Natural Polymer Encapsulated Zeolitic Imidazolate Framework-12 Composite toward Electrochemical Sensing of Antitumor Agent.

Encapsulation of natural polymer pectin (Pec) into a zeolitic imidazolate framework-12 (ZIF-12) matrix via a simple chemical method toward anticancer agent gallic acid (GA) detection is reported in this work. GA, a natural phenol found in many food sources, has gained attention by its biological effects on the human body, such as an antioxidant and anti-inflammatory. Therefore, it is crucial to accurately and rapidly determine the GA level in humans. The encapsulation of Pec inside the ZIF-12 has been successfully confirmed from the physiochemical studies such as XRD, Raman, FTIR, and XPS spectroscopy along with morphological FESEM, BET, and HRTEM characterization. Under optimized conditions, the Pec@ZIF-12 composite exhibits wide linear range of 20 nM-250 µM with a detection limit of 2.2 nM; also, it showed excellent selectivity, stability, and reproducibility. Furthermore, the real sample analysis of food samples including tea, coffee, grape, and pomegranate samples shows exceptional recovery percentage in an unspiked manner. So far, there is little literature for encapsulating proteins, enzymes, metals, etc., that have been reported; here, we successfully encapsulated a natural polymer Pec inside the ZIF-12 cage. This encapsulation significantly enhanced the composite electrochemical performance, which could be seen from the overall results. All of these strongly suggest that the proposed Pec@ZIF-12 composite could be used for miniaturized device fabrication for the evaluation of GA in both home and industrial applications.

3D Bioprinting of Sugar Beet Pectin through Horseradish Peroxidase-Catalyzed Cross-Linking.

Horseradish peroxidase (HRP)-mediated hydrogelation, caused by the cross-linking of phenolic groups in polymers in the presence of hydrogen peroxide (H2O2), is an effective route for bioink solidification in 3D bioprinting. Sugar beet pectin (SBP) naturally has cross-linkable phenols through the enzymatic reaction. Therefore, chemical modifications are not required, unlike the various polymers that have been used in the enzymatic cross-linking system. In this study, we report the application of SBP in extrusion-based bioprinting including HRP-mediated bioink solidification. In this system, H2O2 necessary for the solidification of inks is supplied in the gas phase. Cell-laden liver lobule-like constructs could be fabricated using bioinks consisting of 10 U/mL HRP, 4.0 and 6.0 w/v% SBP, and 6.0 × 106 cells/mL human hepatoblastoma (HepG2) cells exposed to air containing 16 ppm of H2O2 concurrently during printing and 10 min postprinting. The HepG2 cells enclosed in the printed constructs maintained their viability, metabolic activity, and hepatic functions from day 1 to day 7 of the culture, which indicates the cytocompatibility of this system. Taken together, this result demonstrates the potential of SBP and HRP cross-linking systems for 3D bioprinting, which can be applied in tissue engineering applications.

RG-I pectin-like polysaccharide from Rosa chinensis inhibits inflammation and fibrosis associated to HMGB1/TLR4/NF-κB signaling pathway to improve non-alcoholic steatohepatitis.

A novel RG-I pectin-like polysaccharide, YJ3A1, was purified from the flowers of Rosa chinensis and its structure and hepatoprotective effect in vivo and in vitro were investigated. The backbone of this polysaccharide is mainly composed of 1, 4-galactan, 1, 4-linked α-GalpA and 1, 2-linked α-Rhap disaccharide repeating unit attached by 1, 6-linked ß-Galp or 1, 5-linked α-Araf on C-4 of the Rhap. Interestingly, oral administration of YJ3A1 significantly ameliorates NASH-associated inflammation, oxidative stress and fibrosis and does not affect the liver morphology of normal mice at a dose of 50 mg/kg. The mechanism study suggests that the biological activity may associate to inactivating of high-mobility group box 1 protein (HMGB1)/TLR4/NF-κB and Akt signaling pathways by restraining the expression and release of HMGB1, thereby impeding the effect of NASH. The current findings outline a novel leading polysaccharide for new drug candidate development against NASH.

Design, fabrication, and evaluation of keratin and pectin incorporated supramolecular structured zero-oxidation state selenium nanogel blended 3D printed transdermal patch.

This study investigates the synthesis of selenium nanoparticles (SeNPs), owing to the low cost and abundance of selenium. However, the toxicity of SeNP prompts the development of a selenium nanocomposite (SeNC) containing pectin, keratin, and ferulic acid to improve the bioactivity of Se[0]. Further, incorporating the SeNC in a suitable formulation for drug delivery as a transdermal patch was worth studying. Accordingly, various analytical techniques were used to characterize the SeNPs and the SeNC, confirming successful synthesis and encapsulation. The SeNC exhibited notable particle size of 448.2 ± 50.2 nm, high encapsulation efficiency (98.90 % ± 2.4 %), 28.1 ± 0.45 drug loading, and sustained drug release at pH 5.5. Zeta potential and XPS confirmed the zero-oxidation state. The supramolecular structure was evident from spectral analysis endorsing the semi-crystalline nature of the SeNC and SEM images showcasing flower-shaped structures. Further, the SeNC demonstrated sustained drug release (approx. 22 % at 48 h) and wound-healing potential in L929 fibroblast cells. Subsequently, the SeNC loaded into a gelling agent exhibited shear thinning properties and improved drug release by nearly 58 %. A 3D printed reservoir-type transdermal patch was developed utilizing the SeNC-loaded gel, surpassing commercially available patches in characteristics such as % moisture uptake, tensile strength, and hydrophobicity. The patch, evaluated through permeation studies and CAM assay, exhibited controlled drug release and angiogenic properties for enhanced wound healing. The study concludes that this patch can serve as a smart dressing with tailored functionality for different wound stages, offering a promising novel drug delivery system for wound healing.

A methyl esterase from Bifidobacterium longum subsp. longum reshapes the prebiotic properties of apple pectin by triggering differential modulatory capacity in faecal cultures.

Pectin structures have received increasing attention as emergent prebiotics due to their capacity to promote beneficial intestinal bacteria. Yet the collective activity of gut bacterial communities to cooperatively metabolize structural variants of this substrate remains largely unknown. Herein, the characterization of a pectin methylesterase, BpeM, from Bifidobacterium longum subsp. longum, is reported. The purified enzyme was able to remove methyl groups from highly methoxylated apple pectin, and the mathematical modelling of its activity enabled to tightly control the reaction conditions to achieve predefined final degrees of methyl-esterification in the resultant pectin. Demethylated pectin, generated by BpeM, exhibited differential fermentation patterns by gut microbial communities in in vitro mixed faecal cultures, promoting a stronger increase of bacterial genera associated with beneficial effects including Lactobacillus, Bifidobacterium and Collinsella. Our findings demonstrate that controlled pectin demethylation by the action of a B. longum esterase selectively modifies its prebiotic fermentation pattern, producing substrates that promote targeted bacterial groups more efficiently. This opens new possibilities to exploit biotechnological applications of enzymes from gut commensals to programme prebiotic properties.

Enhancing Acid Resistance of Aspergillus niger Pectin Lyase through Surface Charge Design for Improved Application in Juice Clarification.

Pectin lyases (PNLs) can enhance juice clarity and flavor by degrading pectin in highly esterified fruits, but their inadequate acid resistance leads to rapid activity loss in juice. This study aimed to improve the acid resistance of Aspergillus niger PNL pelA through surface charge design. A modification platform was established by fusing pelA with a protein tag and expressing the fusion enzyme in Escherichia coli. Four single-point mutants were identified to increase the surface charge using computational tools. Moreover, the combined mutant M6 (S514D/S538E) exhibited 99.8% residual activity at pH 3.0. The M6 gene was then integrated into the A. niger genome using a multigene integration system to obtain the recombinant PNL AM6. Notably, AM6 improved the light transmittance of orange juice to 45.3%, which was 8.39 times higher than that of pelA. In conclusion, AM6 demonstrated the best-reported acid resistance, making it a promising candidate for industrial juice clarification.

Potential structure-function relationships of pectic polysaccharides from quinoa microgreens: Impact of various esterification degrees.

Pectic polysaccharides are one of the most vital functional ingredients in quinoa microgreens, which exhibit numerous health-promoting benefits. Nevertheless, the detailed information about the structure-function relationships of pectic polysaccharides from quinoa microgreens (QMP) remains unknown, thereby largely restricting their applications as functional foods or fortified ingredients. Therefore, to unveil the possible structure-function relationships of QMP, the mild alkali de-esterification was utilized to modify QMP, and then the correlations of esterification degrees of native and modified QMPs to their biological functions were systematically investigated. The results showed that the modified QMPs with different esterification degrees were successfully prepared by the mild alkali treatment, and the primary chemical structure (e.g., compositional monosaccharides and glycosidic linkages) of the native QMP was overall stable after the de-esterified modification. Furthermore, the results revealed that the antioxidant capacity, antiglycation effect, prebiotic potential, and immunostimulatory activity of the native QMP were negatively correlated to its esterification degree. In addition, both native and modified QMPs exerted immunostimulatory effects through activating the TLR4/NF-κB signaling pathway. These results are conducive to unveiling the precise structure-function relationships of QMP, and can also promote its applications as functional foods or fortified ingredients.

Structural analysis and in vitro fermentation characteristics of an Avicennia marina fruit RG-I pectin as a potential prebiotic.

Avicennia marina (Forssk.) Vierh. is a highly salt-tolerant mangrove, and its fruit has been traditionally used for treating constipation and dysentery. In this study, a pectin (AMFPs-0-1) was extracted and isolated from this fruit for the first time, its structure was analyzed, and the effects on the human gut microbiota were investigated. The results indicated that AMFPs-0-1 with a molecular weight of 798 kDa had a backbone consisting of alternating →2)-α-L-Rhap-(1→ and →4)-α-D-GalpA-(1→ residues and side chains composed of →3-α-L-Araf-(1→-linked arabinan with a terminal ß-L-Araf, →5-α-L-Araf-(1→-linked arabinan, and →4)-ß-D-Galp-(1→-linked galactan that linked to the C-4 positions of all α-L-Rhap residues in the backbone. It belongs to a type I rhamnogalacturonan (RG-I) pectin but has no arabinogalactosyl chains. AMFPs-0-1 could be consumed by human gut microbiota and increase the abundance of some beneficial bacteria, such as Bifidobacterium, Mitsuokella, and Megasphaera, which could help fight digestive disorders. These findings provide a structural basis for the potential application of A. marina fruit RG-I pectic polysaccharides in improving human intestinal health.

Effect of enzymatic modification on the structure and rheological properties of diluted alkali-soluble pectin fraction rich in RG-I.

This study focuses on pectin covalently linked in cell walls from two sources, apples and carrots, that was extracted using diluted alkali, and it describes changes in the rheological properties of diluted alkali-soluble pectin (DASP) due to enzymatic treatment. Given DASP's richness of rhamnogalacturonan I (RG-I), RG-I acetyl esterase (RGAE), rhamnogalacturonan endolyase (RGL), and arabinofuranosidase (ABF) were employed in various combinations for targeted degradation of RG-I pectin chains. Enzymatic degradations were followed by structural studies of pectin molecules using atomic force microscopy (AFM) as well as measurements of rheological and spectral properties. AFM imaging revealed a significant increase in the length of branched molecules after incubation with ABF, suggesting that arabinose side chains limit RG-I aggregation. Structural modifications were confirmed by changes in the intensity of bands in the pectin fingerprint and anomeric region on Fourier transform infrared spectra. ABF treatment led to a decrease in the stability of pectic gels, while the simultaneous use of ABF, RGAE, and RGL enzymes did not increase the degree of aggregation compared to the control sample. These findings suggest that the association of pectin chains within the DASP fraction may rely significantly on intermolecular interactions. Two mechanisms are proposed, which involve side chains as short-range attachment points or an extended linear homogalacturonan conformation favoring inter-chain interactions over self-association.

Ionic-physical-chemical triple cross-linked all-biomass-based aerogel for thermal insulation applications.

Aerogels, as a unique porous material, are expected to be used as insulation materials to solve the global environmental and energy crisis. Using chitosan, citric acid, pectin and phytic acid as raw materials, an all-biomass-based aerogel with high modulus was prepared by the triple strategy of ionic, physical and chemical cross-linking through directional freezing technique. Based on this three-dimensional network, the aerogel exhibited excellent compressive modulus (24.89 ± 1.76 MPa) over a wide temperature range and thermal insulation properties. In the presence of chitosan, citric acid and phytic acid, the aerogel obtained excellent fire safety (LOI value up to 31.2%) and antibacterial properties (antibacterial activity against Staphylococcus aureus and Escherichia coli reached 81.98% and 67.43%). In addition, the modified aerogel exhibited excellent hydrophobicity (hydrophobic angle of 146°) and oil-water separation properties. More importantly, the aerogel exhibited a biodegradation rate of up to 40.31% for 35 days due to its all-biomass nature. This work provides a green and sustainable strategy for the production of highly environmentally friendly thermal insulation materials with high strength, flame retardant, antibacterial and hydrophobic properties.

Physicochemical and antioxidant properties of pectin fractions extracted from lemon (Citrus Eureka) peels.

Pectin, a natural polysaccharide, holds versatile applications in food and pharmaceuticals. However, there is a need for further exploration into extracting novel functional fractions and characterizing them thoroughly. In this study, a sequential extraction approach was used to obtain three distinct lemon pectin (LP) fractions from lemon peels (Citrus Eureka): LP extracted with sodium acetate (LP-SA), LP extracted with ethylenediaminetetraacetic acid (LP-EDTA), and LP extracted with sodium carbonate and sodium borohydride (LP-SS). Comprehensive analysis revealed low methyl-esterification in all fractions. LP-SA and LP-SS displayed characteristics of rhamnogalacturonan-I type pectin, while LP-EDTA mainly consisted of homogalacturonan pectin. Notably, LP-SA formed self-aggregated particles with rough surfaces, LP-EDTA showed interlocking linear structures with smooth planes, and LP-SS exhibited branch chain structures with smooth surfaces. Bioactivity analysis indicated that LP-SA had significant apparent viscosity and ABTS radical scavenging activity, while both LP-EDTA and LP-SS showed excellent thermal stability according to thermogravimetric analysis (TGA). Furthermore, LP-SS exhibited remarkable gel-forming ability and significant hydroxyl free radicals scavenging activity. In conclusion, this study presents a novel method for extracting various lemon pectin fractions with unique structural and bioactive properties, contributing insights for advanced applications in the food and pharmaceutical sectors.

Role of acemannan and pectic polysaccharides in saline-water stress tolerance of Aloe vera (Aloe barbadensis Miller) plant.

This study investigates the impact of water and salinity stress on Aloe vera, focusing on the role of Aloe vera polysaccharides in mitigating these stresses. Pectins and acemannan were the most affected polymers. Low soil moisture and high salinity (NaCl 80 mM) increased pectic substances, altering rhamnogalacturonan type I in Aloe vera gel. Aloe vera pectins maintained a consistent 60 % methyl-esterification regardless of conditions. Interestingly, acemannan content rose with salinity, particularly under low moisture, accompanied by 90 to 150 % acetylation increase. These changes improved the functionality of Aloe vera polysaccharides: pectins increased cell wall reinforcement and interactions, while highly acetylated acemannan retained water for sustained plant functions. This study highlights the crucial role of Aloe vera polysaccharides in enhancing plant resilience to water and salinity stress, leading to improved functional properties.

Grafted calcium pectinate-whey protein isolate covalent immobilizers: Optimization, kinetics, thermodynamics, and application.

Whey protein isolate (WPI) was incorporated within calcium pectinate (CPT) beads in order to boost their anionic qualities and meliorate their glutaraldehyde (GA)-polyethyleneimine (PEI) grafting process. The Box-Behnken Design (BBD) verified that WPI inclusion significantly raised the GA-PEI-CPT-WPI beads immobilized ß-D-galactosidase (iß-GLD) activity. The BBD also revealed the optimal settings for WPI concentration, PEI pH, PEI concentration, and GA concentration, which were 2.91 %, 10.8, 3.5 %, and 2.24 %, respectively. The GA-PEI-CPT-WPI beads grafting process was scrutinized via FTIR, EDX, and SEM. The optimal GA-PEI-CPT-WPI immobilizers provided fine ß-GLD immobilization efficiencies, which reached up to 65.28 %. The free and GA-PEI-CPT-WPI iß-GLDs pH and temperature profiles were scrutinized. It was also unveiled that the thermal stability of the iß-GLD surpassed that of its free compeer as it provided lesser kd and ΔS values and larger t1/2, D-values, Ed, ΔH, and ΔG values. Furthermore, the iß-GLD provided 92.00±3.39 % activity after 42 storage days, which denoted its fine storage stability. The iß-GLD short duration (15 min) operational stability was also inspected, and 82.70±0.78 % activity was provided during the fifteenth degradation run. Moreover, the iß-GLD long duration (24 h) operational stability was inspected while degrading the lactose of buffered lactose solution (BLS) and cheese whey (CW). It was unveiled that 81.86±0.96 % and 73.58±2.24 % of the initial glucose were detected during the sixth degradation runs, respectively.

NKS1/ELMO4 is an integral protein of a pectin synthesis protein complex and maintains Golgi morphology and cell adhesion in Arabidopsis.

Adjacent plant cells are connected by specialized cell wall regions, called middle lamellae, which influence critical agricultural characteristics, including fruit ripening and organ abscission. Middle lamellae are enriched in pectin polysaccharides, specifically homogalacturonan (HG). Here, we identify a plant-specific Arabidopsis DUF1068 protein, called NKS1/ELMO4, that is required for middle lamellae integrity and cell adhesion. NKS1 localizes to the Golgi apparatus and loss of NKS1 results in changes to Golgi structure and function. The nks1 mutants also display HG deficient phenotypes, including reduced seedling growth, changes to cell wall composition, and tissue integrity defects. These phenotypes are comparable to qua1 and qua2 mutants, which are defective in HG biosynthesis. Notably, genetic interactions indicate that NKS1 and the QUAs work in a common pathway. Protein interaction analyses and modeling corroborate that they work together in a stable protein complex with other pectin-related proteins. We propose that NKS1 is an integral part of a large pectin synthesis protein complex and that proper function of this complex is important to support Golgi structure and function.

Antibacterial and Anti-Inflammatory Polysaccharide from Fructus Ligustri Lucidi Incorporated in PVA/Pectin Hydrogels Accelerate Wound Healing.

In cutaneous wound healing, an overproduction of inflammatory chemokines and bacterial infections impedes the process. Hydrogels can maintain a physiologically moist microenvironment, absorb chemokines, prevent bacterial infection, inhibit bacterial reproduction, and facilitate wound healing at a wound site. The development of hydrogels provides a novel treatment strategy for the entire wound repair process. Here, a series of Fructus Ligustri Lucidi polysaccharide extracts loaded with polyvinyl alcohol (PVA) and pectin hydrogels were successfully fabricated through the freeze-thaw method. A hydrogel containing a 1% mixing weight ratio of FLL-E (named PVA-P-FLL-E1) demonstrated excellent physicochemical properties such as swellability, water retention, degradability, porosity, 00drug release, transparency, and adhesive strength. Notably, this hydrogel exhibited minimal cytotoxicity. Moreover, the crosslinked hydrogel, PVA-P-FLL-E1, displayed multifunctional attributes, including significant antibacterial properties, earlier re-epithelialization, production of few inflammatory cells, the formation of collagen fibers, deposition of collagen I, and faster remodeling of the ECM. Consequently, the PVA-P-FLL-E1 hydrogel stands out as a promising wound dressing due to its superior formulation and enhanced healing effects in wound care.

The sequential microbial breakdown of pectin is the principal incident during water retting of jute (Corchorus spp.) bast fibres.

The extraction of bast fibres such as jute from plant stems involves the removal of pectin, hemicellulose, and other noncellulosic materials through a complex microbial community. A consortium of pectinolytic bacterial strains has been developed and commercialized to reduce the retting time and enhance fibre quality. However, there are currently no studies on jute that describe the structural changes and sequential microbial colonization and pectin loss that occur during microbe-assisted water retting. This study investigated the stages of microbial colonization, microbial interactions, and sequential degradation of pectic substances from jute bark under controlled and conventional water retting. The primary occurrence during water retting of bast fibres is the bacterially induced sequential breakdown of pectin surrounding the fibre bundles. The study also revealed that the pectin content of the jute stem significantly decreases during the retting process. These findings provide a strong foundation for improving microbial strains for improved pectinolysis with immense industrial significance, leading to a sustainable jute-based "green" economy.

Differences in physicochemical properties of pectin extracted from pomelo peel with different extraction techniques.

In order to obtain high yield pomelo peel pectin with better physicochemical properties, four pectin extraction methods, including hot acid extraction (HAE), microwave-assisted extraction (MAE), ultrasound-assisted extraction, and enzymatic assisted extraction (EAE) were compared. MAE led to the highest pectin yield (20.43%), and the lowest pectin recovery was found for EAE (11.94%). The physicochemical properties of pomelo peel pectin obtained by different methods were also significantly different. Pectin samples obtained by MAE had the highest methoxyl content (8.35%), galacturonic acid content (71.36%), and showed a higher apparent viscosity, thermal and emulsion stability. The pectin extracted by EAE showed the highest total phenolic content (12.86%) and lowest particle size (843.69 nm), showing higher DPPH and ABTS scavenging activities than other extract methods. The pectin extracted by HAE had the highest particle size (966.12 nm) and degree of esterification (55.67%). However, Fourier-transform infrared spectroscopy showed that no significant difference occurred among the different methods in the chemical structure of the extracted pectin. This study provides a theoretical basis for the industrial production of pomelo peel pectin.

Assessing high-temperature and pressure extraction of bioactive water-soluble polysaccharides from passion fruit mesocarp.

The mesocarp (albedo) of passion fruit is considered a waste product but rich in soluble fibers, especially pectins. Biological activity and health benefits of pectins have recently emerged, especially in colorectal cancer and attenuating inflammation. Pectin conventional extraction often uses mineral acids, which can be hazardous to the environment, and alternatives can be costly. Here, we assessed a high-temperature and pressure method to extract pectin from the passion fruit albedo and evaluated the differences from the water-soluble fractions extracted. HPSEC, HPAEC, FTIR-ATR, and HSQC-NMR were performed to identify and confirm the highly methylated homogalacturonan structures. The heat-modified samples showed a decreased molecular size compared to the untreated sample. Colorectal cancer cell lines showed reduced viability after being treated with different doses of modified samples, with two of them, LW-MP3 and 4, showing the most potent effects. All samples were detected inside cells by immunofluorescence assay. It was observed that LW-MP3 and 4 upregulated the p53 protein, indicating cell-cycle arrest and the cleaved caspase-9 in one of the cell lines, with LW-MP4 enhancing cell death by apoptosis. Since the modified samples were composed of hydrolyzed homogalacturonans, those probably were the responsible structures for these anti-cancer effects.

Ultrasound pretreatment enhances moisture migration and drying quality of mulberry via microstructure and cell-wall polysaccharides nanostructure modification.

The effects of ultrasound pretreatment (20 kHz, 30 W/L) on mulberries' texture, microstructure, characteristics of cell-wall polysaccharides, moisture migration, and drying quality were investigated over exposure times ranging from 15 to 45 min. Ultrasound induced softening of mulberry tissue, accompanied by an increase in water-soluble pectin and a decrease in chelate-soluble pectin and Na2CO3-soluble pectin concentrations. Noticeable depolymerization of the pectin nanostructure was observed in the pretreated mulberries, along with a decrease in molecular weight, attributed to side-chain structure cleavage. Ultrasound loosened the cell wall structure, increased free water content and freedom, thereby reducing water diffusion resistance. Ultrasound pretreatment reduced drying time by 11.2 % to 23.3 % at various processing times compared to controls. Due to significantly enhanced drying efficiency, the optimal pretreatment time (30 min) yielded dried mulberries with higher levels of total phenolics and total anthocyanins, along with an increased antioxidant capacity. The results of this study provide insights into the mechanisms by which ultrasound pretreatment can effectively enhance the mulberry drying process.

Near-freezing temperature suppresses avocado (Persea americana Mill.) fruit softening and chilling injury by maintaining cell wall and reactive oxygen species metabolism during storage.

To enhance the postharvest quality of avocado (Persea americana Mill.) fruit, this study investigates alterations in cell wall metabolism and reactive oxygen species (ROS) metabolism during near-freezing temperature (NFT) storage, and explores their impact on fruit softening. The fruit was stored at 25 °C, 5 °C, 2 °C, and NFT, respectively. NFT storage retarded firmness loss and chilling injury in comparison with 25 °C, 5 °C, and 2 °C. NFT storage delayed the decrease of ionic-soluble pectin (ISP) and cellulose (CLL) contents by suppressing cell wall degradation enzyme activities. Correlation analysis showed that cell wall degradation enzyme activities were positively correlated to rates of ethylene release and respiration. Moreover, NFT storage maintained higher levels of DPPH and ABTS scavenging abilities, activities of superoxide dismutase, peroxidase, and catalase, as well as ascorbate-glutathione cycle (ascorbic acid, glutathione, glutathione disulfide, ascorbate peroxidase, cycle-related enzymes), thereby inhibited the increase of ROS content, malondialdehyde content, and cell membrane permeability. Fruit firmness and chilling injury were correlated with the contents of hydrogen (H2O2), superoxide anion (O2.-), ISP, and CLL. These results suggested that NFT could suppress fruit softening and chilling injury by inhibiting cell wall degradation through delaying respiration and ethylene production and suppressing ROS production via activation of antioxidant systems, thereby maintaining quality and prolonged storage life during avocado fruit storage.