In Vitro Multi-Bioactive Potential of Enzymatic Hydrolysis of a Non-Toxic Jatropha curcas Cake Protein Isolate.

The Jatropha curcas cake, a protein-rich by-product of biofuel production, was the subject of our study. We identified and quantified the ACE inhibitory, antioxidant, and antidiabetic activities of bioactive peptides from a Jatropha curcas L. var Sevangel protein isolate. The protein isolate (20.44% recovered dry matter, 38.75% protein content, and 34.98% protein yield) was subjected to two enzyme systems for hydrolysis: alcalase (PEJA) and flavourzyme (PEJF), recording every 2 h until 8 h had passed. The highest proteolytic capacity in PEJA was reached at 2 h (4041.38 ± 50.89), while in PEJF, it was reached at 6 h (3435.16 ± 59.31). Gel electrophoresis of the PEJA and PEJF samples showed bands corresponding to peptides smaller than 10 kDa in both systems studied. The highest values for the antioxidant capacity (DPPH) were obtained at 4 h for PEJA (56.17 ± 1.14), while they were obtained at 6 h for PEJF (26.64 ± 0.52). The highest values for the antihypertensive capacity were recorded at 6 h (86.46 ± 1.85) in PEJF. The highest antidiabetic capacity obtained for PEJA and PEJF was observed at 6 h, 68.86 ± 8.27 and 52.75 ± 2.23, respectively. This is the first report of their antidiabetic activity. Notably, alcalase hydrolysate outperformed flavourzyme hydrolysate and the cereals reported in other studies, confirming its better multi-bioactivity.

Influence of the Enzymatic Hydrolysis Using Flavourzyme Enzyme on Functional, Secondary Structure, and Antioxidant Characteristics of Protein Hydrolysates Produced from Bighead Carp (Hypophthalmichthys nobilis).

In the current study, bighead carp fish were used in conjunction with the flavourzyme enzyme to obtain (FPH) fish protein hydrolysates. The optimum conditions of the hydrolysis process included an enzyme/substrate ratio of 4% and a temperature of 50 °C and pH of 6.5. The hydrolysis time was studied and investigated at 1, 3, and 6 h, and the (DH) degree of hydrolysis was recorded at 16.56%, 22.23%, and 25.48%, respectively. The greatest yield value was 17.83% at DH 25.48%. By increasing the DH up to 25.48%, the crude protein and total amino acid composition of the hydrolysate were 88.19% and 86.03%, respectively. Moreover, more peptides with low molecular weight were formed during hydrolysis, which could enhance the functional properties of FPH, particularly the solubility property ranging from 85% to 97%. FTIR analysis revealed that enzymatic hydrolysis impacted the protein's secondary structure, as indicated by a remarkable wavelength of amide bands. Additionally, antioxidant activities were investigated and showed high activity of DDPH radical scavenging, and hydroxyl radical scavenging demonstrated remarkable activity. The current findings demonstrate that the functional, structural, and antioxidant characteristics of FPH might make it an excellent source of protein and suggest potential applications in the food industry.

Antioxidant potential and amino acid profile of ultrafiltration derived peptide fractions of spent hen meat protein hydrolysate.

Spent hen meat is considered as a category of waste generated by the poultry sector which can lead to serious environmental concerns if not disposed and utilized properly. In this work, spent hen meat was hydrolysed by 2% Flavourzyme (6.5 pH, 55 °C) followed by ultrafiltration to produce three peptide fractions with molecular weights > 10 kDa, 5-10 kDa and < 5 kDa. These fractions were evaluated for antioxidant potential, SDS PAGE and amino acid profile. The SDS PAGE profile demonstrated bands in the low molecular weight (< 10 kDa) region. Peptide fractions of < 5 kDa exhibited highest antioxidant activity and, essential as well as hydrophobic amino acid composition than whole hydrolysate and other peptide fractions. Incorporation of the identified hydrolysate fraction in food could improve its shelf stability while serving as a preventive component against human degenerative diseases.

The impact of Flavourzyme on flavor, antioxidant activity, and sensory attributes of salted grass carp by brine injection and brining.

BACKGROUND: Enzyme injection is vital for improving the sensory attributes and processing characteristics of meat products by enhancing proteolysis. However, studies regarding the appropriate dose addition for accelerating protein degradation in grass carp are minimal. This study aimed to investigate the impact of Flavourzyme® on the flavor quality and antioxidant activity of salted grass carp via brine injection and brining. RESULTS: Flavourzyme was added at doses of 0, 5, 10, 20, and 30 leucine aminopeptidase units (LAPU) per kilogram of raw meat. The results indicated that adding Flavourzyme promoted proteolysis, which was reflected by the enhanced total free amino acid content (from 3.7414 g kg-1 to 4.9160 g kg-1 in the brining group and from 3.8039 g kg-1 to 5.4061 g kg-1 in the injection group) and a decrease in salt soluble and insoluble protein (P < 0.05). The antioxidant activity was improved, and the thiobarbituric acid reactive substance value in salted carp decreased due to the higher content of the protein hydrolysis product (P < 0.05). All sensory attributes were improved significantly, especially when using brine injection (P < 0.05). Brine injection was helpful to diffuse the Flavourzyme, resulting in stronger proteolysis. CONCLUSION: The appropriate Flavourzyme dose was 10 LAPU kg-1 in the injection group and 20 LAPU kg-1 in the brining group. Therefore, moderate Flavourzyme addition was excellent in improving sensory attributes and storage characteristics, whereas injection represented a novel method to obtain a similar fish meat quality in a shorter time and with less added Flavourzyme. © 2021 Society of Chemical Industry.

Comparative functional and spectroscopic analysis of spent hen meat hydrolysate by individual and combined treatment of microbial proteases.

Simultaneous (Sm) and sequential (Sq) use of microbial proteases for the hydrolysis of spent hen/chicken meat from antioxidant potential perspective is relatively unexplored and requires attention. In this work, meat was hydrolyzed using Flavourzyme (Fz) and Alcalase (Ac), each at 1, 2, and 3% for 6 h as well as using both enzymes (at 2% each) in Sm and Sq treatment. Maximum attained %DPPH-RSA (Fz:68.25; Ac:77.18; Sm:59.82; and Sq:65.97) and FRAP (mM TEAC/g) values (Fz:3.77; Ac:2.56; Sm:2.54; and Sq:3.37) were measured as a function of hydrolysis time. The highest (23.38%) and lowest (10.68%) degree of hydrolysis (DH) was obtained with 3% Ac and 1% Fz, respectively. FTIR spectroscopy clearly revealed changes in the secondary structure of proteins. SDS PAGE profiling of hydrolysates showed that Fz produces low molecular weight peptides (2-75 kDa) as compared to Ac or its combination with Ac. As per the results of this study, Sq enzyme treatment is recommended for preparing spent hen meat hydrolysate with higher functional attributes for possible use as functional food/nutraceutical.

Effect of proteases and alcohols used for debittering on characteristics and antioxidative activity of protein hydrolysate from salmon frames.

Protein hydrolysates were obtained from salmon frame using Alcalase or Flavourzyme at 3% (w/w protein) for 180 min. Protein hydrolysates prepared using Alcalase (HA) and Flavourzyme (HF) had DH and yield of 25.1-26.9% and 28.5-32.3 g/100 g sample, respectively. HF showed lower bitterness score (5.78) than that of HA (8.68) (P < 0.05). When HA and HF were further subjected to debittering with 2-butanol or isopropanol, the recovery of 77.88-81.60% was obtained (P < 0.05). HF and HA debittered with 2-butanol possessed less bitterness score, 3.60 and 3.77, respectively (P < 0.05). Surface hydrophobicity of 81.4 and 124.8 was attained when HF and HA were debittered with 2-butanol (P < 0.05). Selected debittered hydrolysates, produced using Flavourzyme, followed by fractionation using 2-butanol (HF-B) contained glutamic acid/glutamine (15.14 g/100 g), aspartic acid/asparagine (10.07 g/100 g) and glycine (9.30 g/100 g) as the predominant amino acids. HF-B had the decreased ABTS radical scavenging activity and metal chelating activity. A280 of peptides separated by gel filtration was lowered to some extent and coincided with the lower bitterness score and surface hydrophobicity. Thus, debittered protein hydrolysate from salmon frame could serve as a nutritive ingredient at high levels in health promoting foods.

Impact of sequential enzymatic hydrolysis on antioxidant activity and peptide profile of casein hydrolysate.

ABSTRACT: This paper shows the potential of dual enzyme approach on antioxidant activity of casein hydrolysates. Casein was hydrolysed using the proteolytic enzymes alcalase, flavourzyme in isolation and in sequential order. Casein hydrolysates were evaluated for the degree of hydrolysis, antioxidant activity, molecular weight distribution patterns and peptide sequence. Casein hydrolysate produced by the sequential hydrolysis of alcalase and flavourzyme showed higher degree of hydrolysis and antioxidant activity as compared to hydrolysate obtained by individual enzymes. In size exclusion chromatograph of casein hydrolysate S3, peptides with molecular weight of 0.57 kDa share 12% area in total area of chromatogram which was 10 times higher than that of hydrolysate S1 and nearly half of that of hydrolysate S2. On subjecting to HPLC-TOF-ESI separation potential antioxidant peptides were identified. The peptide sequence VLPVPQ along with potential fragments was identified in hydrolysate S1 and S2 and HPHPHLS along with its potential sequence was identified in hydrolysate S1, S2 and S3. Sequential hydrolysis of casein showed better antioxidant activity and peptide profile in less duration as compared to the casein hydrolysate obtained by individual enzyme.

Sequence Identification of Bioactive Peptides from Amaranth Seed Proteins (Amaranthus hypochondriacus spp.).

Amaranthus hypochondriacus spp. is a commonly grown cereal in Latin America, known for its high protein content. The objective of this study was to separate and identify bioactive peptides found in amaranth seeds through enzymatically-assisted hydrolysis using alcalase and flavourzyme. Hydrolysis was carried out for each enzyme separately and compared to two-step continuous process where both enzymes were combined. The biological activity of the resulting three hydrolysates was analyzed, finding, in general, higher bioactive potential of the hydrolysate obtained in a continuous process (combined enzymes). Its fractions were separated by RP-HPLC, and their bioactivity was analyzed. In particular, two fractions showed the highest biological activity as ACE inhibitors with IC50 at 0.158 and 0.134, thrombin inhibitors with IC50 of 167 and 155, and antioxidants in ABTS assay with SC50 at 1.375 and 0.992 mg/L, respectively. Further sequence analysis of the bioactive peptides was carried out using MALDI-TOF, which identified amino acid chains that have not been reported as bioactive so far. Bibliographic survey allowed identification of similarities between peptides reported in amaranth and other proteins. In conclusion, amaranth proteins are a potential source of peptides with multifunctional activity.

Characterization and Antioxidant and Angiotensin I-Converting Enzyme (ACE)-Inhibitory Activities of Gelatin Hydrolysates Prepared from Extrusion-Pretreated Milkfish (Chanos chanos) Scale.

Fish gelatin hydrolysates have been shown to possess various biological activities due to their unique Gly-Pro-Y and Gly-X-Hyp sequences. In the current study, fish gelatin was extracted from non-extruded milkfish scale (FSG1) or extrusion-pretreated milkfish scale (FSG2); extracted gelatins were hydrolyzed with different combinations of Flavourzyme and Alcalase to give four different hydrolysates, namely: FSGH1 (FSG1 hydrolyzed with Flavourzyme), FSGH2 (FSG1 hydrolyzed with Alcalase + Flavourzyme), FSGH3 (FSG2 hydrolyzed with Flavourzyme), and FSGH4 (FSG2 hydrolyzed with Alcalase + Flavourzyme). The extrusion-pretreatment process enhanced the extraction yield of gelatin from fish scale. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared (FTIR) analyses showed the extracts FSG1 and FSG2 possessed characteristics of gelatin. Moreover, the physicochemical characteristics of FSGH1⁻FSGH4 were examined by analyses of their degree of hydrolysis, amino acid composition, UV spectrum, FTIR spectrum, molecular weight, and RP-HPLC profile. Additional biological functional analyses showed that all of the studied gelatin hydrolysates FSGH1⁻FSGH4 possessed antioxidant activity dose-dependently as revealed by DPPH scavenging, ABTS scavenging, and reducing power analyses. In addition, FSGH2 and FSGH4 showed higher angiotensin-I-converting enzyme (ACE)-inhibitory activity as compared to FSGH1 and FSGH3. Taken together, FSGH2 and FSGH4 showed high antioxidant activity and potent anti-ACE activity. Due to the potential antioxidant and antihypertensive properties of FSGH2 and FSGH4, further research is needed to explore their possible use as natural supplementary raw materials in food and nutraceutical products.

Antioxidant activity of whey protein hydrolysates in milk beverage system.

The aim of the present study was to evaluate the antioxidant activity of flavoured milk enriched with antioxidative whey protein hydrolysates (WPHs) by radical scavenging method. Whey protein concentrate (WPC) was hydrolyzed by using three commercial proteases; flavouzyme, alcalase and corolase PP and these WPHs were analyzed for degree of hydrolysis and antioxidant activity. The antioxidant activities of these WPHs were evaluated using ABTS method. Trolox equivalent antioxidant activity of all the hydrolysates i.e. flavourzyme (0.81 ± 0.04), alcalase (1.16 ± 0.05) and corolase (1.42 ± 0.12) was higher than the WPC (0.19 ± 0.01). Among these, whey protein hydrolysates prepared using corolase showed maximum antioxidant activity. Total 15 ß-lactoglobulin, 1 α-lactoalbumin, and 6 ß-casein derived peptide fragments were identified in the WPHs by LC-MS/MS. Due to their size and characteristic amino acid composition, all the identified peptides may contribute for the antioxidant activity. The strawberry and chocolate flavoured milk was supplemented with WPC and WPHs and 2 % addition has shown increase in antioxidant activity upto 42 %. The result suggests that WPH could be used as natural biofunctional ingredients in enhancing antioxidant properties of food products.

The encapsulation of flavourzyme in nanoliposome by heating method.

The main objective of this study was to use heating method (HM) to prepare liposome without employing any chemical solvent or detergent. Plackett-Burman design (PBD) was applied for the screening of significant process variables including the lecithin proportion, the cholesterol/lecithin ratio, the pH of solution for liposome preparation, the enzyme/lecithin ratio, the stirring time, the process temperature, the speed of stirrer, the ratio of stirrer to the tank diameter, the application of homogenization, the method of adding enzyme and centrifugation conditions on the encapsulation efficiency (EE %) of liposome and the activity of liposomal Flavourzyme (LAPU(-1)) (P < 0.05). Then, the response surface methodology based on the central composite design (CCD) was applied for the evaluation of the impacts of the significant mentioned variables on the EE (%) and the activity of the liposomal Flavourzyme. The results indicated that the lecithin proportion and the stirring time were the major influential variables for both responses. The most suitable formulation of the Flavourzyme-loaded liposome is 4.5 % lecithin, 45 °C temperature, 5 % Flavourzyme/lecithin ratio, 30 min stirring time and medium pH of 6. Under suitable operating conditions, the EE of liposome and the activity of the liposomal Flavourzyme were achieved as 26.5 % and 9.96 LAPU ml(-1), respectively. AFM technique and size distribution clearly showed the diameter of 189 nm for the spherical shape of the Flavourzyme- loaded nanoliposome.

Rheological properties and microstructure of wheat flour dough systems with enzyme-hydrolyzed mashed potatoes.

The interest in incorporating potatoes into wheat dough is increasing. However, potatoes exhibit significant viscosity during thermal processing, affecting product processing and quality. This study aims to find an effective method to reduce the viscosity of mashed potatoes. We aimed to compare the effects of different enzymes (α-amylase, ß-amylase, and flavourzyme) and concentrations (0.01%, 0.05%, and 0.1%) on the micromorphology and rheological properties of mashed potatoes and potato-wheat dough. The impact of flavourzyme was the most significant (p<0.05). When enzyme concentration increased, viscosity decreased, and the degree of structural damage, indicated by increased porosity. Notably, the addition of flavourzyme can increase the content of sweet and umami free amino acids, improving the flavor of mashed potatoes. The scanning electron microscopy and confocal laser scanning microscopy images of potato-wheat dough revealed that enzyme-hydrolyzed mashed potatoes had improved homogeneity, reestablished the dough continuity, and strengthened the three-dimensional structure comprising proteins and starch. Notably, flavourzyme demonstrated the most significant effect on enhancing the protein-starch network structure. This was attributed to the exposure of functional groups resulting from protein hydrolysis, facilitating interaction with starch molecules. Our findings indicate that the addition of 0.1% flavourzyme (500 LAPU/g, pH 5.5, 55 ± 2°C, 30 min treated) was the most effective in reducing viscosity and reconstructing the gluten network. Enzymatic hydrolysis plays a vital role in the production of high-quality potato products, with particular importance in the baking industry, where flavourzyme exhibits significant potential. PRACTICAL APPLICATION: Enzymatic hydrolysis plays a vital role in the production of high-quality potato products, with particular importance in the baking industry, where flavourzyme exhibits significant potential.

Enzymatic hydrolysis improves the encapsulation properties of rice bran protein by increasing retention of anthocyanins in microparticles of grape juice.

There is a growing demand for the food industry to find appealing matrices that display a clean and sustainable label capable of replacing animal proteins in the encapsulation market for natural pigments. Therefore, this study evaluated the impact of enzymatic hydrolysis by Flavourzyme protease on the encapsulation properties of rice bran proteins, aiming to protect anthocyanins in grape juice microparticles. To achieve this, rice bran protein hydrolysates (RPH) with low (5%, LRPH), medium (10%, MRPH), and high (15%, HRPH) degrees of hydrolysis (DH) were used combined with maltodextrin as carrier agents for the microencapsulation of grape juice by spray drying. The feed solutions contained 1 g of carrier agents (CA)/g of soluble solids from the juice (SS) and protein: a 15% CA ratio. Non-hydrolyzed rice protein was used as a carrier agent to obtain a control sample to evaluate the effect of enzymatic hydrolysis on the microencapsulation of grape juice. Protein modification increased the surface activity of the protein and its ability to migrate to the surface of the microparticles, forming a protective film, as observed by X-ray photoelectron spectroscopy. Using HRPH as a carrier agent combined with maltodextrin improved the internal and total anthocyanin retention, antioxidant capacity measured by DPPH and ABTS+ assays, and powder recovery compared to the control sample, and increased DH reduced particle size and powder stickiness. These particles were more homogeneous, rough, and without cracks. The microencapsulation efficiency was above 70%. All powders exhibited low values of hygroscopicity and degree of caking. Therefore, enzymatic hydrolysis proves to be a promising alternative for improving rice bran protein's encapsulating properties since using RPH as an encapsulating agent conferred greater protection of anthocyanins in microparticles. Moreover, the HRPH sample exhibited the most favorable outcomes overall, indicating its potential for prospective utilization in the market, supported by its elevated Tg.

Synergistic Effects of Combined Flavourzyme and Floating Electrode-Dielectric Barrier Discharge Plasma on Reduction of Escherichia coli Biofilms in Squid (Todarodes pacificus).

This study investigated the synergistic effect of combining flavourzyme, a natural enzyme, and floating electrode-dielectric barrier discharge (FE-DBD) plasma (1.1 kV, 43 kHz, N2 1.5 m/s) treatment, a non-thermal decontamination technology, against Escherichia coli biofilms in squid. E. coli (ATCC 35150 and ATCC 14301) biofilms were formed on the surface of squid and treated with different minimum inhibitory concentrations (MICs) of flavourzyme (1/8; 31.25 µL/mL, 1/4; 62.5 µL/mL, 2/4; 125 µL/mL, and 3/4 MIC; 250 µL/mL) and FE-DBD plasma (5, 10, 30, and 60 min). Independently, flavourzyme and FE-DBD plasma treatment decreased by 0.26-1.71 and 0.19-1.03 log CFU/cm2, respectively. The most effective synergistic combination against E. coli biofilms was observed at 3/4 MIC flavourzyme + 60 min FE-DBD plasma exposure, resulting in a reduction of 1.55 log CFU/cm2. Furthermore, the combined treatment exhibited higher efficacy in E. coli biofilm inactivation in squid compared to individual treatments. The pH values of the synergistic combinations were not significantly different from those of the untreated samples. The outcomes indicate that the combined treatment with flavourzyme and FE-DBD plasma can effectively provide effective control of E. coli biofilms without causing pH changes in squid. Therefore, our study suggests a new microbial control method for microbial safety in the seafood industry.

Investigation of the peptides with calcium chelating capacity in hydrolysate derived from spent hen meat.

Calcium peptide chelates are developed as efficient supplements for preventing calcium deficiency. Spent hen meat (SHM) contains a high percentage of proteins but is generally wasted due to the disadvantages such as hard texture. We chose the underutilized SHM to produce peptides to bind calcium by proteolysis and aimed to investigate chelation between calcium and peptides in hydrolysate for a sustainable purpose. The optimized proteolysis conditions calculated from the result of response surface methodology for two-step hydrolysis were 0.30% (wenzyme/wmeat) for papain with a hydrolysis time of 3.5 h and 0.18% (wenzyme/wmeat) for flavourzyme with a hydrolysis time of 2.8 h. The enzymatic hydrolysate (EH) showed a binding capacity of 63.8 ± 1.8 mg calcium/g protein. Ethanol separation for EH improved the capacity up to a higher value of 68.6 ± 0.6 mg calcium/g protein with a high association constant of 420 M-1 (25°C) indicating high stability. The separated fraction with a higher amount of Glu, Asp, Lys, and Arg had higher calcium-binding capacity, which was related to the number of ─COOH and ─NH2 groups in peptide side chains according to the result from amino acid analysis and Fourier transform infrared spectroscopy. Two-step enzymatic hydrolysis and ethanol separation were an efficient combination to produce peptide mixtures derived from SHM with high calcium-binding capacity. The high percentage of hydrophilic amino acids in the separated fraction was concluded to increase calcium-binding capacity. This work provides foundations for increasing spent hen utilization and developing calcium peptide chelates based on underutilized meat.

Hydrolysis of chickpea proteins with Flavourzyme immobilized on glyoxyl-agarose gels improves functional properties.

Chickpea protein isolate was hydrolyzed using Flavourzyme immobilized on glyoxyl-agarose beads by multipoint covalent attachment. This Flavourzyme-glyoxyl derivative, produced after 1 h of immobilization at 4 °C followed by 5.5 h at room temperature, presented approximately 51% of the endoprotease activity of Flavourzyme but was around 700 times more stable than soluble enzyme. Chickpea protein hydrolysates ranging from 1% to 10% degree of hydrolysis were produced and their chemical composition was very close to that of protein isolate used as starting material. Solubility, oil absorption, emulsifying activity and stability, and foaming capacity and stability were determined. All protein hydrolysates showed higher solubility than intact proteins, especially at pHs near isoelectric point of native chickpea proteins. Moreover, all hydrolysates had better functional properties, except emulsifying activity, than the original protein isolate.

Enzymatic Hydrolysis of Tenebrio molitor (Mealworm) Using Nuruk Extract Concentrate and an Evaluation of Its Nutritional, Functional, and Sensory Properties.

Enzymatic protein hydrolysis is a well-established method for improving the quality of dietary proteins, including edible insects. Finding effective enzymes from natural sources is becoming increasingly important. This study used nuruk extract concentrate (NEC), an enzyme-rich fermentation starter, to produce protein hydrolysate from defatted Tenebrio molitor (also called mealworm, MW). The nutritional, functional, and sensorial properties of the hydrolysate were then compared to those obtained using commercial proteases (alcalase and flavourzyme). The protease activities of the crude nuruk extract (CNE), NEC, alcalase, and flavourzyme were 6.78, 12.71, 11.07, and 12.45 units/mL, respectively. The degree of hydrolysis and yield of MW hydrolysis by NEC were 15.10 and 35.92% (w/w), respectively. MW hydrolysate was obtained using NEC and had a significantly higher free amino acid content (90.37 mg/g) than alcalase (53.01 mg/g) and flavourzyme (79.64 mg/g) hydrolysates. Furthermore, the NEC hydrolysis of MW increased the antioxidant and angiotensin-converting enzyme inhibitory activity, with IC50 values of 3.07 and 0.15 mg/mL, respectively. The enzymatic hydrolysis also improved sensory properties, including umaminess, sweetness, and saltiness. Overall, this study found that the NEC hydrolysis of MW outperformed commercial proteases regarding nutritional quality, sensory attributes, and biological activity. Therefore, nuruk could potentially replace commercial proteases, lowering the cost of enzymatic protein hydrolysis.

Novel antihypertensive peptides from lupin protein hydrolysate: An in-silico identification and molecular docking studies.

Application of non-thermal treatment to proteins prior to enzymatic hydrolysis can facilitate the release of novel bioactive peptides (BPs) with unique biological activities. In this study, lupin protein isolate was pre-treated with ultrasound and hydrolysed using alcalase and flavourzyme to produce alcalase hydrolysate (ACT) and flavourzyme hydrolysate(FCT). These hydrolysates were fractionated into 1, 5, and 10 kDa molecular weight fractions using a membrane ultrafiltration technique. The in vitro angiotensin-converting enzyme (ACE) studies revealed that unfractionated ACT (IC50 = 3.21 mg mL-1) and FCT (IC50 = 3.32 mg mL-1) were more active inhibitors of ACE in comparison to their ultrafiltrated fractions with IC50 values ranging from 6.09 to 7.45 mg mL-1. Molecular docking analysis predicted three unique peptides from ACT (AIPPGIPY, SVPGCT, and QGAGG) and FCT (AIPINNPGKL, SGNQGP, and PPGIP) as potential ACE inhibitors. Thus, unique BPs with ACE inhibitory effects might be generated from ultrasonicated lupin protein.

The effect of hydrolyzed sesame meal protein on the quality and shelf life of hamburgers during refrigerated storage.

In this study, to improve the quality and shelf life of hamburgers, sesame meal protein hydrolysates (SPH) were produced using two enzymes of alcalase and flavourzyme and then four hamburger treatments: T1: control (10% soybean), T2: 1% SPH + soybean 9%, T3: 2% SPH + soybean 8%, and T4: 3% SPH + soybean 7% were prepared. Physicochemical properties were analyzed at the beginning of the storage period; microbial and chemical quality was evaluated at intervals of 0, 4, 8, 12, and 16 days. The results of SPH showed that alcalase enzyme can produce a SPH with a higher antioxidant properties (DPPH, FRAP, and beta-carotene-linoleic acid) (P < 0.05); therefore, this SPH was used for hamburger properties. According to the results, with the addition of SPH, moisture, fat, texture firmness decreased, protein, and brightness increased (P < 0.05), and all treatments had the allowable range. SPH replacement with soybean slowed down the increasing trend of oxidation and microbial spoilage (P < 0.05). In general, better results were observed in T3 and T4, which had a permissible range chemical and microbial index until the end of the storage period, as well as these treatments inhibited the growth of Staphylococcus aureus and Escherichia coli. Only T3 was approved by the evaluators.

Microencapsulation of a Commercial Food-Grade Protease by Spray Drying in Cross-Linked Chitosan Particles.

In this study, the use of spray-drying technology for encapsulating Flavourzyme® (protease-peptidase complex) was evaluated to overcome the limitations (low encapsulation efficiency and no large-scale production) of other encapsulation processes. To the best of our knowledge, spray drying has not been applied previously for the immobilization of this enzyme. Firstly, bovine serum albumin (BSA), as a model protein, was encapsulated by spray drying in chitosan and tripolyphoshate (TPP) cross-linked-chitosan shell matrices. The results showed that the chitosan-TPP microcapsules provided a high encapsulation efficiency and better protein stability compared to the non-crosslinked chitosan microcapsules. The effect of enzyme concentration and drying temperature were tested during the spray drying of Flavourzyme®. In this regard, an activity yield of 88.0% and encapsulation efficiency of 78.6% were obtained with a concentration of 0.1% (v/v) and an inlet temperature of 130 °C. Flavourzyme®-loaded chitosan microcapsules were also characterized in terms of their size and morphology using scanning electron microscopy and laser diffractometry.