The mass fraction of water, lipids, protein, amine nitrogen, minerals of raw materials was determined by standardized methods. The protein content was determined by the Kjeldahl method using equipment consisting of two elements: BLOCK–DIGEST–12 for sample mineralization and automatic distillation unit PRO–NITRO A (J.P. Selekta, Spain). The fat content was determined by the Soxhlet method using the Selecta DET/GRAS extractor (Spain). Amino acid composition of the peptone was determined by hydrolysis of the sample with hydrochloric acid or alkali upon heating [16], subsequent modification of the resulting amino acids with phenylisothiocyanate, separation of phenylthiocarbamyl amino acid derivatives on a column with reversed phase followed by registration with SPD-20AV spectrophotometric detector on liquid chromatograph LC-20 Prominence of Shimadzu (Japan) [32]. The content of lead, cadmium, arsenic, mercury was determined by atomic absorption spectrometry [17] on the atomic absorption spectrometer Shimadzu AA-6300 (Japan). Proteolytic activity of the enzyme preparation was determined by the method based on spectrophotometric determination of dissolved amino acids and peptides formed during enzymatic hydrolysis of casein [18]. The degree of hydrolysis was determined by calculation as the ratio of the mass fraction of amine nitrogen to the mass fraction of total nitrogen in the hydrolyzate [19]. Amino acid score (AAS) [19, 21] was calculated by the formula: 𝑚1 AAS= 𝑚2 ∙ 100 , % (1) where: m1 – content of essential amino acid in 1 g of peptone protein, mg/g of protein, m2 – content of essential amino acid in 1 g of reference protein, mg/g of reference protein. When microbiological control of the samples we tested (according to standard microbiological techniques) for the mesophyll aerobic and optional-anaerobic microorganisms microorganisms (QMAFAnM), the presence of coliform bacteria (coliforms) of the genus Staphylococcus aureus, the presence of pathogenic organisms, including of the genus Salmonella, Listeria monocytogenes, yeast and mold fungi. In determining the optimal parameters of enzymatic hydrolysis, a second-order rotatable composite plan was used for two factors [22]. Mathematical data processing was performed using DataFit version 9.1 2.1. Statistical analysis The experiments were repeated three times, and the data obtained were subjected to single- factor analysis of variance (ANOVA) using Origin Pro 8.0. Differences between averages were considered significant at p  0,05. 3. RESEARCH RESULTS AND DISCUSSION http://www.iaeme.com/IJMET/index.asp 1883 editor@iaeme.com
  5. Developing the Technology and Evaluating the Biological Value of the Peptone From Secondary Products of Processing of Fish Raw Material of the Arctic Region Currently, there are many technologies for obtaining peptones. As a raw material, any natural proteins full of amino acid composition are used, the sources of which are blood and its components; tissues of animals and plants; waste of dairy and food industries; food and low- value food products obtained in the processing of various species of animals, birds, etc. [23]. At the same time, there is another promising source that can be used for the production of peptones – bone-muscular waste generated during the cutting of cod fish species in the sea. According to the existing standards of fishing waste should be frozen and delivered to the shore for disposal, but due to the closure of plants for the production of fish meal, the issue of its processing has not yet been resolved. The authors conducted a comprehensive study of the quality of waste obtained from cutting cod. Analysis of the chemical composition found that the protein content in the waste is 20.2 % (due to a significant number of cuts of meat), fat content - 0.31 %. The content of toxic elements in the bone-muscular cod waste was: lead - less than 0.05 mg/kg, cadmium - less than 0.005 mg/kg, arsenic - less than 0.05 mg/kg, mercury - 0.028 mg/kg, copper - 0.83 mg / kg. Microbiological studies have established the absence of dangerous microorganisms for the human body: coliform bacteria, Staphylococcus aureus, pathogenic Salmonella and Listeria. The results of chemical and microbiological tests indicate the safety of bone-muscular cod waste, and thus confirm the possibility of using them as additional raw materials in the production of food. Taking into account the results of their own research and literature data, the authors developed a technology for the production of peptones from bone-muscular cod waste by enzymatic hydrolysis. The basic technological scheme of production of peptone from bone-muscular waste is reduced to the following operations: thawing of bone-muscular waste, crushing, fermentation, purification from the bone residue and non-hydrolyzed protein by acidification and alkalization with gradual filtration, drying. One of the stages of the technological process is the production of ground minced mass. To this end, traditionally the raw material is subjected to defrosting and grinding. This is quite time-consuming and lengthy process, during which there is a decrease in protein content due to the loss of cellular juice. To reduce energy and labor costs, the method of grinding raw materials by cryoextrusion on the layout of the plunger with a die, 50 mm in diameter, cooled to the temperature of the raw material was used. The method of cryoextrusion is a crushing of frozen raw materials by punching through a cooled die hole, and cutting the fibers of the muscle tissue of the raw materials by the ice crystals [24]. The use of cryo-grinding eliminates the defrosting of raw materials, which avoids the loss of raw materials and preserve its properties. Studying the manufacturing process of enzymatic peptone, it is impossible to take into account all the conditions affecting the fermentation process. Important factors affecting the hydrolysis process are the temperature and duration of the process, as well as the quality and quantity of the enzyme administered. For protein hydrolysis as an enzyme preparation, various enzymes can be used. So Zubtsov V. A and others [25] carried out pepsin and trypsin hydrolysis of animal products. Gastric mucosa and pancreas were used as the enzyme source. Sultanov and others [26] conducted pepsin hydrolysis, as raw materials they uses fish (sprat, pollock, salmon) or feeding meal. E. Dimova and others [27] carried out the hydrolysis of the skin and blood of a calf by using alkaline proteases of bacterial origin (Bacillus subtilis strain DY). Antipova L. V. and others [28] conducted hydrolysis of secondary products of cutting pond fish using the http://www.iaeme.com/IJMET/index.asp 1884 editor@iaeme.com
  6. Ludmila Kazimirovna Kuranova, Vlasimir Aleksandrovich Grokhovsky, Yulia Viacheslavovna Zhivlyantseva and Vasily Igorevich Volchenko enzyme protosubtilin G3X and collagenase. Novikov V. Yu. and others [29] hydrolyzed the tissue of Atlantic cod using an enzyme preparation derived from the hepatopancreas of the Alaska king crab Paralithodes camtschatica. Due to the absence of its own proteolytic complex in the raw materials in the technology the enzyme protosubtilin was used – the product of bacteria activity of Bacillus subtilis strain. It contains a complex of enzymes (neutral and alkaline proteinases, alpha-amylase, beta- glucanase, xylanase and cellulase), table salt, chemically precipitated chalk, corn flour. The choice of enzyme is also due to its low cost, compared to other enzymes (trypsin, pancreatin and others). The authors established the proteolytic activity of used for hydrolysis the enzyme preparation protosubtilin G3X – 560.77 µmol TYR/g. At the first stage of work the influence of temperature parameters on the intensity of the fermentation process is investigated. It was found that when the temperature rises to a certain level, the rate of hydrolysis increases, but then due to the thermal denaturation of the enzyme molecule, the activity of the latter decreases, which leads to a decrease in the speed of the process. It is experimentally established (Fig. 1) that the optimum temperature for enzymatic hydrolysis of BMW is (45±1)˚C. Figure 1. Dependence of the degree of hydrolysis on the temperature of the reaction environment. At the next stage of work, using the theory of experiment planning and computer modeling, a series of studies to optimize the stage of enzymatic hydrolysis of fish waste was carried out: it was specified the optimum amount of enzyme and fermentation duration at a constant temperature of 45˚С. Factors of optimization were: the concentration of the enzyme preparation– Х1 (% to the weight of raw materials) and the duration of the hydrolysis process – Х2 (hour). The value of the achieved hydrolysis degree – Y1 (%) was used as a response function (optimization parameter). Rotatable compositional plan for two factors provides for 9 experiments [22]. Limits and intervals of factors changes (Х1, Х2) are given in table 1. Table 1. The results of data processing Ser. No. Х1, % Х2, time У, % 1 0.6 2 23.50 2 0.6 5 23.90 3 1.5 2 26.5 4 1.5 5 27.0 5 1.05 3.5 27.50 http://www.iaeme.com/IJMET/index.asp 1885 editor@iaeme.com
  7. Developing the Technology and Evaluating the Biological Value of the Peptone From Secondary Products of Processing of Fish Raw Material of the Arctic Region 6 1.69 3.5 27.6 7 0.41 3.5 22.8 8 1.05 1.38 24.3 9 1.05 5.62 27.6 In accordance with the plan of the experiment, 9 variants of peptone samples were made according to the developed technological scheme. Mathematical processing of data obtained was performed using DataFit version 9.1 As a result of processing the following equation of regression of influence dependence of Х1 and Х2 factors on the generalized parameter of optimization is received: Y = a*x1+b*x12+c*x2+d*x22+e*x23 , (2) where the regression coefficients: а = 17,08; b = -6,42; с = 14,14; d = -3,80; e = 0.32 Fisher's criterion for this equation is 80.84, which means that with a given confidence probability (0.00044), the regression equation reliably describes the change in the optimization parameter from the influencing factors Х1 and Х2. The graphical interpretation of the regression equation is shown in figure 2. Figure 2. Graphical interpretation of the regression equation for determining the optimal parameters of the hydrolysis process To find the optimal values of the factors X1 and X2, which determine the optimal parameters of the hydrolysis process, the methods of mathematical processing (differentiation) were used. The values of these optimal factors are as follows: X1 (enzyme preparation concentration) - 1.33 % to the weight of raw material; X2 (duration of hydrolysis) - 3 hours. Thus, the parameters of enzymatic hydrolysis close to optimal were found. Taking into account the carried-out researches the technological scheme of pepton reception from bone-muscular waste of fish is developed (Fig.3). http://www.iaeme.com/IJMET/index.asp 1886 editor@iaeme.com
  8. Ludmila Kazimirovna Kuranova, Vlasimir Aleksandrovich Grokhovsky, Yulia Viacheslavovna Zhivlyantseva and Vasily Igorevich Volchenko bone-muscular waste cryogrinding adding water and enzyme preparation fermentation acidification to a pH of (4.5±0.1) separation of the dense part by centrifugation alkalinization to pH (8.1±0.1) separation of the dense portion of the filtration sublimation drying packaging Figure 3 Technological scheme of peptone production from bone-muscular waste. Frozen bone-muscular waste is crushed by cryoextrusion method and sent to hydrolysis. Hydrolysis is carried out in an aqueous medium (the ratio of minced meat and water is 1:1) with the help of an enzyme preparation protosubtilin of G3X brand with continuous stirring and at temperature of (45±1)˚С. After the hydrolysis is completed, the hydrolyzate is clarified in the acidic zone. To this end, the value of the hydrogen index is adjusted to the pH value (4.5±0.1) by concentrated hydrochloric acid and the mixture is heated to a temperature of 100˚С to inactivate the enzyme, and then it is sent to the separation of the dense part by centrifugation. The hydrolysate clarified in the acidic zone is directed to alkalinization with a concentrated solution of caustic soda to pH (8.1±0.1), heated to a temperature of 100˚С, after holding at this temperature for 15 minutes with stirring, directed to filtration. The liquid hydrolyzate is collected in a storage collector and transferred to freeze-drying. The dry product is collected in a hermetically sealed container and transferred to the packaging. The study of the quality of peptone obtained from bone-muscular cod waste (PBMW) on the optimized technological scheme. Pepton is an amorphous, fine powder of light beige color, odorless. The product is hygroscopic. Pepton has the ability to emulsify, foaming, when dissolved in water gives opalescent solutions, which confirms the preservation of the product properties of native protein. As a result of studies of the chemical composition, it was found that the content of protein substances in the obtained peptone is 92.3 %, the mass fraction of water is 4.7 %, the mass fraction of sodium chloride is 2.6 %, the mass fraction of fat is 0.3 %. Studies of the amino acid composition of the product found that PBMW proteins are characterized by a full set of protein amino acids, including essential ones (Fig. 4-6), Table. 2. The total amount of amino acids is 94.7 %, which is almost the same as the amount of protein substances in the product. Thus, all pepton protein can be considered as "true". http://www.iaeme.com/IJMET/index.asp 1887 editor@iaeme.com
  9. Developing the Technology and Evaluating the Biological Value of the Peptone From Secondary Products of Processing of Fish Raw Material of the Arctic Region Figure 4 Chromatogram characterizing the composition of amino acids in the obtained peptone sample: aspartic acid, glutamic acid, hydroxyproline, serine, glycine, histidine, arginine, threonine, alanine, proline, tyrosine, valine, isoleucine, leucine, phenylalanine, lysine Figure 5 Chromatogram obtained by high-performance liquid chromatography, revealing tryptophan in the fish peptone http://www.iaeme.com/IJMET/index.asp 1888 editor@iaeme.com
  10. Ludmila Kazimirovna Kuranova, Vlasimir Aleksandrovich Grokhovsky, Yulia Viacheslavovna Zhivlyantseva and Vasily Igorevich Volchenko Figure 6 Chromatogram obtained by HPLC, identifying in the composition of fish peptone sulphur-containing amino acids: cysteine, methionine Table 2 Amino acid composition and biological value of fish peptone proteins Amino-acid Content, mg/g Reference values [21], Amino acid score (AAS), of protein mg/g of protein % Tryptophan* 4.0 6.0 67 Lysine* 93.8 45.0 209 Histidine* 27.0 15.0 180 Threonine* 73.0 23.0 318 Cysteine 14.7 22.0 151 Methionine* 18.5 (methionine + cysteine) Valine* 44.3 39.0 114 Isoleucine* 42.3 30.0 141 Leucine* 73.8 59.0 125 Tyrosine 20.0 38.0 134 Phenylalanine* 31.0 (tyrosine + phenylalanine) Arginine 82.7 Aspartic acid 93.3 Serine 54.5 Glutamic acid 150.6 Proline 49.2 Oxyproline 17.9 Glycine 86.0 Alanine 82.7 Sum of essential amino 407.7 acids http://www.iaeme.com/IJMET/index.asp 1889 editor@iaeme.com
  11. Developing the Technology and Evaluating the Biological Value of the Peptone From Secondary Products of Processing of Fish Raw Material of the Arctic Region * )- essential amino acids In the minimum amount of peptone proteins there are tryptophan (4.0 mg/g of protein), in the maximum – hydroxyproline (150.6 mg/g of protein). The proteins of the studied peptone significantly contain such an essential amino acid as lysine, so the peptone can be recommended as additive for the enrichment of lysine-defective protein products, in particular plant ones. One gram of protein contains valine – 44.3 mg, isoleucine – 42.3 mg, leucine – 73.8 mg, lysine – 93.9 mg, methionine – 18.5 mg, tryptophan – 4.0 mg, threonine – 73.1 mg, phenylalanine – 31.0 mg, histidine – 93.3 mg . The quality of proteins was evaluated by the balance of their amino acid composition in comparison with the reference protein [21]. In peptone protein, the only limiting amino acid is tryptophan, score of which is 67% (Fig. 7). Figure 7 Content of essential amino acids in fish peptone and reference protein Fish peptone is characterized by a high content of amino acids, balance on seven essential amino acids and the presence of one limiting amino acid – tryptophan. The resulting protein product in terms of protein content and its properties corresponds to the category of protein isolates. Fish peptone can be recommended for food use as a protein supplement in food. 4. CONCLUSION In the course of the work the following results are obtained:  determined safety of bone-muscular cod waste;  developed and optimized the basic technological scheme of production of peptone (PBMW) from bone-muscular cod waste;  the following values of the optimal factors of the hydrolysis process are established: hydrolysis temperature - (45±1)˚С, duration - 3 hours, the http://www.iaeme.com/IJMET/index.asp 1890 editor@iaeme.com
  12. Ludmila Kazimirovna Kuranova, Vlasimir Aleksandrovich Grokhovsky, Yulia Viacheslavovna Zhivlyantseva and Vasily Igorevich Volchenko concentration of the enzyme preparation protosubtilin G3X with activity 560.77 mmol TYR/g -1.33 % to the weight of raw materials;  chemical and biochemical parameters of PBMW quality are investigated, its biological value is defined;  PBMW can be recommended for use as a full-fledged protein component of food products. FUNDING STATEMENT The work was supported by the Ministry of education and science of the Russian Federation, project 15.11168.2017/8.9. REFERENCES [1] Hambræus, L. Protein and Amino Acids in Human Nutrition: Reference Module in Biomedical Sciences, 2014. DOI 10.1016/B978-0-12-801238-3.00028-3 [2] Maximizing the value of marine by-products. In Shahidi, F., ed., CRC Press: Boca Raton, Boston, New York, Washington DC, 2007, pp. 375. ISBN: 978-1-84569-013-7 [3] Vysotsky, V. G., Yatsyshina, T. A. and Mamaev E.M. On the question of terminology used in biological characterization of protein quality. Nutrition, 6, 1977, pp. 3–9. [4] Orlova, T. A. and Zenzerov, V. S. Technologies of reception products and biologically active substances from marine hydrobionts. Apatity: Publishing house of the KSC RAS, 2004, pp. 277. [5] Mukhin, V. A. and Novikov, V. Y. Enzymatic protein hydrolysates of tissues of marine hydrobionts: obtaining, properties and practical use. Murmansk: Publishing house of PINRO, 2001, pp. 101. [6] Kristinsson, Hordur G. and Rasco, Barbara A. Fish Protein Hydrolysates: Production, Biochemical, and Functional Properties. Critical Reviews in Food Science and Nutrition, 40(1), 2010, pp. 43–81. http://dx.doi.org/10.1080/10408690091189266 [7] Katysheva, E. G. The role of the Northern Sea Route in Russian LNG Projects Development. IOP Conference Series: Earth and Environmental Science, 180(1), 2018, article 012008. DOI: 10.1088/1755-1315/180/1/012008 [8] Strizhenok, A., Korelskiy, D. Assessment of the state of soil-vegetation complexes exposed to powder-gas emissions of nonferrous metallurgy enterprises. Journal of Ecological Engineering, 17(4), 2016, pp. 25–29. DOI: https://doi.org/10.12911/22998993/64562 [9] Technochemical properties of commercial fish in the North Atlantic and adjacent seas of the Arctic ocean. In Troyanovsky, F. M., ed., Murmansk: Publishing house of PINRO, 1997, pp. 183. [10] Tutelian, V. A. Chemical composition and caloric content of Russian food products: Information guide. Moscow: DeLi plus, 2012, pp. 284. ISBN: 978-5-905170-20-1 [11] Maksimova, E. M. Development of technology for utilization of protein waste by enzymatic hydrolysis. Bulletin of the Murmansk State Technical University. Murmansk, 9(5), 2006, pp. 875–879. https://cyberleninka.ru/article/v/razrabotka-tehnologii- utilizatsii-belkovyh-othodov-metodom-fermentativnogo-gidroliza [12] Mezenova, O. Ya. and Zemlyakova, E. S. Basic principles of secondary fish raw materials processing for food bioproducts. KSTU News, 35, 2014, pp. 120–130. [13] Zhivliantseva, Yu. V. and Kuranova, K. Assessment of the suitability of bone-muscular waste for the production of cod protein hydrolysate. Proceedings X Russian National Scientific and Practical Conference of young scientists on the problems of aquatic http://www.iaeme.com/IJMET/index.asp 1891 editor@iaeme.com
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