Degree Of Hydrolysis Research Articles

Recombinant expression and characterization of the endochitinase Chit36-TA from Trichoderma asperellum in Komagataella phaffii for chitin degradation of black soldier fly exuviae

The natural polymer chitin is an abundant source for valuable N-acetylchitooligosaccharides and N-acetylglucosamine applicable in several industries. The endochitinase Chit36-TA from Trichoderma asperellum was recombinantly expressed in Komagataella phaffii for the enzymatic degradation of chitin from unused insect exuviae into N-acetylchitooligosaccharides. Chit36-TA was purified by Ni–NTA affinity chromatography and subsequently biochemically characterized. After deglycosylation, the endochitinase had a molecular weight of 36 kDa. The optimum pH for Chit36-TA was 4.5. The temperature maximum of Chit36-TA was determined to be 50 °C, while it maintained > 93% activity up to 60 °C. The chitinase was thermostable up to 45 °C and exhibited ~ 50% activity after a 15 min incubation at 57 °C. Chit36-TA had a maximum specific enzyme activity of 50 nkat/mg with a Km value of 289 µM with 4-methylumbelliferyl-N,N′,N″-triacetyl-β-chitotrioside as substrate. Most tested cations, organic solvents and reagents were well-tolerated by the endochitinase, except for SDS (1 mM), Cu2+ (10 mM) and Mn2+ (10 mM), which had stronger inhibitory effects with residual activities of 3, 41 and 28%, respectively. With a degree of hydrolysis of 32% applying colloidal shrimp chitin (1% (w/v)) and 12% on insect larvae (1% (w/v)) after 24 h, the endochitinase was found to be suitable for the conversion of colloidal chitin as well as chitin from black soldier fly larvae into water-soluble N-acetylchitooligosaccharides. To prove scalability, a bioreactor process was developed in which a 55-fold higher enzyme activity of 49 µkat/l and a tenfold higher protein expression of 1258 mg/l were achieved.

Regulating antigenicity of α‐lactalbumin based on enzymolysis: Insights into structure and linear epitopes

This study investigated the effect of enzymatic treatment on the allergenicity of α‐lactalbumin (ALA). Utilising the BIOPEP database, we identified five proteases that optimally clear allergen epitopes, while considering cost‐efficiency. Notably, alkaline protease demonstrated the highest antigen reduction rate of 46.46%, with a hydrolysis degree of 30.01%, and 87.80% of the resulting peptides were found to be smaller than 1 kDa. Peptidomics analysis revealed AA32–38 and AA42–48 as key linear epitopes of ALA. Furthermore, alkaline protease treatment significantly reduced the proportion of hydrophilic amino acids, thereby decreasing the allergen potential of ALA by lowering its IgE/IgG binding affinity.

Antioxidant and Anti-Atherosclerosis Activities of Hydrolyzed Jellyfish Collagen and Its Conjugate with Black Jelly Mushroom Extract.

Atherosclerosis, a noncommunicable disease caused by cholesterol plaque, can cause chronic diseases. The antiplatelet medicines used in its treatment can cause complications. Marine collagen peptides can be used as a natural atherosclerosis remedy. The present study investigated the preparation and characterization of hydrolyzed collagen (HC) from jellyfish and its conjugation with black jelly mushroom extract (BJME). Their cytotoxicity and ability to prevent cholesterol-induced endothelial cell injury were also examined. HC was prepared using Alcalase or papain hydrolysis (0.2-0.4 units/g of dry matter (DM)). Higher yield, degree of hydrolysis, and antioxidant activities (AAs) were found in the HC obtained from Alcalase, especially at 0.4 units/g DM (A-0.4), compared to other processes (p < 0.05). Thus, A-0.4 was further conjugated with BJME (1-4%, w/w of HC). The HC-2%BJME conjugate showed the highest surface hydrophobicity and AAs compared to other samples. The FTIR spectra and size distribution also confirmed the conjugation between HC and BJME. When EA.hy926 endothelial cells were treated with HC or HC-2%BJME (25-1000 µg/mL), HC-2%BJME had no cytotoxicity, whereas HC at 1000 µg/mL induced cytotoxicity (p < 0.05). Both samples also exhibited protective ability against cholesterol-induced apoptosis and VE-cadherin downregulation of cells. Therefore, HC and conjugate could be natural agents for preventing atherosclerosis.

Preparation and characterization of silica sol-impregnated glass fibers

Abstract Silica sol-impregnated glass fiber products were prepared via the sol-gel method, with tetraethoxysilane (TEOS) as a precursor and hydrochloric acid (HCl) as a catalyst, and characterized by 1H NMR, FT-IR, and solid 29Si NMR analyses. The results indicated that the structure of the impregnating agents was significantly affected by the amount of water added. With a decrease in the molar ratio of added water, the degree of TEOS hydrolysis was reduced, and more ethoxy groups -OCH2CH3 were formed, resulting in the formation of sol products with a linear structure. Furthermore, contact angle and water absorption tests showed that as the molar ratio of water increased, the water contact angle decreased, whereas the water absorption of impregnated glass fiber increased. These results indicate that a higher hydrolysis degree of TEOS with more added water generates more Si-OH groups, facilitating the cross-linking of sol products to form a network structure.

Characterization and application of Bacillus velezensis D6 co-producing α-amylase and protease.

Research on the co-production of multiple enzymes by Bacillus velezensis as a novel species is still a topic that needs to be studied. This study aimed to investigate the fermentation characteristics of B. velezensis D6 co-producing α-amylase and protease and to explore their enzymatic properties and applications in fermentation. The maximum co-production of α-amylase and protease reached 13.13 ± 0.72 and 2106.63 ± 64.42 U mL-1, respectively, under the optimal fermented conditions (nutrients: 20.0 g L-1 urea, 20.0 g L-1 glucose, 0.7 g L-1 MnCl2; incubation conditions: initial pH 7.0, temperature 41 °C, 8% inoculation size and 30% working volume). Moreover, the genetic co-expression of α-amylase and protease increased from 0 to 24 h and then decreased after 36 h at the transcriptional level, which coincided with the growth trend of B. velezensis D6. The optimal reaction temperature of α-amylase was 55-60 °C, while that of protease was 35-40 °C. The activities of α-amylase and protease were retained by over 80% after thermal treatment (90 °C, 1 h), which indicated that two enzymes co-produced by B. velezensis D6 demonstrated excellent thermal stability. Moreover, the two enzymes were stable over a wide pH range (pH 4.0-8.0 for α-amylase; pH 4.0-9.0 for protease). Finally, the degrees of hydrolysis of corn, rice, sorghum and soybeans by α-amylase from B. velezensis D6 reached 44.95 ± 2.95%, 57.16 ± 2.75%, 52.53 ± 4.01% and 20.53 ± 2.42%, respectively, suggesting an excellent hydrolysis effect on starchy raw materials. The hydrolysis degrees of mackerel heads and soybeans by protease were 43.93 ± 2.19% and 26.38 ± 1.72%, respectively, which suggested that the protease from B. velezensis D6 preferentially hydrolyzed animal-based protein. This is a systematic study on the co-production of α-amylase and protease by B. velezensis D6, which is crucial in widening the understanding of this species co-producing multi-enzymes and in exploring its potential application. © 2024 Society of Chemical Industry.

Physicochemical properties, antioxidant and antidiabetic activities of different hydrolysates of goat milk protein

There is growing interest in the origin, preparation, and application of bioactive peptides. This study investigated the impact of 6 enzymes on the structural, physicochemical properties, antioxidant activities, and antidiabetic potential of defatted fresh goat milk. Structural and functional changes resulting from enzymatic hydrolysis were assessed using gel electrophoresis, laser particle size analysis, multi-spectroscopy, and evaluations of foaming and emulsification properties. Antioxidant capacity was determined through free radical scavenging, Fe2+ chelation, and reducing ability experiments. Additionally, the inhibitory effects of the hydrolysates on α-glucosidase and α-amylase were measured to evaluate antidiabetic activity. Results showed that enzymatic hydrolysis disrupted the spatial structure of goat milk protein and reduced its molecular weight. Papain hydrolysate exhibited the highest degree of hydrolysis (32.87 ± 0.11%) and smallest particle size (294.75 ± 3.33 nm), followed by alcalase hydrolysate (29.12 ± 0.09%, 302.03 ± 7.28 nm). Alcalase hydrolysate showed the best foaming properties, while papain hydrolysate demonstrated the strongest DPPH and hydroxyl radical scavenging activity, Fe2+ chelation, and antidiabetic potential. These findings provide solid theoretical basis for utilizing defatted goat milk as functional ingredients or excipients in the food, medical, and cosmetic industries.

Ultra-high pressure homogenization does not impair the in vitro digestibility of egg yolk granule proteins

Egg yolk granule is a promising ingredient in the food industry. Recent studies have shown that ultra-high pressure homogenization (UHPH) improved the techno-functional properties of the granule, but they did not evaluate its impact on digestibility. Consequently, this work aimed to evaluate the impact of UHPH-treatment (300 MPa and 4 passes) on granule digestibility by using a static in vitro digestion method. While UHPH destabilized the high molecular weight granule lipoproteins, the treatment did not affect the in vitro digestibility of egg yolk granule since similar protein/peptide profiles were obtained from gel electrophoresis and RP-HPLC analysis. Moreover, the degree of hydrolysis was similar between control and UHPH-treated granules with values ranging from 11.5 to 25.2% for the gastric and intestinal phases, respectively. Finally, apovitellin 3-4, apovitellenin IV and phosvitin were specifically digested during the gastric and intestinal phases. Consequently, modifications induced by UHPH on egg yolk granule were not correlated with in vitro digestion experiments. Nevertheless, this work represents an interesting added-value to support the development of innovative egg yolk granule-based ingredients for the food industry.

Ultrasound and enzyme treatments on morphology, structures, and adsorption properties of cassava starch

Porous starch materials are environmentally friendly and renewable and exhibit high adsorption performances. Ultrasound and compound enzyme (α-amylase and glucoamylase) treatments were applied to prepare modified cassava starch. The granules, crystal morphology, crystal structure, and molecular structure of starch were investigated. The hydrolysis degree, solubility, swelling, and adsorption properties of cassava starch were analyzed. After the cassava starch was modified by ultrasound and enzyme treatments, the granule size of the starch decreased, and the surfaces were eroded to form pits, grooves and cavity structure. The starch spherulites weakened or even disappeared. The functional groups of starch did not change significantly, but the degree of crystal order decreased. The double-helix structure was reduced, and the crystal structure was composed of A + V-type crystals, with a decrease in crystallinity. The gelatinization temperature and thermal degradation temperatures enhanced. The enzymatic hydrolysis degree and solubility of the modified cassava starch increased. The swelling degree decreased, and oil adsorption, water adsorption improved. MB adsorption behavior of modified cassava starch closely followed a pseudo-second-order kinetics model and the Langmuir isotherm equation. These findings could help to understand the relationship between the structure and properties of modified starch, and guide its application in the field of adsorption.

Protein Hydrolysates from Salmon Heads and Cape Hake By-Products: Comparing Enzymatic Method with Subcritical Water Extraction on Bioactivity Properties.

Fish by-products can be converted into high-value-added products like fish protein hydrolysates (FPHs), which have high nutritional value and are rich in bioactive peptides with health benefits. This study aims to characterise FPHs derived from salmon heads (HPSs) and Cape hake trimmings (HPHs) using Alcalase for enzymatic hydrolysis and Subcritical Water Hydrolysis (SWH) as an alternative method. All hydrolysates demonstrated high protein content (70.4-88.7%), with the degree of hydrolysis (DH) ranging from 10.7 to 36.4%. The peptide profile of FPHs indicated the breakdown of proteins into small peptides. HPSs showed higher levels of glycine and proline, while HPHs had higher concentrations of glutamic acid, leucine, threonine, and phenylalanine. Similar elemental profiles were observed in both HPHs and HPSs, and the levels of Cd, Pb, and Hg were well below the legislated limits. Hydrolysates do not have a negative effect on cell metabolism and contribute to cell growth. HPSs and HPHs exhibited high 2,2'-azino-bis(3 ethylbenzthiazoline-6)-sulfonic acid (ABTS) radical scavenging activity, Cu2+ and Fe2+ chelating activities, and angiotensin-converting enzyme (ACE) inhibitory activity, with HPHs generally displaying higher activities. The α-amylase inhibition of both FPHs was relatively low. These results indicate that HPHs are a promising natural source of nutritional compounds and bioactive peptides, making them potential candidates for use as an ingredient in new food products or nutraceuticals. SWH at 250 °C is a viable alternative to enzymatic methods for producing FPHs from salmon heads with high antioxidant and chelating properties.

Biocontrol Strategy of Listeria monocytogenes in Ready-to-Eat Pork Cooked Ham Using Peptic Hydrolysates of Porcine Hemoglobin.

Listeria monocytogenes is a foodborne pathogen that represents a serious concern for ready-to-eat (RTE) meat products due to its persistence in production facilities. Among the different strategies for the control of this pathogen, the use of antimicrobial peptides derived from food by-products, such as slaughterhouse blood proteins, has emerged as a promising biocontrol strategy. This study evaluated for the first time the use of peptic hydrolysates of porcine hemoglobin as a biocontrol strategy of L. monocytogenes in RTE pork cooked ham. Pure porcine hemoglobin (Hb-P) and porcine cruor (P-Cru) were hydrolyzed using pepsin at different temperatures (37 °C for Hb-P and 23 °C for P-Cru) for 3 h. Then, the hydrolysates were characterized in terms of their degree of hydrolysis (DH), peptide population, color, and antimicrobial activity (in vitro and in situ) against three different serotypes of L. monocytogenes. Reducing the hydrolysis temperature of P-Cru by 14 °C resulted in a 2 percentage unit decrease in DH and some differences in the peptide composition. Nevertheless, the antimicrobial activity (in situ) was not significantly impacted, decreasing the viable count of L. monocytogenes by ~1-log and retarding their growth for 21 days at 4 °C. Although the color of the product was visibly altered, leading to more saturated reddish and yellowish tones and reduced brightness, the discoloration of the hydrolysates can be addressed. This biopreservation approach holds promise for other meat products and contributes to the circular economy concept of the meat industry by valorizing slaughterhouse blood and producing new antilisterial compounds.

Role of the intestines on the muscle quality of Pacific white shrimp (Litopenaeus vannamei) during chilled storage: Physicochemical and label-free–based peptidomics analyses

The effect of shrimp deveining on the quality of Pacific white shrimp muscle was investigated by analyzing the protein degradation during chilled storage via physicochemical and label-free peptidomics analyses. In this study, shrimp with intact intestines were in the control group (CS), while deveined shrimp (DS) were in the treatment group. The total viability count (TVC), total volatile base nitrogen (TVB-N) content, and trichloroacetic acid (TCA)-soluble peptide content in all of the shrimp groups gradually increased with prolonged chilled storage. However, in the later stages of chilled storage, the DS samples exhibited significantly lower TVB-N, total bacterial, and TCA-soluble peptide contents than the CS samples, indicating that deveining treatment effectively prolonged shrimp quality. The peptidomics analysis revealed varying degrees of protein hydrolysis in the DS and CS samples during chilled storage. A total of 396 differentially abundant peptides (DAPs) were identified in the DS compared with the CS, comprising 98 upregulated and 298 downregulated segments. This suggests that the removal of the intestine effectively inhibits protein hydrolysis. Gene ontology (GO) analysis suggested that the DAPs were mainly involved in catalytic activity, binding, and metabolic processes. The cluster of orthologous groups of protein (COG) analysis showed that the cytoskeleton dynamics of the muscle proteins underwent considerable alterations influenced by the shrimp's intestines during chilled storage.

Thermally Stabilised Poly(vinyl alcohol) Nanofibrous Materials Produced by Scalable Electrospinning: Applications in Tissue Engineering.

Electrospinning is a widely employed manufacturing platform for tissue engineering applications because it produces structures that closely mimic the extracellular matrix. Herein, we demonstrate the potential of poly(vinyl alcohol) (PVA) electrospun nanofibers as scaffolds for tissue engineering. Nanofibers were created by needleless direct current electrospinning from PVA with two different degrees of hydrolysis (DH), namely 98% and 99% and subsequently heat treated at 180 °C for up to 16 h to render them insoluble in aqueous environments without the use of toxic cross-linking agents. Despite the small differences in the PVA chemical structure, the changes in the material properties were substantial. The higher degree of hydrolysis resulted in non-woven supports with thinner fibres (285 ± 81 nm c.f. 399 ± 153 nm) that were mechanically stronger by 62% (±11%) and almost twice as more crystalline than those from 98% hydrolysed PVA. Although prolonged heat treatment (16 h) did not influence fibre morphology, it reduced the crystallinity and tensile strength for both sets of materials. All samples demonstrated a lack or very low degree of haemolysis (<5%), and there were no notable changes in their anticoagulant activity (≤3%). Thrombus formation, on the other hand, increased by 82% (±18%) for the 98% hydrolysed samples and by 71% (±10%) for the 99% hydrolysed samples, with heat treatment up to 16 h, as a direct consequence of the preservation of the fibrous morphology. 3T3 mouse fibroblasts showed the best proliferation on scaffolds that were thermally stabilised for 4 and 8 h. Overall these scaffolds show potential as 'greener' alternatives to other electrospun tissue engineering materials, especially in cases where they may be used as delivery vectors for heat tolerant additives.

Hydrolysis Optimization of Thraustochytrid (Heterotrophic Protist) Biomass for the Production of a New and Sustainable Ingredient for the Aquafeed Industry

Thraustochytrids biomass, rich in docosahexaenoic acid (DHA), could be a sustainable ingredient for the aquafeed industry. However, its production cost renders it less competitive than fishmeal and fish oil from wild-caught fish. This study proposes optimizing the hydrolysis conditions of thraustochytrid biomass to generate an ingredient with improved properties thanks to the production of peptides with different biological activities. The improved nutritional value of the biomass could justify its use to decrease the amount of wild-caught fish in aquafeed, which would enhance the sustainability of fish aquaculture. First, two commercial proteases, Papain and Alcalase 2.4L, were compared for their capacity to hydrolyze the biomass. The best degree of hydrolysis (19 ± 1%) was obtained with Alcalase 2.4L, which was then used for the optimization of the pH, enzyme concentration and reaction time using response surface methodology. The results showed that the highest concentration of peptides and DHA in the aqueous phase was obtained with a pH, enzyme concentration and reaction time of 7.5, 2.7% and 205 min, respectively. If thraustochytrid hydrolysates prove to improve fish fitness, this mild and free-solvent process protocol could be used to produce a sustainable ingredient for aquafeed.

Valorisation of Tenebrio molitor beetles: enzymes-based bioprocessing to obtain a highly digestible protein hydrolysate

Abstract The biological valorisation of edible insects is an eco-saving approach that may increase their use for food and feeding purposes. Beetles of T. molitor are a direct waste of insect farms since their protein have a low digestibility. Currently, the degree of hydrolysis achieved with biological treatments are much lower than with chemical treatments. This work focused on the enzymatic hydrolysis of protein from T. molitor beetles using proteases. The obtained protein hydrolysate was evaluated regarding the digestibility and functional properties. Higher hydrolysis degree (HD; 47%) was obtained using a protease from Aspergillus oryzae, which efficiently separated protein from chitin. A full factorial design determined that a liquid:solid ratio (LSR) of 15 and an enzyme load of 3200 U/g increase HD up to 78%. The protein hydrolysate showed higher digestibility (77.3%) compared to beetles’ meal before (17.8%) and after defatting (25.7%). The protein hydrolysate showed a high antioxidant activity (462.1 ± 7.48 μmol TE/g DW), it was highly soluble at pH ranging between 3 and 9 and showed an emulsification activity and stability of 53.6% and 82.2%, respectively. Therefore, this bioprocess improved the use of beetles, obtaining protein with greater digestibility to be used as food.

Functional improvement and characterization of protein hydrolysates prepared by the fermentation of irradiated tilapia skin

Gamma irradiation (γ-irradiation) sterilizes and modifies proteins. This study aimed to enhance the performance of protein hydrolysates of fermented tilapia fish skin using irradiation. The results showed that γ-irradiation could effectively kill harmful microorganisms in fish skin at 6 kGy. Irradiation at 1–6 kGy dose disintegrated the internal structure of fish skin proteins, thus increasing the degree of hydrolysis after fermentation by Bacillus subtilis L4; however, irradiation at 9–12 kGy caused cross-linking and aggregation of fish skin proteins. The solubility, and water and oil holding capacity of the protein hydrolysates were significantly improved compared to those of the non-irradiated sample. The maximum emulsifying activity (164.61±16.28 m2/g), foaming activity (91.13±2.30 mL), and surface hydrophobicity index (5766.43±770.56) were observed in the protein hydrolysates fermented from fish skin irradiated at 12 kGy. The antioxidant activity of the protein hydrolysates increased with increasing irradiation doses. This suggested that the functional properties and antioxidant activity of protein hydrolysates from fish skin fermentation can be improved by appropriate irradiation pretreatment, providing a feasible method for improving the utilization and quality of proteins in fishery waste.

Controlled and rapid microreactor-assisted hydrolysis of sesame protein by a novel nanobiocatalyst; immobilized alcalase over activated sol–gel magnetic nanosupport

In this study, sesame cake was valorized to produce sesame protein hydrolysate (SPH) with improved functional and antioxidant properties using a recoverable nanomagnetic immobilized Alcalase (ALC); the process was accelerated in the microreactor through a controlled-hydrolysis technique. For this purpose, silica-coated magnetic nanoparticles (Fe3O4@SiO2) were synthesized and functionalized with tetraethoxysilane (TEOS), N-(trimethoxysilylpropyl) ethylenediamine (TMSED), and finally activated with glutaraldehyde (GLU) to produce Fe3O4@SiO2-TMSED-GLU. Immobilization efficiency of ALC was 88.78 % in the optimum conditions of ALC concentration (1.23 mg/mL), immobilization period (119.12 min), and temperature (38.93 °C). The successful synthesis of Fe3O4@SiO2-TMSED-GLU-ALC magnetic nanobiocomposite (MNBC) was approved using FTIR, XRD, VSM, FESEM, EDX, and elemental mapping. The effective immobilisation of the enzyme was responsible for the decrease in the intensity of Fe3O4 diffraction peaks (in XRD) in MNBC and a reduction in saturation magnetization (in VSM) following ALC immobilisation. Microreactor-intensified hydrolysis in the optimized conditions of MNBC = 4.85 wt%, reaction time = 54.10 s, and temperature = 46.95 °C, led to a degree of hydrolysis = 7.19 %. After 10 hydrolysis cycles, MNBC had relative activity > 80 %, due to the appropriate covalent bonding between the enzyme and the support and the short duration of the process. The produced SPH had superior foaming capacity (113.09 %) compared to sesame protein (76.34 %). The emulsion activity and emulsion stability of SPH were 28.78 m2/g and 56.43 min, respectively, which were higher than those of the sesame protein.

Optimization of the autolysis of rainbow trout viscera for amino acid release using response surface methodology

Background Due to the huge amounts of their production in Europe, their environmental impact, and the difficulty in processing them, there is a clear necessity for the valorization of rainbow trout viscera. Considering that the production of fishmeal with viscera can be problematic, and in order to make viscera more profitable, the production of fish protein hydrolysates has been considered. Although silage and enzymatic hydrolysis are the most common methods for obtaining hydrolysates, autolysis has emerged as an alternative method that uses endogenous enzymes of the viscera. Methods Considering the stability and characteristics of the enzymes, a factorial design was carried out using three variables: pH, temperature, and water content. The design resulted in 15 experiments, and the results were analyzed using response surface methodology. The optimum parameters were validated by comparing the predicted outcomes with experimental results. Additionally, a kinetics study was conducted to shorten the autolysis time. Results from autolysis were compared with those from silage and enzymatic hydrolysis in a previous study. Results The optimal conditions for achieving the highest degree of hydrolysis and yield of free amino acids (FAAs) per 100 g of viscera and per total protein were determined to be a pH of 8, a temperature of 40 ºC, and a water content of 6.85%. The pH and content of the added water were found to be significant variables during autolysis (p &lt; 0.05). The kinetic study showed that 7 h was still required to be effective. Conclusions Autolysis achieved a lower degree of hydrolysis than silage; however, as it solubilized more protein, the global yield of free amino acids per 100 g of viscera was slightly higher. It was concluded that endogenous alkaline proteases could be used in an autolytic process to obtain a free amino acid-rich hydrolysate from trout viscera.

Valorization of Saithe (Pollachius virens) Residuals into Protein Hydrolysates-Silaging as Preservation Technology.

Silaging can be used as preservation technology to valorize currently discarded raw material into protein hydrolysate on board deep-sea vessels. The aim of this study was to investigate the effect of sorting and raw material freshness on the quality and yield of protein hydrolysates obtained through silaging of saithe (Pollachius virens) viscera. Additionally, the effect of using acid-containing antioxidants was tested. Out sorting of the liver prior to silaging resulted in slightly higher hydrolysate yields. The hydrolysates with the highest protein contents were obtained from silages made from fresh raw materials (day 0), and the content decreased significantly after longer storage of the raw material (2-3 days at 4 °C). Storage of the raw material for 1 day did not affect the quality. However, a significantly higher degree of hydrolysis (DH), content of free amino acids (FAA), and total volatile basic nitrogen (TVB-N) were obtained when raw materials were stored for 3 days. The FAA composition was influenced by the raw material's freshness, with increases in free glutamic acid and lysine and a decrease in free glutamine after longer storage. None of the studied parameters were significantly affected by out sorting of liver or the addition of antioxidants. Overall, the results indicate that the whole fraction of the viscera can be utilized without reducing the quality of the hydrolysate and that the raw material should be stored for a maximum of 1 day prior to preservation to optimize the quality.

Изучение некоторых функциональных характеристик ферментативных гидролизатов пищевых белков

Food allergy is a matter of global concern, proteins being a popular allergen worldwide. Enzymatic protein hydrolysates serve as hypoallergenic components in functional foods. This article describes enzymatic hydrolysates of egg and soy proteins as potential antigens, sensitizers, and antioxidants. The research featured soy protein isolate and egg albumin. The protein hydrolysates were obtained in a two-step process with pepsin and trypsin to measure hydrolysis, total antioxidant activity, and residual antigenicity. The sensitizing ability of native proteins and their hydrolysates was studied using a delayed-type hypersensitivity (DTH) skin test of paw swelling in mice. The egg albumin and soy protein were hydrolyzed by sequentially adding the proteases, i.e., pepsin and trypsin. The degree of protein hydrolysis was calculated against total nitrogen and its contents in non-hydrolyzed raw materials and hydrolysate. It was 82.6% for the egg hydrolysate and 88.3% for the soy hydrolysate, the total antioxidant activity being 114.3 and 91.4 mg/100 mL, respectively. The residual antigenicity of the hydrolysates determined by ELISA in a sandwich version was 1.55×10–4 and 3.30×10–4 RU, respectively. The native egg and soy proteins demonstrated good allergenic properties, and the DTH response index increased fivefold. The hydrolysates reduced the DTH response indices by an average of 3.5 for egg hydrolysate and 2.6 times for soy hydrolysate. The resulting enzymatic hydrolysates of egg albumin and soy protein demonstrated a high degree of hydrolysis and antioxidant activity. They had low residual antigenicity (10–4) and low sensitizing ability. The resulting enzymatic hydrolysates could be used as hypoallergenic components and antioxidants in new functional foods.

Comparative studies on composition of feed additives for juvenile fish produced with bioconversion of fish raw materials

Technology of feed additive for fry feeding in fish farms is developed to provide both important nutrients (proteins, lipids, etc.) and optimal peptide composition of protein that is achieved using proteolysis of raw materials under certain conditions. For this purpose, enzymatic hydrolysis of substandard small-sized frozen pollock was used. The rational duration of the hydrolysis process was 60 min. that provided the degree of protein hydrolysis of 26–28 %. The experimental fermentolysate was concentrated to a water content of 3–4 % and presented as a feed additive TINRO-60. This product is compared with other feed additives (dried fermentolysates) produced in Russia from fish raw materials using bioconversion, such as Biomarin, Bioprom and Vakhob. General chemical composition of tested products was determined by standard methods of analytical chemistry; composition and content of fatty acids — by gas-liquid chromatography; microand macroelements, as well as toxic metals — by atomic absorption spectrophotometry; fractional composition of proteins and peptides — by high pressure gel permeation chromatography; amino acid composition — by ion exchange chromatography on a high-speed analyzer. All tested samples contain similar amount of the main feed component — protein substances. The quantitative ratio of peptide fractions is also comparable, the sum of the most active fractions (MW 0.1−10.0 kDa) in TINRO-60 and Biomarin is close to their level in tissues of Artemia, natural food for fish fry. The content of essential amino acids, EPA and DHA is significant in Biomarin, TINRO-60 and Bioprom, but almost twice lower in Vakhob. Analysis of mineral composition showed that bioconversion into dry fermentolysates does not cause concentration of toxic elements. High quality of the protein feed additive TINRO-60 produced by bioconversion of nonstandard fish raw materials under certain conditions is proved.