Influence Of Enzymatic Hydrolysis Research Articles

The Influence of Enzymatic Hydrolysis on Bee Pollen Antioxidant and Antibacterial Activities.

Bee pollen is one of the most valuable apitherapeutic products with high nutritional value. To obtain a higher diversity of compounds, higher bioactivity, and improve the release of nutrients from bee pollen, additional processing of the raw material may be applied: fermentation using microorganisms or hydrolysis using selective enzymes. This research aimed to determine the impact of enzymatic hydrolysis on the antioxidant and antibacterial activities of bee pollen. Bee pollen samples from Sweden, Spain, Netherlands, Italy, Poland, Denmark, Slovakia, Malta, and Lithuania were hydrolyzed using pure enzymes, including lipase, cellulase, protease, and amyloglucosidase, as well as enzyme mixtures such as Viscozyme® L and Clara-diastase. Total phenolic content, total flavonoid content, and antioxidant activity were analyzed spectrophotometrically. Antibacterial activity against Staphylococcus aureus, Listeria monocytogenes, Staphylococcus enteritidis, and Salmonella typhimurium was evaluated using the agar well diffusion assay. Obtained results revealed a positive effect of enzymatic hydrolysis on biologically active compound content and activity: total phenolic content increased by 1.1 to 2.5 times, total flavonoid content by 1.1 to 3.0 times, radical scavenging activity by 1.1-3.5 times, and antibacterial activity by 1.1 to 3.3 times. K-means clustering analysis grouped samples into 5-9 clusters and was dependent on the measured characteristic used as an input-total phenolic compounds content, total flavonoid content, antioxidant activity, and antibacterial activity against four different bacteria. Chemometrics showed, that the enzyme used for the hydrolysis had a higher impact on clustering results than the geographical origin of the samples.

Influence of Enzymatic Hydrolysis and Molecular Weight Fractionation on the Antioxidant and Lipase / α-Amylase Inhibitory Activities In Vitro of Watermelon Seed Protein Hydrolysates.

This study aims to evaluate the potential in vitro antioxidant and anti-obesity activities of watermelon seed protein hydrolysates (WSPH) obtained using different combinations of enzymes alcalase-proteinase K (ALC-PK) and alcalase-actinidin (ALC-ACT). There was a direct relationship between the degree of hydrolysis (DH) and the biological activities of the WSPH, with the highest DPPH (approximately 85%) and lipase inhibitory activities (≈59%) appreciated at DH of 36-37% and 33-35% when using ALC-PK and ALC-ACT, respectively. Following molecular weight fractionation, the ALC-PK WSPH < 3 kDa (F1) assayed at 1 mg.mL-1 had the highest DPPH-radical scavenging (89.22%), ferrous chelating (FC) (79.83%), reducing power (RP) (A 0.51), lipase inhibitory (71.36%), and α-amylase inhibitory (62.08%) activities. The amino acid analysis of ALC-PK WSPH and its fractions revealed a relationship between the biological activity of the extracts and their composition. High contents of hydrophobic amino acids, arginine, and aromatic amino acids were related to high antioxidant, lipase inhibitory, and α-amylase inhibitory activities in the extracts, respectively. Overall, this study revealed that underutilized protein sources such as WSPH, using the appropriate combination of enzymes, could result in the generation of new ingredients and compounds with powerful antioxidant and anti-obesity activities with promising applications as nutraceuticals or functional foods.

Wool Keratin Hydrolysates for Bioactive Additives Preparation.

The aim of this paper was to select keratin hydrolysate with bioactive properties by using the enzymatic hydrolysis of wool. Different proteolytic enzymes such as Protamex, Esperase, and Valkerase were used to break keratin molecules in light of bioactive additive preparation. The enzymatic keratin hydrolysates were assessed in terms of the physico-chemical characteristics related to the content of dry substance, total nitrogen, keratin, ash, cysteic sulphur, and cysteine. The influence of enzymatic hydrolysis on molecular weight and amino acid composition was determined by gel permeation chromatography (GPC) and gas chromatography-mass spectrometry (GC-MS) analyses. Antimicrobial activity of keratin hydrolysates was analysed against Fusarium spp., a pathogenic fungus that can decrease the quality of plants. The bioactivity of enzymatic hydrolysates was tested on maize plants and allowed us to select the keratin hydrolysates processed with the Esperase and Valkerase enzymes. The ratio of organised structures of hydrolysate peptides was analysed by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) deconvolution of the amide I band and may explain the difference in their bioactive behaviour. The most important modifications in the ATR spectra of maize leaves in correlation with the experimentally proven performance on maize development by plant length and chlorophyll index quantification were detailed. The potential of enzymatic hydrolysis to design additives with different bioactivity was shown in the case of plant growth stimulation.

Effect of enzymatic hydrolysis on the formation and structural properties of peanut protein gels

Abstract The limited enzymatic hydrolysis gelation method was investigated using peanut protein isolate (PPI) without any coagulators. A peanut protein gel could be formed by enzyme treatment with Alcalase at low temperature (50–70 °C). The influence of enzymatic hydrolysis on the rheological and physicochemical properties was investigated. Structural changes in the PPI were characterized by analyzing the subunits, chemical forces, surface hydrophobicity, fluorescence spectra, and circular dichroism (CD) spectra. The results revealed that enzymatic hydrolysis significantly affected the conarachin II protein of PPI, and had little influence on conarachin I and the basic subunits of arachin. Hydrophobic interaction was the main chemical force active in the peanut protein gel. An increase in the surface hydrophobicity coupled with red-shifts of the fluorescence spectra indicated that inner hydrophobic regions were exposed after hydrolysis, resulting in gel formation via hydrophobic interactions. The CD spectra showed that significant changes occurred in the secondary structure of PPI, where the ordered PPI structure formed a more open structure after enzymatic hydrolysis.

Influence of enzymatic hydrolysis on the properties of red rice starch

Starch is a technologically challenging polysaccharide that arouses the global interest in its modification to increase its applicability. Thus, the objective of the present work was to study the influence of enzymatic modification on the properties of red rice starch. The enzymatic modification was performed sequentially through the actions of α-amylase enzymes and amyloglucosidase using a design of experiments where several process conditions were used at sub-gelatinization temperatures. Then the native and modified starches were characterized for its physicochemical, physical, centesimal, phytochemical, morphological, and thermal properties. The enzymatic hydrolysis of the starch granules with α-amylase resulted in 4.46% hydrolysis and more 8.63% after the action amyloglucosidase. Results revealed that surface adsorption is more limiting than diffusion. The enzymatic hydrolysis process resulted in a reduction of the amylase and amylopectin content, as well as on all phytochemicals. After hydrolysis, granules starches presented surface alterations and tendency to become spherical. Small differences on crystallinity revealed that the enzymes had more influence on the amorphous fractions of the starch granules.

The Influence of Enzymatic Hydrolysis on Adsorption and Interfacial Dilatational Properties of Pumpkin (Cucurbita pepo) Seed Protein Isolate

Pumpkin seed protein isolate, PSPI, was enzymatically hydrolysed by alcalase to obtain pumpkin seed protein hydrolysate, PSPH. Kinetics of PSPI and PSPH adsorption layer formation at oil–protein solution interface and interfacial dilatational properties of the layers were investigated by the drop profile analysis tensiometer (PAT) in order to determine the influence of enzimatic hydrolysis on the interfacial properties of pumpkin seed proteins. The properties were investigated at different protein solution concentrations (0.0008–0.8 g/100 mL), ionic strengths (0–0.5 mol/L NaCl), and at two acidic pH (3 and 5, where PSPI’s pI = 5). It was found that both, PSPI and PSPH, contribute to an increase in the interfacial pressure, π, at the oil–protein solution interface and form the interfacial proteinaceous films. Dilatational elasticity, E’, of the interfacial films was found to be a few times higher than the dilatational viscosity, E”, regardless of the experimental conditions. The obtained diffusion rate and adsorption rate constants, kdiff and kads respectively, were higher for PSPH than for PSPI. kdiff was found to increase as protein concentration was increased, and to decrease as ionic strength was increased, for both PSPI and PSPH. At pI = 5, PSPH showed an increased π and E’, as well as mitigated influence of ionic strength on kads when compared to PSPI.

Influence of enzymatic hydrolysis on the allergenic reactivity of processed cashew and pistachio

Cashew and pistachio allergies are considered a serious health problem. Previous studies have shown that thermal processing, pressurization and enzymatic hydrolysis may reduce the allergenic properties of food by changing the protein structure. This study assesses the allergenic properties of cashew and pistachio after thermal treatment (boiling and autoclaving), with or without pressure (autoclaving), and multiple enzymatic treatments under sonication, by SDS-PAGE, western blot and ELISA, with serum IgE of allergic individuals, and mass spectroscopy. Autoclaving and enzymatic hydrolysis under sonication separately induced a measurable reduction in the IgE binding properties of pastes made from treated cashew and pistachio nuts. These treatments were more effective with pistachio allergens. However, heat combined with enzymatic digestion was necessary to markedly lower IgE binding to cashew allergens. The findings identify highly effective simultaneous processing conditions to reduce or even abolish the allergenic potency of cashew and pistachio.

Influence of enzymatic hydrolysis on solubility, interfacial and emulsifying properties of pumpkin (Cucurbita pepo) seed protein isolate

Pumpkin seed protein isolate (PSPI) was enzymatically hydrolyzed by alcalase and pepsin to obtain H1 and H2 hydrolysates, respectively. Influence of enzymatic hydrolysis on solubility, interfacial and emulsifying properties of PSPI was investigated as a function of pH (3–8) and ionic strength (0–1 mol/dm3 NaCl). Solubility of PSPI was lowest at pI = 5. Solubility of both hydrolysates was higher than solubility of PSPI, especially at pH close to pI. Increase in ionic strength brings about salting–in or salting–out effect in PSPI, H1, and H2 solutions, depending on pH of the solutions. Tensiometric investigation showed that PSPI, H1 and H2 adsorb at both air/protein solution and oil/protein solution interfaces regardless of pH and ionic strength. 20% emulsions of sunflower oil in a continuous phase consisting of 1 g/100 cm3 PSPI, H1 or H2 solutions at pH 3, 5 and 8, and ionic strengths of 0 and 0.5 mol/dm3 NaCl were prepared. Emulsifying performance of proteins depended on pH and ionic strength. PSPI failed to stabilize emulsions against droplet coalescence at pI = 5 regardless of ionic strength, and at pH 3 with 0.5 mol/dm3 NaCl, while both hydrolysates, H1 and H2, successfully stabilized emulsions no matter of pH and ionic strength. Smallest mean droplet diameter in emulsions of increased ionic strength (0.5 mol/dm3) were obtained when they were stabilized with H2, H1, and both PSPI and H1 at pH 3, 5, and 8, respectively. All emulsions were susceptible to creaming instability.

Influence of Enzymatic Hydrolysis on the Nutritional, Functional and Antioxidant Properties of Protein Hydrolysates Prepared from Seinat (Cucumis melo var. tibish) Seeds

The effect of enzymatic hydrolysis time on nutritional, functional properties and antioxidant activities of seinat seed protein isolates were studied. Hydrolysates of seinat seed protein isolates (HSSPIs) were prepared at different time (30-180 min), using two food-grade proteases. Trypsin was applied in the first hydrolysis stage followed by Pepsin. The hydrolysis time showed significant differences (p<0.05) on nutritional parameters such as essential amino acid score, essential amino acid index, biological value and predicted protein efficiency ratios. All the functional properties studied decreased as hydrolysis time increases. Antioxidant activities; (DPPH radical scavenging, ABTS radical scavenging and ferrous chelating) of HSSPIs increased with increase in hydrolysis time. Large amounts of small-sized peptides (3,000-5,000 Da) were observed with decreased as hydrolysis time increased (2.07-1.28%). The results in this study indicated that increase in hydrolysis time has positively affected nutritional and antioxidant properties of HSSPIs, but had a negative impact on the functional properties studied.

Influence of Enzymatic Hydrolysis on the Allergenicity of Roasted Peanut Protein Extract

Background: Peanut allergy is recognized as one of the most severe food allergies. Some studies have investigated the effects of enzymatic treatments on the in vitro immunological reactivity of members of the Leguminosae family, such as the soybean, chickpea and lentil. Nevertheless, there are only a few studies carried out with sera from patients with a well-documented allergy. Methods: Roasted peanut protein extract was hydrolyzed by the sequential and individual action of 2 food-grade enzymes, an endoprotease (Alcalase) and an exoprotease (Flavourzyme). Immunoreactivity to roasted peanut extract and hydrolyzed samples was evaluated by means of IgE immunoblot, ELISA and 2-dimensional electrophoresis using sera from 5 patients with a clinical allergy to peanuts and anti-Ara h 1, anti-Ara h 2 and anti-Ara h 3 immunoblots. Results: Immunoblot and ELISA assays showed an important decrease of IgE reactivity and Ara h 1, Ara h 2 and Ara h 3 levels in the first 30 min of hydrolyzation with Alcalase. In contrast, individual treatment with Flavourzyme caused an increase in IgE reactivity detected by ELISA at 30 min and led to a 65% inhibition of IgE reactivity at the end of the assay (300 min). Ara h 1 and the basic subunit of Ara h 3 were still recognized after treatment with Flavourzyme for 300 min. Conclusion: Hydrolysis with the endoprotease Alcalase decreases IgE reactivity in the soluble protein fraction of roasted peanut better than hydrolysis with the exoprotease Flavourzyme.

Influence of enzymatic hydrolysis on functionality of plant proteins

DSC was used to study the extent of denaturation of hemisphaericin and mexicain hydrolysates from corn gluten, soybean and sunflower meals. It was observed that the defatted meals studied exhibited only one broad peak transition. The data obtained demonstrated that the partial protein denaturation found with hemisphaericin or mexicain is correlated to modifications of functional properties. The two enzymes display different modes of action, according to the protein source.

Influence of Enzymatic Hydrolysis on Structure and Emulsifying Properties of Sardine (Sardina pilchardus) Protein Hydrolysates

ABSTRACTThe influence of enzymatic modification with Alcalase was studied on surface hydrophobicity, molecular weight distribution and emulsifying properties of ten hydrolysates from sardine differing in degree of hydrolysis. Surface hydrophobicity was evaluated fluorometrically using 1 anilino‐8‐naphthalenesulfonate (ANS); molecular weight distribution was determined by gel chromatography on a Superfine Sephadex G50. Emulsifying capacity and stability were evaluated as functions of pH values and product concentration, in comparison with sodium caseinate. Results showed emulsifying properties, surface hydrophobicity and the high molecular weight fraction decreased as degree of hydrolysis increased. Thus production of hydrolysates with desired molecular structures and emulsifying properties is possible.