Production and characterization of anti-hypertensive and anti-diabetic peptides from fermented sheep milk with anti-inflammatory activity: in vitro and molecular docking studies.

BACKGROUND: The present study aimed to evaluate the anti-hypertensive and anti-diabetic activities from biologically active peptides produced by fermented sheep milk with Lacticaseibacillus paracasei M11 (MG027695), as well as to purify and characterize the angiotensin-converting enzyme (ACE) inhibitory and anti-diabetic peptides produced from fermented sheep milk. RESULTS: After 48 h of fermentation at 37 °C, sheep milk demonstrated significant changes in anti-diabetic effects and ACE-I effects, with inhibition percentages observed for ACE inhibition (76.32%), α-amylase (70.13%), α-glucosidase (70.11%) and lipase inhibition (68.22%). The highest level of peptides (9.77 mg mL-1) was produced by optimizing the growth conditions, which included an inoculation rate of 2.5% and a 48 h of incubation period. The comparison of molecular weight distributions among protein fractions was conducted through sodium dodecyl-sulfate polyacrylamide gel electrophoresis analysis, whereas spots were separated using 2D gel electrophoresis according to both the molecular weight and pH. Peptide characterization with ultra-filtration membranes at 3 and 10 kDa allowed the study to assess molecular weight-based separation. Nitric oxide generated by lipopolysaccharide and the secretion of pro-inflammatory cytokines in RAW 264.7 immune cells were both inhibited by sheep milk fermented with M11. Fourier-transform infrared spectroscopy was employed to assess changes in functional groups after fermentation, providing insights into the structural changes occurring during fermentation. CONCLUSION: The present study demonstrates that fermentation with L. paracasei (M11) led to significant changes in fermented sheep milk, enhancing its bioactive properties, notably in terms of ACE inhibition and anti-diabetic activities, and the generation of peptides with bioactive properties has potential health benefits. © 2024 Society of Chemical Industry.

Peptidomics-based identification of antihypertensive and antidiabetic peptides from sheep milk fermented using Limosilactobacillus fermentum KGL4 MTCC 25515 with anti-inflammatory activity: in silico, in vitro, and molecular docking studies.

This study investigated the synthesis of bioactive peptides from sheep milk through fermentation with Limosilactobacillus fermentum KGL4 MTCC 25515 strain and assessed lipase inhibition, ACE inhibition, α-glucosidase inhibition, and α-amylase inhibition activities during the fermentation process. The study observed the highest activities, reaching 74.82%, 70.02%, 72.19%, and 67.08% (lipase inhibition, ACE inhibition, α-glucosidase inhibition, and α-amylase inhibition) after 48 h at 37°C, respectively. Growth optimization experiments revealed that a 2.5% inoculation rate after 48 h of fermentation time resulted in the highest proteolytic activity at 9.88 mg/mL. Additionally, fractions with less than 3 kDa of molecular weight exhibited superior ACE-inhibition and anti-diabetic activities compared to other fractions. Fermentation of sheep milk with KGL4 led to a significant reduction in the excessive production of NO, TNF-α, IL-6, and IL-1ß produced in RAW 267.4 cells upon treatment with LPS. Peptides were purified utilizing SDS-PAGE and electrophoresis on 2D gels, identifying a maximum number of proteins bands ranging 10-70 kDa. Peptide sequences were cross-referenced with AHTPDB and BIOPEP databases, confirming potential antihypertensive and antidiabetic properties. Notably, the peptide (GPFPILV) exhibited the highest HPEPDOCK score against both α-amylase and ACE.

Fabrication of jamun seed starch/tamarind kernel xyloglucan bio-nanocomposite films incorporated with chitosan nanoparticles and their application on sapota (Manilkara zapota) fruits.

The present work develops bio-nanocomposite packaging films by valorizing agricultural byproducts jamun seed starch (JaSS) and tamarind kernel xyloglucan (XG), and adding varying concentrations of chitosan nanoparticles (ChNPs). The blending of JaSS and XG promotes a dense polymer network in the composite films with enhanced packaging attributes. However, ChNPs incorporation significantly reduced the viscosity and dynamic moduli of the JaSS/XG film-forming solutions. The FTIR and XRD results reveal improved intermolecular interactions and crystallinity. The DSC and TGA thermograms showed improved thermal stability in the ChNP-loaded JaSS/XG films. The addition of 3 % w/w ChNPs significantly enhanced the tensile strength (20.42 MPa), elastic modulus (0.8 GPa), and contact angle (89°), along with reduced water vapor transmission rate (13.26 g/h.m2) of the JaSS/XG films. The films exhibited strong antimicrobial activity against Bacillus cereus and Escherichia coli. More interestingly, the JaSS/XG/ChNPs coating on the sapota fruits retarded the weight loss and color change up to 12 days of storage. Overall, the JaSS/XG/ChNP bio-nanocomposites are promising packaging materials.

Variations in Microstructural and Physicochemical Properties of Soy Wax/Soybean Oil-Derived Oleogels Using Soy Lecithin.

Emerging natural-based polymers and materials progress and new technology innovations open the way for unique food products with high nutritional value development. In this regard, oleogel may be essential in replacing fatty acids from food products. In this study, we researched the effects of varied soy lecithin (SYL) concentrations on the various physicochemical characteristics of soy wax (SW)/refined soybean oil (RSO) oleogels. These oleogels had a soft texture. The microscopic analysis of the oleogels suggested that the thickness, length, and density of the wax crystals (needle-shaped) varied as the SYL content was changed. Colorimetric analysis indicated that the oleogels were slightly yellowish. FTIR spectrometry helped analyze the functional groups of the raw materials and the oleogels. All the functional groups present in the raw materials could be accounted for within the oleogels. The only exception is the hydrogen-bonding peak in SW, which was not seen in the FTIR spectrum of the oleogels. It was found that at a critical SYL content, the oleogel showed a stable and repeatable wax network structure. This can be described by the presence of the uniformly distributed fat crystal network in the sample. The DSC analysis revealed that the oleogel samples were thermo-reversible, with their melting and crystallization temperatures ~43 °C and ~22 °C, respectively. In gist, it can be concluded that the incorporation of SYL can impact the color, wax crystal network characteristics, thermal characteristics, and mechanical characteristics of the oleogels in a composition-dependent manner.

Variations in Microstructural and Physicochemical Properties of Candelilla Wax/Rice Bran Oil-Derived Oleogels Using Sunflower Lecithin and Soya Lecithin.

Candelilla wax (CW) is a well-known oleogelator that displays tremendous oil-structuring potential. Lecithin acts as a crystal modifier due to its potential to alter the shape and size of the fat crystals by interacting with the wax molecules. The proposed work is an attempt to understand the impact of differently sourced lecithin, such as sunflower lecithin (SFL) and soya lecithin (SYL), on the various physicochemical properties of CW and rice bran oil (RBO) oleogels. The yellowish-white appearance of all samples and other effects of lecithin on the appearance of oleogels were initially quantified by using CIELab color parameters. The microstructural visualization confirmed grainy and globular fat structures of varied size, density, packing, and brightness. Samples made by using 5 mg of SFL (Sf5) and 1 mg of SYL (Sy1) in 20 g showed bright micrographs consisting of fat structures with better packing that might have been due to the improvised crystallinity in the said samples. The FTIR spectra of the prepared samples displayed no significant differences in the molecular interactions among the samples. Additionally, the slow crystallization kinetics of Sf5 and Sy1 correlated with better crystal packing and fewer crystal defects. The DSC endotherm displayed two peaks for melting corresponding to the melting of different molecular components of CW. However, all the formulations showed a characteristic crystallization peak at ~40 °C. The structural reorganization and crystal growth due to the addition of lecithin affected its mechanical property significantly. The spreadability test among all prepared oleogels showed better spreadable properties for Sf5 and Sy1 oleogel. The inclusion of lecithin in oleogels has demonstrated an enhancement in oleogel properties that allows them to be included in various food products.

Polysaccharide-Based Nanocomposites for Food Packaging Applications.

The article presents a review of the literature on the use of polysaccharide bionanocomposites in the context of their potential use as food packaging materials. Composites of this type consist of at least two phases, of which the outer phase is a polysaccharide, and the inner phase (dispersed phase) is an enhancing agent with a particle size of 1-100 nm in at least one dimension. The literature review was carried out using data from the Web of Science database using VosViewer, free software for scientometric analysis. Source analysis concluded that polysaccharides such as chitosan, cellulose, and starch are widely used in food packaging applications, as are reinforcing agents such as silver nanoparticles and cellulose nanostructures (e.g., cellulose nanocrystals and nanocellulose). The addition of reinforcing agents improves the thermal and mechanical stability of the polysaccharide films and nanocomposites. Here we highlighted the nanocomposites containing silver nanoparticles, which exhibited antimicrobial properties. Finally, it can be concluded that polysaccharide-based nanocomposites have sufficient properties to be tested as food packaging materials in a wide spectrum of applications.

Selected Applications of Chitosan Composites.

Chitosan is one of the emerging materials for various applications. The most intensive studies have focused on its use as a biomaterial and for biomedical, cosmetic, and packaging systems. The research on biodegradable food packaging systems over conventional non-biodegradable packaging systems has gained much importance in the last decade. The deacetylation of chitin, a polysaccharide mainly obtained from crustaceans and shrimp shells, yields chitosan. The deacetylation process of chitin leads to the generation of primary amino groups. The functional activity of chitosan is generally owed to this amino group, which imparts inherent antioxidant and antimicrobial activity to the chitosan. Further, since chitosan is a naturally derived polymer, it is biodegradable and safe for human consumption. Food-focused researchers are exploiting the properties of chitosan to develop biodegradable food packaging systems. However, the properties of packaging systems using chitosan can be improved by adding different additives or blending chitosan with other polymers. In this review, we report on the different properties of chitosan that make it suitable for food packaging applications, various methods to develop chitosan-based packaging films, and finally, the applications of chitosan in developing multifunctional food packaging materials. Here we present a short overview of the chitosan-based nanocomposites, beginning with principal properties, selected preparation techniques, and finally, selected current research.

Fabrication and Characterization of Poly (vinyl alcohol) and Chitosan Oligosaccharide-Based Blend Films.

In the present study, we report the development of poly (vinyl alcohol) (PVA) and chitosan oligosaccharide (COS)-based novel blend films. The concentration of COS was varied between 2.5-10.0 wt% within the films. The inclusion of COS added a brown hue to the films. FTIR spectroscopy revealed that the extent of intermolecular hydrogen bonding was most prominent in the film that contained 5.0 wt% of COS. The diffractograms showed that COS altered the degree of crystallinity of the films in a composition-dependent manner. As evident from the thermal analysis, COS content profoundly impacted the evaporation of water molecules from the composite films. Stress relaxation studies demonstrated that the blend films exhibited more mechanical stability as compared to the control film. The impedance profiles indicated the capacitive-dominant behavior of the prepared films. Ciprofloxacin HCl-loaded films showed excellent antimicrobial activity against Escherichia coli and Bacillus cereus. The prepared films were observed to be biocompatible. Hence, the prepared PVA/COS-based blend films may be explored for drug delivery applications.

Graphene Oxide Increases Corneal Permeation of Ciprofloxacin Hydrochloride from Oleogels: A Study with Cocoa Butter-Based Oleogels.

In this work, oleogels of cocoa butter (CB), rice bran oil (RBO), and graphene oxide (GO) were prepared. The prepared oleogels were subjected to various characterization techniques such as bright-field microscopy, X-ray diffraction (XRD), crystallization kinetics, differential scanning calorimetry (DSC), and mechanical studies. The influence of increasing GO content on the in vitro drug release and ex vivo corneal permeation of the model drug (ciprofloxacin HCl-CPH) from the oleogels was also investigated. Bright-field micrographs showed that increment in GO content reduced the size of the globular particles of CB. XRD analysis revealed that CB was crystallized in its ß' and ß polymorphic forms in the oleogels, which was in agreement with thermal studies. The mechanical characterization demonstrated that the presence of GO improved the elastic nature and stress-bearing properties of the oleogels. Moreover, GO altered the crystallization kinetics of CB in the oleogels in a composition-dependent manner. The in vitro release of CPH from the oleogels occurred through either Fickian diffusion or fat network relaxation or a combination thereof. Furthermore, the inclusion of GO enhanced the ex vivo permeation of CPH molecules across the caprine cornea. Hence, we concluded that the prepared oleogels could be explored as potential delivery systems for ophthalmic applications.

Synthesis and characterization of novel tamarind gum and rice bran oil-based emulgels for the ocular delivery of antibiotics.

In this study, we developed tamarind gum (TG) and rice bran oil (RBO)-based emulgels. The control formulation (TR0), did not contain RBO. The emulgels were named as TR1, TR2, TR3, and TR4, which contained 5% (w/w), 10% (w/w), 15% (w/w), and 20% (w/w/) of RBO, respectively. The microscopic studies showed that the emulgels were biphasic in nature. FTIR spectroscopy revealed the reduction in the hydrogen bonding with an increase in the RBO content. Impedance profiles suggested that the resistive component of the emulgels was increased as the RBO content was increased. The thermal analysis suggested that the addition of RBO reduced the water holding capacity of the emulgels. Stress relaxation studies revealed that the fluidic component was considerably higher in TG/RBO-based emulgels as compared to TR0. In vitro release study of the model drug (ciprofloxacin HCl; a hydrochloride salt of ciprofloxacin) suggested a significantly lower release from the emulgel matrices (TR1-TR4) in comparison to TR0. However, ex vivo corneal permeation of the drug increased with an increase in the RBO content. Since the emulgels were able to improve the corneal permeation of the model drug, the emulgels can be explored to deliver drugs to the internal structures of the eye.

Synthesis, characterization, and antimicrobial efficacy of composite films from guar gum/sago starch/whey protein isolate loaded with carvacrol, citral and carvacrol-citral mixture.

The aim of this research was to formulate antimicrobial, composite films of guar gum, sago starch, and whey protein isolate for the prophylaxis of the bacterial gastroenteritis. The model antibacterial agents incorporated were essential oils, namely, carvacrol, citral and their combination. The films became darker and brownish in color due to the entrapment of the oils. The surface of the oil-entrapped films was more rough and coarse compared to the control film. Confocal micrographs affirmed the uniform distribution of the oil droplets within the biopolymeric network. The highest crystallite size and lowest lattice strain were estimated in the citral-containing film. FTIR analysis demonstrated that the incorporation of citral increased the proportion of the ß-sheet structures of the whey protein isolate within the film matrix. However, the film formulation containing combination of carvacrol and citral demonstrated the lowest water vapor transmission rate (WVTR), highest tensile strength, Young's modulus and work to failure. All the oil-containing films demonstrated good antibacterial potency against the model bacterial gastroenteritis causing bacteria, namely, Bacillus cereus and Escherichia coli. In gist, it can be concluded that the prepared antimicrobial films could be used for the prophylaxis of the bacterial gastroenteritis.

Composite edible films and coatings from food-grade biopolymers.

The development of edible films and coatings from food-grade biopolymers has advanced significantly during the past decade. The current state-of-the-art lies in the formulation of composite edible films and coatings from such biomolecules. Composite films and coatings refer to systems where multiple biopolymers have been combined to achieve beneficial properties. Carbohydrate, protein, and lipids have been preferred for developing such systems. Binary films and coatings have been prepared from multiple combinations including protein-protein, carbohydrate-carbohydrate and protein-carbohydrate. Similarly, ternary films and coatings have been prepared from protein-protein-carbohydrate combinations. In addition, several active ingredients including antimicrobial compounds have been loaded to these systems for the preparation of functional films and coatings. Therefore, the goal of this manuscript is to review the multitude of composite systems that are currently available for food packaging purposes. In addition, we discuss the application of composite coatings to fruits and vegetables, dairy, meat and seafood as model food systems.

Multiple layers and conjugate materials for food emulsion stabilization.

A single emulsifier material is seldom sufficient to cater to the requirements of complex emulsion-based food formulations that have to function over a wide range of pH, ionic strength, storage time, and temperature. Food emulsions have to be designed to satisfy several requirements for use which could be achieved by combining the beneficial properties of multiple emulsifiers. The present article reviews novel biological materials that are used to design oil-in-water (o/w) emulsions. More specifically, the major focus is to discuss (o/w) interfaces stabilized by multiple biopolymers. Prevalent ways by which two or more emulsifiers could be combined is by (i) forming multi-layered emulsions and (ii) conjugation of two compounds of beneficial traits. Multilayer emulsions make use of a combination of protein, phospholipids, and carbohydrates to stabilize (o/w) interfaces. On the other hand, covalent bonding between a protein and polysaccharide is induced to form a single entity known as conjugate that is superior to the individual biopolymers in terms of emulsion stability. Therefore, properties required to maintain emulsion stability such as surface activity, solubility, steric and electrostatic repulsion, and antioxidant effects from two different hydrocolloids could be integrated.

Impact of starch-based emulsions on the antibacterial efficacies of nisin and thymol in cantaloupe juice.

The use of antimicrobial compounds to prevent foodborne pathogens from contaminating fresh-cut produce has received broad attentions; however, the applications of these compounds are hindered by their rapid depletion in foods. To prolong their efficacies, the use of delivery systems is essential. In this study, oil-in-water emulsions formed using starch octenyl succinate (starch-OS) were used to stabilize nisin and thymol in cantaloupe juice-containing fluid. Listeria monocytogenes V7 and Salmonella enterica serovar Typhimurium were used as model pathogens to evaluate the antimicrobial activities of nisin and thymol formulations in cantaloupe juice. The results showed that the emulsions had much greater capability to retain nisin and thymol over the storage and displayed much greater effect to inhibit Listeria and Salmonella than non-emulsion, aqueous formulations. Starch-OS based emulsions not only retained nisin and thymol activities separately, but also exhibited their cooperative antibacterial effects.

Emulsion Stabilized with Starch Octenyl Succinate Prolongs Nisin Activity Against Listeria Monocytogenes in a Cantaloupe Juice Model.

Food safety related to fresh and fresh-cut produce, such as cantaloupe, is a great challenge in the food industry. Various processing technologies, including the use of antimicrobial materials, have been applied to reduce pathogens. Nisin, an antimicrobial peptide, shows strong inhibitory activity against Listeria monocytogenes (LM); however, it suffers from quick depletion in foods. In this study, the protective effect of starch octenyl succinate (starch-OS) stabilized emulsion on nisin was evaluated in a cantaloupe juice model. Results showed that the proteolytic enzyme in cantaloupe juice caused nisin depletion, and the adsorption of nisin in emulsion led to considerable protection of antimicrobial activity. After 1, 3, and 6 d of incorporation with cantaloupe juice, the size of LM inhibitory ring were 7.64, 7.60, and 2.97 mm, respectively for emulsion formulations, whereas it reduced from 3.60 mm to a negligible level for nisin-alone. This study showed the potential of using carbohydrate colloidal systems to prolong the efficacy of antimicrobial materials.