Search Results (15)

Using free microorganisms for industrial processes has some limitations, such as the extensive consumption of substrates for growth, significant sensitivity to the microenvironment, and the necessity of separation from the product and, therefore, the cyclic process. It is widely acknowledged that confining or immobilizing cells in a matrix or support structure enhances enzyme stability, facilitates recycling, enhances rheological resilience, lowers bioprocess costs, and serves as a fundamental prerequisite for large-scale applications. This report summarizes the various cell immobilization methods, including several synthetic (polyvinylalcohol, polyethylenimine, polyacrylates, and Eudragit) and natural (gelatin, chitosan, alginate, cellulose, agar–agar, carboxymethylcellulose, and other polysaccharides) polymeric materials in the form of thin films, hydrogels, and cryogels. Advancements in the production of well-known antibiotics like penicillin and cephalosporin by various strains were discussed. Additionally, we highlighted cutting-edge research related to strain producers of peptide-based antibiotics (polymyxin B, Subtilin, Tyrothricin, varigomycin, gramicidin S, friulimicin, and bacteriocin), glusoseamines, and polyene derivatives. Crosslinking agents, especially covalent linkers, significantly affect the activity and stability of biocatalysts (penicillin G acylase, penicillinase, deacetoxycephalosporinase, L-asparaginase, β-glucosidase, Xylanase, and urease). The molecular weight of polymers is an important parameter influencing oxygen and nutrient diffusion, the kinetics of hydrogel formation, rigidity, rheology, elastic moduli, and other mechanical properties crucial for long-term utilization. A comparison of stability and enzymatic activity between immobilized enzymes and their free native counterparts was explored. The discussion was not limited to recent advancements in the biopharmaceutical field, such as microorganism or enzyme immobilization, but also extended to methods used in sensor and biosensor applications. In this study, we present data on the advantages of cell and enzyme immobilization over microorganism (bacteria and fungi) suspension states to produce various bioproducts and metabolites—such as antibiotics, enzymes, and precursors—and determine the efficiency of immobilization processes and the optimal conditions and process parameters to maximize the yield of the target products. Full article

Walnuts play a positive role in human health due to their large amounts of unsaturated fatty acids, whereas lipid oxidation can easily occur during storage. Herein, three natural antioxidants (epicatechin, sesamol, and myricetin) were added to the composite film cross-linked with chitosan and soy protein peptide, and the antioxidant film appropriate for the preservation of walnut kernels from Juglans sigillata was screened to improve the storage quality of walnuts. The results showed that three antioxidant films could all enhance the storage performance of walnut kernels, with sesamol being the best. The characterization of antioxidant film cross-linked with chitosan and soy protein peptide containing sesamol (C/S-ses film) revealed that the composite film improved the slow release and stability of sesamol; in addition, the presence of sesamol could effectively reduce the light transmittance and water vapor permeability of the composite film, together with significantly enhancing the antioxidant and antimicrobial activities, resulting in an effective prolongation of the storage period of walnut kernels. These findings indicated that C/S-ses possess excellent potential for retarding the oxidative rancidity of unsaturated fatty acids and will provide an effective strategy for the preservation of walnut kernels. Full article

Southern King Crab (SKC) represents an important fishery resource that has the potential to be a natural source of chitosan (CS) production. In tissue engineering, CS is very useful to generate biomaterials. However, CS has a lack of signaling molecules that facilitate cell–substrate interaction. Therefore, RGD (arginine–glycine–aspartic acid) peptides corresponding to the main integrin recognition site in extracellular matrix proteins have been used to improve the CS surface. The aim of this study was to evaluate in vitro cell adhesion and proliferation of CS films synthesized from SKC shell wastes functionalized with RGD peptides. The FTIR spectrum of CS isolated from SKC shells (SKC-CS) was comparable to commercial CS. Thermal properties of films showed similar endothermic peaks at 53.4 and 53.0 °C in commercial CS and SKC-CS, respectively. The purification and molecular masses of the synthesized RGD peptides were confirmed using HPLC and ESI-MS mass spectrometry, respectively. Mouse embryonic fibroblast cells showed higher adhesion on SKC-CS (1% w/v) film when it was functionalized with linear RGD peptides. In contrast, a cyclic RGD peptide showed similar adhesion to control peptide (RDG), but the highest cell proliferation was after 48 h of culture. This study shows that functionalization of SKC-CS films with linear or cyclic RGD peptides are useful to improve effects on cell adhesion or cell proliferation. Furthermore, our work contributes to knowledge of a new source of CS to synthesize constructs for tissue engineering applications. Full article

It is key to fight bacterial adhesion to prevent biofilm establishment on biomaterials. Surface immobilization of antimicrobial peptides (AMP) is a promising strategy to avoid bacterial colonization. This work aimed to investigate whether the direct surface immobilization of Dhvar5, an AMP with head-to-tail amphipathicity, would improve the antimicrobial activity of chitosan ultrathin coatings. The peptide was grafted by copper-catalyzed azide-alkyne cycloaddition (CuAAC) chemistry by either its C- or N- terminus to assess the influence of peptide orientation on surface properties and antimicrobial activity. These features were compared with those of coatings fabricated using previously described Dhvar5-chitosan conjugates (immobilized in bulk). The peptide was chemoselectively immobilized onto the coating by both termini. Moreover, the covalent immobilization of Dhvar5 by either terminus enhanced the antimicrobial effect of the chitosan coating by decreasing colonization by both Gram-positive (Staphylococcus aureus, Staphylococcus epidermidis) and Gram-negative (Escherichia coli, Pseudomonas aeruginosa) bacteria. Relevantly, the antimicrobial performance of the surface on Gram-positive bacteria depended on how Dhvar5-chitosan coatings were produced. An antiadhesive effect was observed when the peptide was grafted onto prefabricated chitosan coatings (film), and a bactericidal effect was exhibited when coatings were prepared from Dhvar5-chitosan conjugates (bulk). This antiadhesive effect was not due to changes in surface wettability or protein adsorption but rather depended on variations in peptide concentration, exposure, and surface roughness. Results reported in this study show that the antibacterial potency and effect of immobilized AMP vary greatly with the immobilization procedure. Overall, independently of the fabrication protocol and mechanism of action, Dhvar5-chitosan coatings are a promising strategy for the development of antimicrobial medical devices, either as an antiadhesive or contact-killing surface. Full article

There is a significant drive towards the development of edible biocompatible films for food packaging application due to the environmental and health impacts of synthetic packaging materials. This has inspired the exploration of biodegradable natural polymers as packaging materials. To address the instant water disintegration of most natural polymers, polymers with conditional water solubility, such as chitosan (needing acidic conditions for dissolution in water), have gained significant research attention. To this end, chitosan has been blended with different natural proteins, including whey protein isolates, to prepare edible food films. However, consumption of whey protein isolates in their natural form has been proposed in the literature to prolong processing (digestion) time upon consumption. To circumvent this limitation, here we report the development of chitosan/whey protein hydrolysate-based edible films with additional antioxidant properties. The developed films revealed that the inclusion of whey protein hydrolysate improved physicochemical properties and mechanical strength of the films with tensile strength of 26.3 MPa at 1 wt% WPH loading compared to 10.9 MPa in control neat chitosan films (0 wt% WPH). Furthermore, chitosan/whey protein hydrolysate exhibited a significant (whey protein hydrolysate) dose-dependent antioxidant response with a maximum value of 83% DPPH in chitosan/WPH (1 wt%) films assessed using two different antioxidant assays. Based on the results from this study, we envisage the exploration of whey protein hydrolysate-based films for commercial food packaging application in future. Full article

In this study, layer-by-layer assembly was performed to prepare sodium alginate (SA) layer and walnut-peptide–chitosan (CS) bilayer composite films. Genipin was adopted to crosslink CS and walnut peptide. The properties of walnut peptide-CS-SA composite film were determined, and the influence of material ratio on the performance of composite film was explored. According to the results, the mechanical tensile property, oil absorption property, and water vapor barrier property of the composite film were improved with the presence of genipin. Moreover, the proportion of CS and walnut peptide had significant effects on color, transmittance, mechanical properties, barrier properties, and antioxidant properties of the composite films. Among them, the composite film containing 1% (w/v) CS, 1% (w/v) walnut peptide, and 0.01% (w/v) genipin showed the best performance, with a tensile strength of 3.65 MPa, elongation at break of 30.82%, water vapor permeability of 0.60 g·mm·m⁻²·h⁻¹·kPa⁻¹, oil absorption of 0.85%, and the three-phase electrochemistry of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging rate of 25.59%. Under this condition, the tensile property, barrier property, and oxidation resistance of the composite film are good, which can provide a good preservation effect for food, and has great application potential. Full article

The sublingual mucosa is an attractive route for drug delivery, although challenged by a continuous flow of saliva that leads to a loss of drug by swallowing. It is of great benefit that drugs absorbed across the sublingual mucosa avoid exposure to the harsh environment of the gastro-intestinal lumen; this is especially beneficial for drugs of low physicochemical stability such as therapeutic peptides. In this study, a two-layered hybrid drug delivery system was developed for the sublingual delivery of the therapeutic peptide desmopressin. It consisted of peptide-loaded mucoadhesive electrospun chitosan/polyethylene oxide-based nanofibers (mean diameter of 183 ± 20 nm) and a saliva-repelling backing film to promote unidirectional release towards the mucosa. Desmopressin was released from the nanofiber-based hybrid system (approximately 80% of the loaded peptide was released within 45 min) in a unidirectional manner in vitro. Importantly, the nanofiber–film hybrid system protected the peptide from wash-out, as demonstrated in an ex vivo flow retention model with porcine sublingual mucosal tissue. Approximately 90% of the loaded desmopressin was retained at the surface of the ex vivo porcine sublingual mucosa after 15 min of exposure to flow rates representing salivary flow. Full article

Background: Biological molecules such as therapeutic proteins and peptides have provided ample opportunities for further improvements in health and wellbeing. However, such biomacromolecules face stability challenges regarding premature processing and digestion, whenever any enteric delivery route is considered. Hence, oral guar films entrapping peptide-containing chitosan microparticles have been developed as a new delivery system to carry and protect therapeutic relaxing peptides. Results: The composite films were characterized regarding physicochemical properties using FTIR as well as peptide encapsulation efficiency, cytotoxicity, and peptide permeability through buccal and intestinal cell-based models. The composite films with 0.18% (w/w) of peptide demonstrated ca. 0.63% of peptide loading efficiency into the chitosan microparticles and to be safe for buccal and intestinal epithelial cells up to 24 h of contact. The permeability assays through TR146 cells to mimic buccal mucosa epithelium and Caco-2/HT29-MTX to mimic duodenum conditions showed that the incorporation of peptide chitosan microparticles into guar-gum oral films offered a faster permeability, when compared with the free peptide and peptide-chitosan-microparticles-loaded chitosan. Conclusion: This study suggests that peptide-loaded chitosan microparticles into guar-gum oral films might be a promising carrier for enhancement of the buccal and intestinal absorption of a relaxing peptide. Full article

The high mortality rate of different multi-resistant bacteria (MDR) has led to an immediate and urgent solution. Patients hospitalized for chronic diseases have a weakened immune system and are at high risk of contracting an opportunistic infection. Likewise, the WHO prioritized studies against a selected group of MDR bacteria for their control [¹]. In this scope, the Ctx(Ile²¹)-Ha antimicrobial peptide (AMP) presented great potential and efficient biological activity against Acinetobacter baumannii and Pseudomonas aureginosa MDR bacteria [²,³,⁴]. Thus, the aim of this research was to design ultrasound-assisted micro-structured films loaded with the Ctx(Ile²¹)-Ha AMP, based on starch and chitosan, for its effective protective action. Gelling was performed for grain breaking and to expose the hydroxyls [5]. For this, 10 g of cornstarch was used as well as 300 mL of distilled water under agitation at 90 °C for 1 h. Then, 5 mL of the gelled starch was added and mixed with 50 mg of peptide. Then, it was stored in petri dishes at 50 °C for 5 h. Chitosan film was synthesized by free-radical polymerization in the presence of crosslinker [⁶]. Chitosan dispersion (CD) was prepared by dissolving 2% w/v chitosan in 2% v/v acetic acid solution. Ctx(Ile²¹)-Ha was placed on the CD with 0.3% w/v of glycerol and magnetic agitation at 150 rpm. For this, its properties were evaluated by DSC/TGA, FTIR, XRD, and SEM. The physicochemical stability studies of the AMP showed its structure unchanged for up to 3 months exposed to water and for up to one year in the form of a dry film. These results were confirmed by the LC/MS profile, in which XDR indicates a consistent semi-morpho phase. Finally, with these results, we check its stability and protective potential over time and, based on previously published results on their activity against MDR bacteria [²], we conclude that the new products based on AMPs could be potential anti-MDR bacterial agents, avoiding the exposure of critically ill patients in intensive care or post-surgery beds and preventing their dissemination. Full article

Active films based on carboxymethyl chitosan incorporated corn peptide were developed, and the effect of the concentration of corn peptide on films was evaluated. Physicochemical properties of the films, including thickness, opacity, moisture content, color, mechanical properties, water vapor permeability, and oil resistance, were measured. Biological activities of the films, including the antioxidant and antibacterial activities, were characterized in terms of 2, 2-diphenyl-1-picrylhydrazyl free radical scavenging activity, reducing power, the total antioxidant activity, and the filter disc inhibition zone method. The results indicated that the incorporation of corn peptide caused interactions between carboxymethyl chitosan and corn peptide in Maillard reaction and gave rise to the films light yellow appearance. Compared with the Control, the degree of glycosylation, browning intensity, thickness, opacity, tensile strength, antioxidant activity, and antibacterial activity of films were increased, but the elongation, vapor permeability, and oil resistance of films were decreased. The films based on corn peptide and carboxymethyl chitosan can potentially be applied to food packaging. Full article

The therapeutic use of peptides has increasingly recognized in the development of new therapies. However, the susceptible enzymatic cleavage is a barrier that needs to overcome. Nose-to-brain delivery associated with liposomes can protect peptides against biodegradation and improve the accessibility to brain targets. The aim was to develop a liposomal formulation as ghrelin carrier. The quality by design (QbD) approach was used as a strategy for method development. The initial risk assessments were carried out using a fishbone diagram. A screening design study was performed for the critical material attributes/critical process parameters (CMAs/CPPs) on critical quality attributes (CQAs). Liposomes were obtained by hydrating phospholipid films, followed by extrusion or homogenization, and coated with chitosan. The optimized liposome formulation was produced by high-pressure homogenization coated with chitosan, and the resulted were liposomes size 72.25 ± 1.46 nm, PDI of 0.300 ± 0.027, the zeta potential of 50.3 ± 1.46 mV, and encapsulation efficiency of 53.2%. Moreover, chitosan coating improved performance in ex vivo permeation and mucoadhesion analyzes when compared to the uncoated liposome. In this context, chitosan coating is essential for the performance of the formulations in the ex vivo permeation and mucoadhesion analyzes. The intranasal administration of ghrelin liposomes coated with chitosan offers an innovative opportunity to treat cachexia. Full article

Transdermal drug delivery is used to deliver a drug by eliminating the first-pass metabolism, which increases the bioavailability of the drug. The present study aims to formulate the chitosan—tamarind seed polysaccharide composite films and evaluate for the delivery of protein/peptide molecules. Nine formulations were prepared and evaluated by using different parameters, such as physical appearance, folding endurance, thickness of film, surface pH, weight variation, drug content, surface morphology, percentage moisture intake and uptake, drug release kinetics, and drug permeability. The film weight variance was observed between 0.34 ± 0.002 to 0.47 ± 0.003 g. The drug level of the prepared films was found to be between 96 ± 1.21 and 98 ± 1.33μg. Their intake of moisture ranged between 2.83 ± 0.002 and 3.76 ± 0.001 (%). The moisture absorption of the films ranged from 5.33 ± 0.22 to 10.02 ± 0.61 (%). SEM images revealed a smooth film surface, while minor cracks were found in the film after permeation tests. During the first 4 days, drug release was between 13.75 ± 1.64% and 22.54 ± 1.34% and from day 5 to day 6, it was between 72.67 ± 2.13% and 78.33 ± 3.13%. Drug permeation during the first 4 days was 15.78 ± 1.23 %. Drug permeation (%) during the first 4 days was between 15.78 ± 1.23 and 22.49 ± 1.29 and from day 5 to day 6, it was between 71.49 ± 3.21 and 77.93 ± 3.20. Full article

Among different insects, the American cockroach (Periplaneta americana) has been bred in industrial scale successfully as a potential resource of protein, lipid, and antibacterial peptide. However, the application of its chitosan has not been studied widely, which has hindered the sufficient utilization of P. americana. In this paper, the chitosan from P. americana was separated, characterized, and processed into film (PaCSF) to examine its potential of being applied in food packaging. As the results of different characterizations showed, PaCSF was similar to shrimp chitosan film (SCSF). However, concerning the performances relating to food packaging, the two chitosan films were different. PaCSF contained more water (42.82%) than SCSF did, resulting in its larger thickness (0.08 mm). PaCSF could resist UV light more effectively than SCSF did. Concerning antioxidant activity, the DPPH radical scavenging ability of PaCSF increased linearly with time passing, reaching 72.46% after 8 h, which was better than that of SCSF. The antibacterial activity assay exhibited that PaCSF resisted the growth of Serratia marcescens and Escherichia coli more effectively than SCSF did. The results implied that P. americana chitosan could be a potential raw material for food packaging, providing a new way to develop P. americana. Full article

Chitosan/peptide films were prepared by incorporating peptides (0.4%, w/v) from soy, corn and caseins into chitosan films. The presence of peptides significantly affected the physical, antibacterial and antioxidative properties of chitosan films. Among these films, those containing corn peptide showed the best water vapor barrier properties, and the tensile strength and elongation at break increased to 24.80 Mpa and 23.94%, respectively. Characterization of surface hydrophobicity and thermal stability suggested the strongest intermolecular interactions between corn peptides and chitosan. Moreover, films containing casein peptides showed the highest antibacterial activity and radical scavenging activity. The DPPH scavenging rate of films containing casein peptides reached 46.11%, and ABTS scavenging rate reached 66.79%. These results indicate the chitosan/peptide films may be promising food packaging materials. Full article

Nowadays, bioactive peptides are used for therapeutic applications and the selection of a carrier to deliver them is very important to increase the efficiency, absorption, release, bioavailability and consumer acceptance. The aim of this study was to develop and characterize chitosan-based films loaded with chitosan microparticles containing a bioactive peptide (sequence: KGYGGVSLPEW) with antihypertensive properties. Films were prepared by the solvent casting method, while the microparticles were prepared by ionic gelation. The final optimized chitosan microparticles exhibited a mean diameter of 2.5 µm, a polydispersity index of 0.46, a zeta potential of +61 mV and a peptide association efficiency of 76%. Chitosan films were optimized achieving the final formulation of 0.79% (w/v) of chitosan, 6.74% (w/v) of sorbitol and 0.82% (w/v) of citric acid. These thin (±0.100 mm) and transparent films demonstrated good performance in terms of mechanical and biological properties. The oral films developed were flexible, elastic, easy to handle and exhibited rapid disintegration (30 s) and an erosion behavior of 20% when they came into contact with saliva solution. The cell viability (75–99%) was proved by methylthiazolydiphenyl-tetrazolium bromide (MTT) assay with TR146 cells. The chitosan mucoadhesive films loaded with peptide–chitosan microparticles resulted in an innovative approach to perform administration across the buccal mucosa, because these films present a larger surface area, leading to the rapid disintegration and release of the antihypertensive peptide under controlled conditions in the buccal cavity, thus promoting bioavailability. Full article