Preparation Of Hydrolysates Research Articles

Food utility potential of protease obtained from insect‐commensal Bacillus subtilis AU‐2

AbstractBACKGROUNDBacillus subtilis AU‐2, isolated from the gut of Tribolium castaneum, was used for protease production. The purified protease was evaluated for its potential in food‐related applications including meat tenderization, milk coagulation, and the preparation of enzymatic soybean hydrolysates. Enzymatic hydrolysis of soy protein is an effective method for producing protein hydrolysates with optimal techno‐functional properties.RESULTSThis study confirmed that B. subtilis AU‐2 is a commensal with T. castaneum, within the insect gut flora. The purified protease obtained from B. subtilis AU‐2 exhibited meat tenderization activity and an ability to promote milk coagulation within a 20 min timeframe. Soybean hydrolysate prepared using the protease exhibited a relatively higher degree of hydrolysis (20.1%) than pepsin, trypsin, and papain. The sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) peptide fragmentation pattern of the enzymatically prepared soy protein hydrolysate revealed the disappearance of ~35 kDa protein bands after 4 h protease treatment. The prepared soy protein hydrolysate showed improved emulsifying capacity (EC) (705 mL g−1) and exhibited significant 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) antioxidant activity (77.89%) and hydroxyl (OH) radical scavenging activity (97.23%).CONCLUSIONSThe protease obtained from the newly isolated B. subtilis AU‐2 strain exhibited potential for food‐related applications, including meat tenderization, milk clotting, and soybean hydrolysate preparation. Its accessibility opens avenues for utilization in the food processing industry. © 2024 Society of Chemical Industry.

Preparation and Characterization of Calcium-Chelated Sea Cucumber Ovum Hydrolysate and the Inhibitory Effect on α-Amylase.

α-amylase can effectively inhibit the activity of digestive enzymes and alter nutrient absorption. The impact of ovum hydrolysates of sea cucumbers on α-amylase activity was investigated in this study. The protein hydrolysates generated using different proteases (pepsin, trypsin, and neutral protease) and molecular weights (less than 3000 and more than 3000) were investigated. The results showed that all three different hydrolysates demonstrated calcium-chelating activity and induced a fluorescence-quenching effect on α-amylase. The sea cucumber ovum hydrolysate with a molecular weight of less than 3000 Da, isolated using trypsin, showed the most effective inhibitory effect on α-amylase, with an inhibition rate of 53.9%, and the inhibition type was identified as mixed forms of inhibition. In conclusion, the generation and utilization of protein hydrolysates from sea cucumber ovum as a functional food ingredient could be a potential approach to add value to low-cost seafood by-products.

Optimization of Hydrolysis Conditions and Bio-functional Properties of Whey Protein Hydrolysate Obtained from Ash Gourd (Benincasa hispida) Protease

Background: Enzymatic hydrolysis alters the bio-functionality of whey proteins by generating bioactive peptides. Plant serine proteases as an alternative to animal and microbial proteases, recently considered useful for whey protein hydrolysate preparation to enhance nutritional and bio-functional properties. Objectives: The present study thus aims at optimization of whey protein hydrolyzing conditions employing ash gourd protease (AGP) and evaluation of 2, 2 diphenyl -1-picrylhydrazyl (DPPH) radical scavenging (antioxidant) and angiotensin I- converting enzyme (ACE) inhibition activities and potency of the generated hydrolysate. Methods: Optimization was accomplished using response surface methodology and generating treatment combinations applying central composite design in two steps: first temperature (50-90°C) and pH (6-9) was optimized adding same concentration of AGP (0.3% v/v) at reconstituted WPC (0.6% protein) and then E/S ratio (2-6 %v/v) and hydrolysis time (1-6h) was optimized at reconstituted WPC (4% protein). The effect of hydrolysis time at optimized condition was also evaluated with response to degree of hydrolysis, antioxidant and ACE inhibition activities of the generated hydrolysate and its ultra-centrifuged fractions ≤3kDa and >3kDa. The AGP (Adjusted to activity 2U/mL) obtained after partially purification by three phase partitioning method was used for hydrolysis experiments. Results: The optimum conditions for enzymatic hydrolysis of whey protein to achieve maximum degree of hydrolysis (13.99%) and DPPH radical scavenging activity (23.05%) were observed at 7.48 and 66.40°C. Similarly, the enzyme performed optimally at enzyme by substrate ratio of 5.85% (v/v) for 6 hours of hydrolysis, providing 19.31 % of degree of hydrolysis and 47.71 % of DPPH radical scavenging activity. AGP hydrolyzed whey protein compared to un-hydrolysed whey protein had significantly higher degree of hydrolysis, DPPH radical scavenging and ACE inhibition activity. The lower molecular weight peptides fractions ≤3kDa was found to be more effective (1.45 times) than the peptide fractions >3kDa however was less effective (1.19 times) than the whole hydrolysate regarding antioxidant activity. Similarly, ≤3kDa fraction of hydrolysate had 1.9 times higher ACE inhibition potential as compared to its whole hydrolysate. Conclusion: Overall, the study showed that the AGP employed whey protein hydrolysate has potential for use as a natural antioxidant and ACE inhibition agent.

Hydrolysates of mare's milk proteins. Immunochemical and physico-chemical characteristics

In recent years, the attention of specialists has been attracted by a significant increase in allergic diseases, especially among children. Enzymatic hydrolysis with the participation of peptidases is considered the most effective measure to reduce the allergenicity of milk proteins. The study of the features of protein proteolysis by various enzymes, aimed at establishing optimal process parameters, is the subject of research in the production of hydrolysates with specified properties: a certain peptide and amino acid profile, reduced antigenic properties. At the same time, for a correct preclinical assessment of the safety and potential effectiveness of specialized food products, it is not enough only to determine the molecular weight distribution of peptide fractions in the composition. A necessary additional and informative indicator of reducing the potential allergenicity of the hydrolysate is the degree of preservation of the antigenic properties of the initial protein (residual antigenicity), determined by immunochemical methods. The preparation and subsequent comprehensive assessment of enzymatic hydrolysates of mare's milk proteins is of considerable interest, since, according to the results of clinical studies, it was suggested that upon allergy to cow's milk proteins mare's milk can be considered hypoallergenic. The purpose of this work was to obtain enzymatic hydrolysates of mare's milk proteins using domestic enzymes and their further physico-chemical and immunochemical characteristics. The object of the study was mare's milk protein obtained from skimmed mare's milk powder as a protein substrate. Two samples of enzymes from various manufacturers of the Russian Federation were used in the work. The results of the conducted studies show that the peptide profiles of hydrolysates vary greatly depending on the enzyme used, however, the multiplicity of antigenicity reduction practically does not depend on the nature of the enzyme, and does not depend on an increase in the enzyme/substrate ratio. The peptide profile and the multiplicity of reducing the antigenicity of hydrolysates of mare's milk proteins are influenced by ultrafiltration treatment of the obtained hydrolysates, which allows reducing the content of antigenic structures in the final product by several orders of magnitude.

Characteristics and Bioactivities of Protein Hydrolysate from Cricket (Acheta domesticus) Powder Defatted Using Ethanol with Aid of Vacuum Impregnation.

Cricket is a potential proteinaceous source used for protein hydrolysate (PH) preparation, having several biological activities. Nevertheless, cricket has high lipid contents, which are susceptible to oxidation during PH preparation. Thus, ethanol was used together with vacuum impregnation (VI) to enhance defatting efficacy before PH preparation. Also, bioavailability of the digest of PH after gastrointestinal tract (GIT) digestion via the Caco-2 monolayer was assessed. Cricket powder was defatted using ethanol for 1-4 h. Lipid contents were decreased with enhancing time until 2 h. Additionally, the defatting efficacy was augmented when ethanol combined with VI at 4 cycles for 2 h (VI-E-2) was implemented. Lowered mono- and polyunsaturated fatty acid contents were also observed in the VI-E-2 sample. The VI-E-2 sample was used to prepare PH using Alcalase and Flavourzyme (0.2-0.4 units/g dry sample). PH prepared by Alcalase hydrolysis at 0.2 units/g dry sample (A-0.2) showed the higher ABTS radical-scavenging activity and FRAP, compared to that prepared by Flavourzyme hydrolysis (p < 0.05). Thus, the A-0.2 sample was selected for digestion via the GIT system. The obtained digest (500-1000 μg/mL) had bioavailability of peptides, depending on the levels used. Therefore, PH from defatted cricket powder could be a promising ingredient for food applications.

Characterization of the extracellular proteases from Bacillus inaquosorum strain E1-8 and its application in the preparation of hydrolysates from plant and animal proteins with antioxidant, antifreeze and anti-browning properties.

Bacillus inaquosorum strains is widely recognized for their plant-growth-promoting and biocontrol capabilities, yet their roles in protease production remain unclear. The present study aimed to comprehensively assess the protease-producing performance of B. inaquosorum strain E1-8, at the same time as exploring the novel application of agricultural Bacillus proteases in the preparation of protein hydrolysates for fresh-cut fruits preservation. First, genomic sequencing revealed the diversity of E1-8 proteases, indicating 15 putative extracellular proteases. Subsequently, the fermentation conditions for E1-8 protease production were optimized, with sweet potato powder and soybean meal identified as the most suitable carbon and nitrogen sources, respectively, resulting in a maximum protease activity of 321.48 U mL-1. Upon culturing the strain under these optimized conditions, only an S8 family serine protease and an M48 family metalloprotease were revealed by secretomic analysis and protease inhibitor assays. Additionally, the optimal protease conditions for generating protein hydrolysates from soy, pea, fish and porcine proteins were determined. The molecular weight of the hydrolysates primarily ranged from 2000 to 180 Da, with a total of 17 amino acids identified. The application of these hydrolysates demonstrated a 2,2-diphenyl-1-picrylhydrazyl (i.e. DPPH) scavenging activity ranging from 58.64% to 84.12%, significantly reducing of the melting peaks and the freezing points. Furthermore, the browning index of apple slices stored at 4 °C decreased by 14.81% to 22.15% on the second day, and similar effects were observed in fresh-cut banana stored at 4 °C for 7 days. The protein hydrolysates obtained exhibit remarkable antioxidant, antifreeze and anti-browning properties for fresh-cut fruits. © 2024 Society of Chemical Industry.

Effect of Chicken Feather Hydrolysate on Growth of Spinach through Soil Amendment Method: Unraveling A Potential Liquid Biofertilizer

The study aims to investigate the effectiveness of chicken feather hydrolysate for promoting the growth of Spinacia oleracea L., a commonly consumed leafy green vegetable. An earlier isolated and identified keratinolytic bacterial species Bacillus tropicus was utilized for the preparation of chicken feather hydrolysate through submerged fermentation. Minimal media which was supplemented with chicken feather was used for the preparation of hydrolysate. The bacterial strain degraded chicken feather within 4 days of incubation after which the feather hydrolysate was collected and tested to check plant growth promoting activity through the seed germination trials and greenhouse study. Upon characterization of feather hydrolysate, it was found that the hydrolysate was a cocktail of Nitrogen, Phosphorus and Potassium (NPK) as well as other micro elements needed for plant growth. Four different concentrations of feather hydrolysate were employed for both the seed germination and greenhouse study which ranged from 25% (v/v), 30% (v/v), 35% (v/v) and 40% (v/v) including a control group (CN) which was not supplemented with feather hydrolysate. The hydrolysate supplementation brought about plant growth in all the four test concentrations with 35% (v/v) giving the highest result of 14 cm and 27.6 mg/g for tested parameters like plumule length and total chlorophyll content, respectively. The same concentration supported maximum seed germination and highest radicle extension for the germination studies as well. This study investigates the efficacy of chicken feather hydrolysate in promoting spinach growth, elucidating its potential as a fertilizer.

The rolling-disc milling texture-sorting of raw straw facilitates the reduction of steam explosion intensity

Lignocellulosic fermentable sugars have been attracting increasing attention due to the developments of synthetic biology, yet the heterogeneity of the raw material results in high pretreatment intensities and the generation of numerous fermentation inhibitors. In this study, the rolling-disc milling texture-sorting was established for raw material to mitigate inhibitors generation at its source, specifically targeting the variances in mechanical strength among corn straw tissues. Through the texture-sorting of raw corn straw, the low modulus of elasticity tissues in corn straw were crushed into soft-texture sorts and the high-strength tissues were retained as hard-texture sorts. The results showed that the steam explosion pretreatment intensity required for soft-texture sorts decreased by 77.34 %, from 1125.96 (180 °C, 5 min) to 255.10 (160 °C, 5 min), compared to the hard-texture sorts. The concentrations of inhibitors in the enzymatic hydrolysate of soft-texture sorts decreased by 55.10 %, from 8.13 g/L/(100 g/L total sugars) to 3.65 g/L/(100 g/L total sugars), indicating an improved suitability for enzymatic hydrolysate preparation. Therefore, the rolling-disc milling texture-sorting method was proposed to provide a new approach for the preparation of lignocellulosic fermentable sugars.

Characterisation and comparison of enzymatically prepared donkey milk whey protein hydrolysates

This study evaluated the structural characteristics, processing properties, and antioxidant properties of hydrolysates prepared from donkey milk (DM) whey protein using different proteases (Alcalase, Neutrase, papain, and Flavourzyme). The results showed that enzymatic hydrolysis significantly increased hydrolysate solubility and reduced average particle size compared to those of DM whey protein. Neutrase and Flavourzyme hydrolysates exhibited higher degrees of hydrolysis (DH), along with elevated emulsification properties and surface hydrophobicity. The choice of protease influenced secondary and tertiary protein structures and amino acid composition. Enzymatic hydrolysis led to decreased molecular weight of DM whey proteins. Moreover, all hydrolysates exhibited higher fluorescence intensity at λmax compared to DM whey protein, implying distinct properties due to the varied impacts of the four proteases on DM whey protein structure. The preparation of hydrolysates from DM whey proteins using proteases contributes to the development of integrated-value DM products.

Food bioactive peptides: functionality beyond bitterness.

Bitter taste is an aversive taste because it is unconsciously associated with toxic compounds. However, a considerable variability in bitter sensitivity exists in those who have the genetic polymorphism for bitter taste receptors (TAS2Rs). Besides the oral cavity, TAS2Rs are present in many body tissues, including the gastrointestinal tract; therefore, they are crucial players both in the gustatory/hedonic system and in the homeostatic system, triggering numerous biological responses, including adipogenesis, carcinogenesis, or immunity. Bitter-tasting compounds are widely distributed in plant and animal foods and belong to many chemical classes. In this study, the evidence was reviewed on bitter peptides, considering the food sources, their formation in food under different processing and storage conditions and in the gastrointestinal tract during digestion, as well as their biological activities. Bitterness associated with peptides is due to the presence of hydrophobic amino acids in the C-terminus. The current literature mainly explores the enzymes and hydrolysis conditions, with the aim of reducing the formation of bitter peptides in hydrolysate preparation or food. Few studies highlight the bioactivity (namely, antihypertensive, antidiabetic, antioxidant, or immunity boosting), besides the bitterness. However, encapsulation of bitter peptides has been tentatively used to develop antihypertensive and antidiabetic supplements. In the era of personalized nutrition and precision medicine, the evidence available suggests the opportunity to use bitter bioactive peptides as functional ingredients in food. Such types of food may modulate a plethora of physiological mechanisms by targeting TAS2Rs in the gastrointestinal tract, thus modulating appetite sensations or gastrointestinal motility and discomfort according to individual nutritional needs and goals. More studies are needed to optimize the technological strategies to target TAS2Rs by bitter bioactive peptides, improve their stability in food, and validate the biological efficacy through well-designed in vivo studies.

Purification of Trypsin from Sardine (Sardina pilchardus) Viscera and Its Application in Preparation of Antioxidative Fish Protein Hydrolysates

Purification of Trypsin from Sardine (Sardina pilchardus) Viscera and Its Application in Preparation of Antioxidative Fish Protein Hydrolysates

A strategy to boost xanthine oxidase and angiotensin converting enzyme inhibitory activities of peptides via molecular docking and module substitution

Molecular docking and activity evaluation screened the dipeptide module GP with low xanthine oxidase (XOD) inhibitory activity and modules KE and KN with high activity, and identified them as low- and high-contribution modules, respectively. We hypothesized the substitution of low-contribution modules in peptides with high contributions would boost their XOD inhibitory activity. In the XOD inhibitory peptide GPAGPR, substitution of GP with both KE and KN led to enhanced affinity between the peptides and XOD. They also increased XOD inhibitory activity (26.4% and 10.3%) and decreased cellular uric acid concentrations (28.0% and 10.4%). RNA sequencing indicated that these improvements were attributable to the inhibition of uric acid biosynthesis. In addition, module substitution increased the angiotensin-converting enzyme inhibitory activity of GILRP and GAAGGAF by 84.8% and 76.5%. This study revealed that module substitution is a feasible strategy to boost peptide activity, and provided information for the optimization of hydrolysate preparation conditions.

Bioactive peptides as a novel strategy to prevent alcoholic liver injury.

Alcohol intake has become one of the leading risks to human health and wellness, among which acute and/or chronic alcohol-induced liver injury is a leading threaten, with few therapeutic options other than abstinence. In recent years, studies suggested that certain bioactive peptides from food sources could represent natural and safe alternatives for the prevention of alcoholic liver injury. Hence, this chapter focus on the advanced research on bioactive peptides exerting hepatoprotective activity against alcoholic liver injury. The main sources of protein, strategies for the preparation of hepatoprotective hydrolysates and peptides, underlying mechanisms of peptides on hepatoprotection, and possible structure-activity relationship between peptides and hepatoprotective activity were summarized and discussed, aiming to give a systematic insight into the research progress of hepatoprotective peptides. However, more efforts would be needed to give a clearer insight into the underlying mechanisms and structure-activity relationship before using hepatoprotective peptides as functional food ingredients or dietary supplements.

Tea (Camellia sinensis (L.) Kuntze) as an emerging source of protein and bioactive peptides: A narrative review.

Tea residues represent one of the major agricultural wastes that are generated after the processing of tea. They account for 21-28% of crude protein and are often discarded without the extraction of valuable proteins. Due to various bioactivity and functional properties, tea proteins are an excellent alternative to other plant-based proteins for usage as food supplements at a higher dosage. Moreover, their good gelation capacity is ideal for the manufacturing of dairy products, jellies, condensation protein, gelatin gel, bread, etc. The current study is the first to comprehend various tea protein extraction methods and their amino acid profile. The preparation of tea protein bioactive peptides and hydrolysates are summarized. Several functional properties (solubility, foaming capacity, emulsification, water/oil absorption capacity) and bioactivities (antioxidant, antihypertensive, antidiabetic) of tea proteins are emphasized.

Efficient preparation of oyster hydrolysate with aroma and umami coexistence derived from ultrasonic pretreatment assisted enzymatic hydrolysis

In the study, an efficient protease Neutrase®-Flavourzyme® (NF) was screened to prepare the umami-aroma flavor oyster hydrolysate. The effect of protease and ultrasonic pretreatment (UP) assisted by the optimal protease on the flavor substances was investigated. The results demonstrated that the optimal UP-NF (450 W) showed a higher amino acid nitrogen content of 0.34 g/100 mL compared to the NF, and 19 major aroma compounds including octanal, decanal, nonanal, benzaldehyde, 2-undecanone, and 1-octen-3-ol were obtained. Additionally, the free amino acid and fatty acid spectrum indicated that the formation of flavor compounds was primarily due to the oxidation of linoleic and linolenic acids and the degradation of amino acids. Furthermore, taste analysis proved that increased umami and saltiness resulted from the accelerated release of Glu, Asp and 5′-IMP. Overall, UP-NF proved to be an effective method for producing umami-aroma flavor, facilitating further processing of oyster products for the application.

The preparation, characterization and gastroprotective activity of fermented oyster hydrolysate

The preparation, characterization and gastroprotective activity of fermented oyster hydrolysate

Collagen hydrolysates from deer tendon: Preparation assisted with different ultrasound pretreatment times and promotion in MC3T3-E1 cell proliferation and antioxidant activities

In this study, the hydrolysates of deer tendons with antioxidant activity and promoting bone proliferation activity were prepared using ultrasound. The results indicated that ultrasound (60 min) treatment had noticeable effects on the structure and enzymolysis degree of collagen protein, including looser structure, less α-helix/β-turn, more hydrophobic groups, and a higher degree of hydrolysis in comparison with others. Besides, compared with unpretreated collagen hydrolysates (CPHs), the hydrolysates with ultrasound treatment (60 min) showed significant effects on bone proliferation (cell proliferation rate: 63.9 ± 4.8%), alkaline phosphatase activity (14.488 U/mg), and antioxidant activities (DPPH radical scavenging rate: IC50 =4.05 mg/mL and ferric ion reducing antioxidant power (FRAP): 0.17 mmol Fe2+/mg). CPHs (ultrasound 60 min) also could protect MC3T3-E1 cells from H2O2-induced damage by reducing the ROS and [Ca2+]i. Taken together, these results might provide a basis for the ultrasound-assisted preparation of potential antioxidant hydrolysates to be applied in the food industry.

Functional Properties and Proximate Analysis of Fish Waste Protein Hydrolysate Processed Using Enzymes

A huge production of fish and their processing waste give rise to by-products comprises up to 70% depending on the species, size and processing method. The waste includes visceral parts, head, frames, bones, skin and cut-offs are rich source of protein with high functional properties. It is generally discarded which is a wastage of nutrient source and leading to environmental issues. Therefore, it was aimed to utilized the by-products for maximum recovery of nutrients by enzyme hydrolysis method for the preparation of protein hydrolysate with the used of papain and pepsin for digestion following different hydrolysis conditions. With the hydrolysis of papain enzyme (1 to 6%), the protein content of finfish waste protein hydrolysate ranges from 19.17% ± 0.06 to 73.14% ± 0.08 and that of shellfish waste protein hydrolysate prepared with 5,10 and 15% papain enzyme showed 26.73% ± 0.04 to 40.4% ± 0.5 which is comparatively low. Whereas the highest protein content was observed in 1% pepsin enzyme treated finfish waste protein hydrolysate with 80.55% ± 0.07. Besides, the hydrolysates were composed of 6.91% ± 0.05 to 10.46% ± 0.05 (moisture content), 0.6% ± 0.01 to 2.4% ± 0.01 (ash content) and 0.02% ± 0.005 to 0.09% ± 0.005 (fat content). The hydrolysates were highly soluble ranges from 72.73% ± 0.05 to 93.83% ± 0.1 which indicates development of small size hydrophilic with highly solvated polypeptide particles. A reduced phenomena of foaming capacity and stability were observed in shellfish waste protein hydrolysate in contrast with finfish waste protein hydrolysate. Similar pattern was also resulted in emulsifying stability index. Whereas the emulsifying activity index was in the range of 6.15 ± 0.03 to 9.85 ± 0.07 m2/g. The water holding capacity of finfish and shellfish waste protein hydrolysate ranges from 3.4 to 4.23 gm/gm hydrolysate and 1.53 to 1.63 gm/gm hydrolysate respectively which is resulted by the difference in molecular weight of the peptide. The hydrolysates extracted from different sample with different enzyme and concentration at varying conditions were more or less similar ranges from 3.7 to 4.1 gm/gm protein hydrolysate (oil holding capacity). Hence, high protein content with good functional properties of the protein hydrolysate prepared with the utilization of fish waste is a positive impact on the attempt made to recover nutrient by enzymatic hydrolysis.

Enzymatic Preparation of Mushroom By-product Protein Hydrolysates (Mb-PPHs)

The excessive use of chemical fertilizers can cause severe environmental damage. In recent decades, the application of biostimulants to improve soil composition and stimulate plant growth has contributed significantly to environmental preservation. In this paper, we studied the production and characterization of an amino acid/peptide-enriched biostimulant using edible mushroom (Agaricus bisporus) by-products (tails and nonmarketable mushrooms) as raw materials and commercial proteases as hydrolytic agents. A single hydrolytic process using four different endoproteases, Alcalase®, L-450, Flavourzyme® or papain, and a sequential hydrolytic process using two proteases, an endoprotease and an exoprotease, Alcalase® + Flavourzyme® or L-450 + Flavourzyme), were conducted. A preevaluation of potential plant biostimulants was also carried out, testing the biostimulant capacity of single and sequential Mb-PPHs to stimulate maize seed germination and root growth, as well as the evaluation of the vigor index (VI), with very promising results.Graphical

Preparation of sunflower seed-derived umami protein hydrolysates and their synergistic effect with monosodium glutamate and disodium inosine-5'-monophosphate.

Sunflower seeds are rich in protein and can be an excellent raw material for the production of umami peptides. In this study, sunflower seed meal, which was defatted at a low temperature, was taken as the raw material, and proteins were separated, followed by hydrolyzation for 4h by flavourzyme® to obtain hydrolysates with strong umami intensity. These hydrolysates were deamidated using glutaminase to increase the umami intensity. The highest umami value of 11.48 was recorded for hydrolysates deamidated for 6h, and the umami intensity was determined. The umami hydrolysates mixed with 8.92mmol IMP + 8.02mmol MSG showed the highest umami value of 25.21. Different concentrations of ethanol were used for further separation of hydrolysates, and the highest umami value of 13.54 was observed for 20% ethanol fraction. The results of this study provide utilization method for sunflower seed meal protein and a theoretical basis for the preparation of umami peptides. PRACTICAL APPLICATION: A large number of sunflower seed meals after oil production are used as feed for livestock and poultry. Sunflower seed meal is rich in protein, and umami amino acid composition in sunflower seed meal is up to 25%-30%, which is potentially an excellent raw material for the production of umami peptides. The umami flavor and synergistic effect of obtained hydrolysates, with MSG and IMP, were analyzed in the present study. We intend to provide a novel way for utilization of protein from sunflower seed meal along with a theoretical basis for the preparation of umami peptides.