Nelumbo nucifera flower extract incorporated alginate/polyvinyl alcohol films as a sustainable pH indicator for active food packaging applications.

In recent years, there has been a growing demand for environmentally friendly smart packaging materials. Therefore, in this study, we developed an eco-friendly pH-sensitive indicator film through the solvent casting process, incorporating alginate, polyvinyl alcohol, garlic, and Nelumbo nucifera flower extract. The effect of extract on the chemical and physical properties of the film were extensively studied using various characterization techniques. XRD and FTIR reveal the strong interaction between the polymers and the extract. The incorporation of the extract influenced various parameters such as swelling behavior, water solubility, and moisture content, while also improving the film's thermal stability, biodegradability, as well as its antioxidant and antimicrobial properties. Interestingly, the film exhibited a color change in response to pH change. During shrimp storage, the film showed a visible transition from purple to green, indicating shrimp spoilage. Additionally, the film's ability to detect freshness was confirmed by measuring total volatile basic nitrogen (TVBN). These findings suggest that the PVA/alginate/garlic/Nelumbo nucifera film shows promise as an intelligent packaging material for real-time food monitoring applications.

Effect of carbon dot-doped Ti-MOF on CMC/Agar film and active packaging application on storage quality of fruits.

Ti-metal organic framework (Ti-MOF) doped with carbon dots (CDs) with enhanced antibacterial potential was synthesized using solvothermal-assisted mechanical stirring and used for the fabrication of CMC/Agar-based active packaging films. The incorporation of CD@Ti-MOF not only improved the tensile strength of the CMC/Agar film by 17.4% but also exhibited strong antioxidant activity with 100% of ABTS and 57.8% of DPPH radical scavenging using 0.64 cm2/mL of CMC/Agar/CD@Ti-MOF film. Furthermore, water vapor permeability, oxygen permeability, and ultraviolet light-blocking ability (95.7% of UV-B and 84.7% of UV-A) were improved significantly. The CMC/Agar/CD@Ti-MOF film showed strong antibacterial activity and could inhibit the progress of E. coli up to 8.2 Log CFU/mL and completely stopped the growth of L.monocytogenes after 12 h of incubation. Additionally, CMC/Agar/CD@Ti-MOF film extended the shelf life of cherry tomatoes preserved at 4 °C and delayed the quality degradation, maintaining the visual aspects of the packaging.

Carrageenan-based functional films hybridized with carbon dots and anthocyanins from rose petals for smart food packaging applications.

Multifunctional smart biopolymeric films were fabricated using rose petal anthocyanin (RPA) and carrageenan (CAR) doped with rose petal-derived carbon dots (RP-CDs). Response surface-optimized RPA showed the highest total anthocyanins and radical scavenging ability. Produced RP-CD exhibited UV absorption and high fluorescence with antibacterial/antioxidant abilities. Enrichment with 2 % RP-CD and 5 % RPA in the CAR matrix results in improved physicochemical, i.e., water contact angle, water vapor permeability, and UV-blocking properties of the fabricated material. Results showed that nanocomposite films scavenged radicals better than the neat CAR films. Zeta potential, FTIR, SEM, and XPS suggested improved compatibility/stability and enhanced elemental configuration of RP-CDs/RPA additives in the CAR polymer matrix. Perishable food packaging (minced pork and shrimp) demonstrated that nanocomposite films work efficiently and non-destructively and are promising tools for monitoring real-time freshness through interpretable visual changes from red to yellow. The CAR/RP-CDs/RPA-based nanocomposite indicator films are expected to be applied as various smart packaging materials. These films possess the ability to promptly detect changes in quality, preserve the quality, and prolong the shelf life of packaged foods.

Carboxymethyl cellulose/gelatin film incorporated with eggplant peel waste-derived carbon dots for active fruit packaging applications.

Carbon dots (CDs) were derived using eggplant peel by a hydrothermal approach and incorporated into the carboxymethyl cellulose (CMC) and gelatin (Gel) blend to develop sustainable and functional packaging films for fruit preservation. The CD was uniformly dispersed within the CMC/Gel blend to form a dense and continuous film and fashioned a strong interaction with the polymer chain, increasing the tensile strength of the film by 5.0-16.0 %. Also, with the impregnation of CDs, the UV-blocking potential of the CMC/Gel film was greatly improved to the extent of blocking 94.3 % of UV-B and 72.5 % of UV-A, while the water vapor permeability slightly decreased (by 2.7-5.4 %), and the water contact angle of the film marginally expand (by 6.2-19.1 %). The CMC/Gel film with 3 wt% of CD added depicted strong antioxidant efficacy of 100 % against ABTS and 59.1 % against DPPH and displayed strong antibacterial action that inhibited the progress of Listeria monocytogenes and Escherichia coli by 99.8 %. In addition, when table grapes were packaged using a CMC/Gel composite film containing CD and stored at 4 °C for 24 days, the fruits packed with the composite film maintained excellent external quality and extended the shelf life.

Development of smart colorimetric indicators for tracking kimchi freshness by loading aronia extract in agar, κ-carrageenan, and cellulose nanofiber films.

Color indicator films incorporating aronia extract powder (AEP) and biopolymers like agar, carrageenan, and cellulose nanofiber (CNF) were developed to monitor kimchi freshness. AEP-containing films showed strong UV-barrier properties, and reduced light transmittance by 99.12 % for agar, 98.86 % for carrageenan, and 98.67 % for CNF-based films. All AEP-films exhibited high sensitivity to pH changes and vapor exposure to ammonia and acetic acid. Color change notably influenced by the polymer type, particularly evident with ammonia vapor exposure, especially in the AEP/carrageenan film. The chemical structure and thermal stability of the biopolymers remained unchanged after AEP-addition. Tensile strength increased by 24.2 % for AEP/CNF but decreased by 19.4 % for AEP/agar and 24.3 % for AEP/carrageenan films. AEP-containing films displayed strong antioxidant activity, with 99 % free radical scavenging in ABTS and ~ 80 % in DPPH assays. Alkalized AEP-indicator films were more effective in detecting color changes during kimchi packaging tests. Among the labels, alkalized AEP/agar film showed the most obvious color change from green-gray (fresh kimchi, pH 5.5, acidity 0.48 %) to pale brown (optimal fermentation, pH 4.6, acidity 0.70 %), and pale violet-brown (over-fermented, pH 3.80, acidity 1.35 %). Alkalized AEP-indicator films offer promising real-time detection of packed fermented foods like kimchi.

Active Fish Gelatin/Chitosan Blend Film Incorporated with Guava Leaf Powder Carbon Dots: Properties, Release and Antioxidant Activity.

Active packaging is an innovative approach to prolonge the shelf-life of food products while ensuring their quality and safety. Carbon dots (CDs) from biomass as active fillers for biopolymer films have been introduced to improve their bioactivities as well as properties. Gelatin/chitosan (G/C) blend films containing active guava leaf powder carbon dots (GL-CDs) at various levels (0-3%, w/w) were prepared by the solvent casting method and characterized. Thickness of the control increased from 0.033 to 0.041 mm when 3% GL-CDs were added (G/C-CD-3%). Young's modulus of the resulting films increased (485.67-759.00 MPa), whereas the tensile strength (26.92-17.77 MPa) and elongation at break decreased (14.89-5.48%) as the GL-CDs' level upsurged (p < 0.05). Water vapor barrier property and water contact angle of the film were enhanced when incorporated with GL-CDs (p < 0.05). GL-CDs had a negligible impact on film microstructure, while GL-CDs interacted with gelatin or chitosan, as determined by FTIR. The release of GL-CDs from blend films was more pronounced in water than in alcoholic solutions (10-95% ethanol). The addition of GL-CDs improved the UV light barrier properties and antioxidant activities of the resultant films in a dose-dependent manner. Thus, GL-CD-added gelatin/chitosan blend films with antioxidant activities could be employed as potential active packaging for the food industry.

Development of innovative active packaging films using gelatin/pullulan-based composites incorporated with cinnamon essential oil-loaded metal-organic frameworks for meat preservation.

This study aimed to investigate the effect of cinnamon essential oil (CEO)-loaded metal-organic frameworks (CEO@MOF) on the properties of gelatin/pullulan (Gel/Pull)-based composite films (Gel/Pull-based films). The incorporation of CEO@MOF into Gel/Pull-based films demonstrated significant antimicrobial activity against S. aureus, S. enterica, E. coli, and L. monocytogenes. Additionally, CEO@MOF integrated film exhibited a 98.16 % ABTS radical scavenging, with no significant change in the mechanical properties of the neat Gel/Pull film. The UV blocking efficiency of the composite films increased significantly from 81.38 to 99.56 % at 280 nm with the addition of 3 wt% CEO@MOF. Additionally, Gel/Pull/CEO@MOF films effectively extended the shelf life of meat preserved at 4 °C by reducing moisture loss by 3.35 %, maintaining the pH within the threshold limit (6.2), and inhibiting bacterial growth by 99.9 %. These results propose that CEO@MOF has significant potential as an effective additive in active packaging to improve shelf life and food safety.

Cellulose nanofiber-based multifunctional composite films integrated with zinc doped-grapefruit peel-based carbon quantum dots.

This study aimed to develop a multifunctional active composite film to extend the shelf life of minced pork. The composite film was prepared by incorporating zinc-doped grapefruit peel-derived carbon quantum dots (Zn-GFP-CD) into a cellulose nanofiber (CNF) matrix. The resulting film significantly improved UV-blocking properties from 39.0 % to 85.7 % while maintaining the film transparency. Additionally, the CNF/Zn-GFP-CD5% composite film exhibits strong antioxidant activity with ABTS and DPPH radical scavenging activities of 99.8 % and 77.4 %, respectively. The composite film also showed excellent antibacterial activity against both Gram-negative and Gram-positive bacteria. When used in minced pork packaging, the composite films effectively inhibit bacterial growth, maintaining bacterial levels below 7 Log CFU/g after 15 days and sustaining a red color over a 21-day storage period. Additionally, a significant reduction in the lipid oxidation of the minced pork was observed. These CNF/Zn-GFP-CD composite films have a great potential for active food packaging applications to extend shelf life and maintain the visual quality of packaged meat.

Storage stability of Asian seabass oil-in-water Pickering emulsion packed in pouches made from electrospun and solvent casted bilayer films from poly lactic acid/chitosan-gelatin blend containing epigallocatechin gallate.

Bilayer pouches were fabricated with chitosan (CS)-fish gelatin (FG) mixture containing epigallocatechin gallate (EGCG) deposited over the poly lactic acid (PLA) film through solvent casting and electrospinning techniques. Pickering emulsions (PE) of Asian seabass depot fat oil stabilized by zein colloidal particles were packed in bilayer pouches and stored at 28 ± 2 °C. The PE packed in pouch containing EGCG had higher emulsion and oxidative stability after 30 days of storage as witnessed by the smaller droplet size and lower values of thiobarbituric acid reactive substances, peroxide, conjugated diene and volatile compounds in comparison with control (PE packed in monolayer PLA pouch) (P < 0.05). EGCG incorporated pouch retained more linoleic acid (C18:2 n-6) and linolenic acid (C18:3 n-9) in emulsion than PLA pouch. Therefore, pouch from bilayer PLA/CS-FG films comprising EGCG could serve as active packaging and extended the shelf life of Pickering emulsion.

Sunlight-Driven Self-Cleaning Ultrafine Particulate Matter Filter with Antibacterial Activity.

Air pollution by particulate matter (PM) and airborne pathogens causes severe health problems in the human body. Presently, popular disposable air filters yield huge waste and have a fatal impact on the environment. Postuse cleaning of air filters also leads to secondary air and water pollution. Here, we report a sunlight-driven self-cleaning PM filter by coupling a full-solar-spectrum-active photocatalyst comprising up-conversion nanoparticles (UCNPs) decorated with semiconductor iron(III) oxide (UCNP@α-Fe2O3) shells stabilized upon graphene functionalized borosilicate fibrous membrane (rGO-BF). While rGO-BF ensures high PM adsorption, UCNP@α-Fe2O3 (NP) enables self-photodegradation of adsorbed PM under abundant sunlight and subsequent membrane regeneration, while preventing secondary air or water pollution. Rational surface chemistry and optimal microstructure enable our filters to remove >99% of PM2.5 under deplorable air-quality conditions. Moreover, our filter shows excellent antibacterial activity toward E. coli and S. aureus, demonstrating its potential for practical utilization in face masks, air filtering devices, and protective medical wear. This work successfully suggests an intriguing design platform for self-sustainable zero-waste air filter membranes.

Fabrication and Application of Turmeric Extract-Incorporated Oleogels Structured with Xanthan Gum and Soy Lecithin by Emulsion Template.

Turmeric extract (TE)-loaded oleogels (TE-OG) was fabricated by an emulsion template technique using xanthan gum (XG) and soy lecithin (SL) as oleogelators. The formulation for TE-OG was optimized using 0.32% XG, 1.2% SL, and 1.0% TE. The optimized TE-OG had a minimal particle size of 810.23 ± 10.68 nm as measured by the dynamic light scattering (DLS) method, and a high encapsulation efficiency (EE) of 96.62 ± 0.56%. Additionally, the optimized TE-OG exhibited a favorable zeta potential of -27.73 ± 0.44 mV, indicating the good stability of the TE-OG due to the electrostatic repulsion between particles. TE-OG formulated with 0.32% XG and 1.2% SL was subjected to frequency sweep testing to evaluate its solid-like rheological behavior. The oil-binding capacity (OBC) of TE-OG was consistently maintained above 99.99%. In vitro digestion of TE-OG demonstrated the potential of the emulsion template for controlled release, with less than 20% of the encapsulated curcumin being released in simulated gastric fluid (SGF), whereas nearly 70% was released in the simulated intestinal fluid (SIF). Moreover, TE-OG affected the rapid release of free fatty acids (FFAs), which have a positive effect on the digestion of triacylglycerols found in soybean oil (SO). TE-OG was further used as an alternative to commercial butter to produce pound cakes, and their rheological properties were compared to those of the pound cake prepared using commercial butter. The pound cake prepared using TE-OG showed a noticeable decrease in hardness from 10.08 ± 1.39 N to 7.88 ± 0.68 N and increased porosity, demonstrating the inherent capability of TE-OG to enhance the overall quality standards of bakery products.

Sustainable packaging film based on cellulose nanofibres/pullulan impregnated with zinc-doped carbon dots derived from avocado peel to extend the shelf life of chicken and tofu.

A cellulose nanofiber (CNF)/pullulan (PUL) based multifunctional composite film was developed for active packaging applications by incorporating Zn-doped avocado-derived carbon dots (Zn-ACDs). The incorporation of Zn-ACDs improved the interfacial compatibility and produced a dense cross-sectional structure of the composite films. The Zn-ACDs added film showed no significant difference in water vapor permeability and surface hydrophilicity compared to the neat CNF/PUL film, but the tensile strength and elongation at break increased by ~45.4 % and ~64.1 %, respectively. The addition of 5 wt% Zn-ACDs to the CNF/PUL matrix resulted in 100.0 % UV blocking properties, excellent antioxidant activity (100.0 % for ABTS and 68.0 % for DPPH), and complete eradication of foodborne pathogens such as Listeria monocytogenes ATCC 15313 and Escherichia coli O157:H7 after 3 h of exposure. The CNF/PUL composite film with Zn-ACDs applied to the active packaging of chicken and tofu significantly reduced the total growth of aerobic microorganisms without significantly changing the actual color of the packaged chicken and tofu for 9 days at 10°C. This study demonstrates that CNF/PUL composite films with Zn-ACDs are a sustainable and environmentally friendly option for protecting food from microbial contamination.

Carrageenan-based active and intelligent packaging films integrated with anthocyanin and TiO2-doped carbon dots derived from sweet potato peels.

Carrageenan-based sustainable active and pH-dependent color-changing composite films were fabricated by blending anthocyanin extracted from sweet potato peel (SPA) with TiO2-doped carbon dots (Ti-CDs) prepared using the biowaste of SPA extraction. The SPA and Ti-CDs were compatible with the carrageenan matrix and were uniformly dispersed in the used polymer to form a homogeneous film with increased mechanical properties. The composite film added with SPA and 3 wt% Ti-CD showed 100 % UV protection, superb antioxidant (100 % DPPH and ABTS scavenging assay), and potent antibacterial activity (complete eradication of foodborne L.monocytogenes and E. coli strains after 3 h incubation). Additionally, the composite films showed distinguishable colorimetric responses to pH 7-12 buffers and volatile ammonia. The intelligent sensing ability of the composite film was assessed through shrimp freshness monitoring, and the film's hue shifted from pink (fresh shrimp) to yellow/brown (inedible shrimp) during storage. Shrimp packaging studies have shown that composite films retard the rate of food quality change during storage and are a good indicator of shrimp spoilage. Therefore, the designed film is expected to have high applicability as a chip, and quick on-site sensor that detects seafood quality in real-time, and a highly effective multifunctional film for better product quality preservation.

Air-soil cycling of oxygenated, nitrated and parent polycyclic aromatic hydrocarbons in source and receptor areas.

Polycyclic aromatic hydrocarbons (PAHs) and their oxygenated and nitrated derivatives, OPAHs and NPAHs, are semivolatile air pollutants which are distributed and cycling regionally. Subsequent to atmospheric deposition to and accumulation in soils they may re-volatilise, a secondary source which is understudied. We studied the direction of air-soil mass exchange fluxes of 12 OPAHs, 17 NPAHs, 25 PAHs and one alkylated PAH in two rural environments being influenced by the pollutant concentrations in soil and air, by season, and by land cover. The OPAHs and NPAHs in samples of topsoil, of ambient air particulate and gas phases and in the gas-phase equilibrated with soil were analysed by GC-APCI-MS/MS. The pollutants soil burdens show a pronounced seasonality, a winter maximum for NPAHs and PAHs and a summer maximum for OPAHs. One order of magnitude more OPAH and parent PAH are found stored in forest soil than in nearby grassland soil. Among a number of 3-4 ring PAHs, the OPAHs benzanthrone and 6H-benzo(c,d)pyren-6-one, and the NPAHs 1- and 2-nitronaphthalene, 9-nitrophenanthrene and 7-nitrobenz(a)anthracene are found to re-volatilise from soils at a rural background site in central Europe in summer. At a receptor site in northern Europe, net deposition of polycyclic aromatic compounds (PACs) prevails and re-volatilisation occurs only sporadic. Re-volatilisation of a number of PACs, including strong mutagens, from soils in summer and even in winter indicates that long-range atmospheric transport of primary PAC emissions from central Europe to receptor areas might be enhanced by secondary emissions from soils.

Recent advances in carboxymethyl cellulose-based active and intelligent packaging materials: A comprehensive review.

Researchers have concentrated on innovative approaches to increase the shelf life of perishable food products and monitor their quality during storage and transportation as consumer demand for safe, environmentally friendly, and effective packaging develops. This comprehensive review aims to provide an overview of recent developments in carboxymethyl cellulose (CMC) chemical synthesis and its applications in active and intelligent packaging materials. It explores various methods for modifying cellulose to produce CMC and highlights the unique properties that make it suitable for addressing packaging industry challenges. The integration of CMC into active packaging systems, which helps reduce food waste and enhance food preservation, is discussed in depth. Furthermore, the integration of CMC in smart sensors and indicators for real-time monitoring and quality assurance in intelligent packaging is examined. The chemical synthesis of CMC and strategies to optimise its properties were studied, and the review concluded by examining the challenges and prospects of CMC-based packaging in the industry. This review is intended to serve as a valuable resource for researchers, industry professionals, and policymakers interested in the evolving landscape of CMC and its role in shaping the future of packaging materials.

2D Materials Beyond Post-AI Era: Smart Fibers, Soft Robotics, and Single Atom Catalysts.

Recent consecutive discoveries of various 2D materials have triggered significant scientific and technological interests owing to their exceptional material properties, originally stemming from 2D confined geometry. Ever-expanding library of 2D materials can provide ideal solutions to critical challenges facing in current technological trend of the fourth industrial revolution. Moreover, chemical modification of 2D materials to customize their physical/chemical properties can satisfy the broad spectrum of different specific requirements across diverse application areas. This review focuses on three particular emerging application areas of 2D materials: smart fibers, soft robotics, and single atom catalysts (SACs), which hold immense potentials for academic and technological advancements in the post-artificial intelligence (AI) era. Smart fibers showcase unconventional functionalities including healthcare/environmental monitoring, energy storage/harvesting, and antipathogenic protection in the forms of wearable fibers and textiles. Soft robotics aligns with future trend to overcome longstanding limitations of hard-material based mechanics by introducing soft actuators and sensors. SACs are widely useful in energy storage/conversion and environmental management, principally contributing to low carbon footprint for sustainable post-AI era. Significance and unique values of 2D materials in these emerging applications are highlighted, where the research group has devoted research efforts for more than a decade.

Impact of Ethanolic Thai Indigenous Leaf Extracts on Melanosis Prevention and Shelf-Life Extension of Refrigerated Pacific White Shrimp.

Shrimp has been known for its delicacy, but it undergoes rapid deterioration induced by biochemical and microbiological reactions. Melanosis is a major cause of discoloration associated with consumer rejection. All ethanolic extracts from different leaves including soursop, noni, and Jik leaves were dechlorophyllized via the "Green" sedimentation method before being used. The inhibitory activity against polyphenoloxidase (PPO) from Pacific white shrimp (Litopeneous vannamei) and the copper-chelating properties of varying extracts were compared. Soursop leaf extract (SLE) showed higher PPO inhibitory activity and copper-chelating ability than others (p < 0.05). Based on LC-MS, aempferol-3-O-rutinoside was identified as the most abundant compound, followed by catechin and neocholorigenic acid. The efficacy of SLE at different levels (0.25-1%) for inhibiting melanosis and preserving the quality of Pacific white shrimp was evaluated during refrigerated storage at 4 °C for 12 days in comparison with that of a 1.25% sodium metabisulfite (SMS)-treated sample. SLE at a level of 1% effectively retarded melanosis and bacterial growth, in which the total viable count did not exceed the microbial limit within 12 days. In addition, 1% SLE treatment impeded autolysis, reduced protein degradation and decomposition, and minimized lipid oxidation, as witnessed by the lower increases in pH, TVB-N, and TBARS values. Sensory evaluation indicated higher likeness scores and overall acceptability for SLE-1% and SMS-1.25% shrimps than those of the control and other samples. Therefore, SLE could be used as a natural alternative that effectively lowered the melanosis and quality loss of shrimp during refrigerated storage.

Direct Solution-Phase Synthesis and Functionalization of 2D WSe2 for Ambient Stability.

2H phase tungsten diselenide (WSe2 ) is a p-type 2D semiconductor from the transition metal dichalcogenides (TMDs) family with unique optoelectrical properties. Solution phase production of atomically thin WSe2 is challenging due to its instability under ambient conditions. We present a highly efficient and scalable solution method for simultaneously exfoliating and functionalizing WSe2 by leveraging the non-covalent interaction between mercapto-group and bulk WSe2 . Single and few-layer 2H phase pure WSe2 sheets of lateral size up to 5 µm with minimal basal plane defects, as revealed by XPS, Raman and FTIR spectroscopy, are produced in a water-ethanol mixture. Remarkably, WSe2 dispersion remains stable even at high concentrations (10 mg/mL) and exhibited high colloidal stability with a shelf-life exceeding a year. The findings from our study suggest that through precise manipulation of intercalation chemistry, mass production of solution-processable phase-sensitive 2D materials such as WSe2 can be achieved. This advancement holds great potential for facilitating their practical utilization in various real-world applications.

Gelatin/poly(vinyl alcohol)-based dual functional composite films integrated with metal-organic frameworks and anthocyanin for active and intelligent food packaging.

Innovative active and pH-colorimetric composite films were fabricated from gelatin/poly(vinyl alcohol) (Gel/PVA) integrated with copper-based metal-organic frameworks (Cu-MOFs) and red cabbage anthocyanin (RCA). The incorporation of Cu-MOFs improved the tensile strength, water resistance, and UV shielding properties of the developed composite films. The addition of anthocyanins and 3 wt% Cu-MOFs endowed the polymer matrix with excellent antioxidant (100 % against ABTS and DPPH radicals) and antibacterial (against Gram-positive and Gram-negative foodborne pathogenic bacteria) functions. The fabricated composite films exhibited significant color change at alkaline conditions of pH 7-12 and a marked color change upon exposure to ammonia. The designed indicator films used for shrimp freshness tracking and a visual color change from pink (for fresh shrimp) to green (for spoiled shrimp) was observed during storage at 28 °C for 24 h. The potential applications of the engineered composite films were studied by shrimp packaging, and the quality parameters of packaged samples were monitored during storage. The synergistic effects of adding anthocyanins and MOF nanostructures works for better product freshness preservation and responds well to shrimp spoilage level, introducing novel active and intelligent packaging options for practical smart packaging applications.

Trypsin from Pyloric Caeca of Asian Seabass: Purification, Characterization, and Its Use in the Hydrolysis of Acid-Soluble Collagen.

The study aimed to purify trypsin from the pyloric caeca of Asian seabass (Lates calcarifer), and investigate its proteolytic capability toward acid-soluble collagen (ASC) in comparison with commercial porcine trypsin (CPT). Trypsin was purified from pyloric caeca, a leftover from the evisceration process, via ammonium sulphate (40-60% saturation) precipitation, and a soybean trypsin inhibitor (SBTI)-Sepharose 4B column. A 18.5-fold purification and a yield of 15.2% were obtained. SDS-PAGE analysis confirmed a single band of trypsin with a molecular weight of 23.5 kDa. Purified trypsin also showed the single band in native-PAGE. The optimal pH and temperature of trypsin for BAPNA (the specific substrate for amidase) hydrolysis were 8.5 and 60 °C, respectively. The trypsin was stable within the pH range of 7.0-9.5 and temperature range of 25-55 °C. Protease inhibition study confirmed that the purified enzyme was trypsin. The purified trypsin had a Michaelis-Menten constant (Km) and catalytic constant (kcat) of 0.078 mM and 5.4 s-1, respectively, when BAPNA was used. For the hydrolysis of TAME (the specific substrate for esterase), the Km and Kcat were 0.09 mM and 4.8 s-1, respectively. Partially purified seabass trypsin (PPST) had a slightly lower hydrolysis capacity toward ASC than CPT, as evidenced by the lower degree of hydrolysis and protein degradation when the former was used. Both the α-chain and ß-chain became more degraded as the hydrolysis time increased. Based on MALDI-TOP, peptides with MW of 2992-2970 Da were dominant in the hydrolysates. Therefore, seabass trypsin could be used in the production of hydrolyzed collagen. It could have economic importance to the market, by replacing some commercial proteases, which have religious constraints.