The regulation of postharvest strawberry quality mediated by abscisic acid under elevated CO2 stress.

Elevated CO2 was a potential strategy for strawberry preservation. However, the regulatory mechanism remained unclear. In current study, transcriptome analysis showed that elevated CO2 played important roles in regulating strawberry fruit quality at the transcriptional level, and plant hormones metabolism at least partially involved in the regulatory process. Further, ABA was demonstrated to play important roles in the response to elevated CO2. Elevated CO2 inhibited the accumulation of ABA, which was 61% lower than that in control. Elevated CO2 repressed ABA synthesis by inhibiting NCED activity and the expression of FaNCED1/2, leading to the reduction of ABA accumulation as a result. Meanwhile, elevated CO2 also decreased ABA sensitivity by down-regulating FaSnRK2.4/2.6 and FaABI5 expression. The dual down-regulation of ABA signaling accounted for the regulation of fruit quality under elevated CO2 treatment. These results provide new insights into the mechanism of strawberry fruit response to elevated CO2.

The combination of metabolome and transcriptome clarifies the inhibition of the Alternaria toxin accumulation by methyl ferulate.

As one of the most typical pathogens in fruit postharvest diseases, Alternaria alternata (A. alternata) can produce Alternaria toxins (ATs) aggravating fruit decay and harming human health. In this study, ATs (tenuazonic acid, alternariol monomethyl ether, and alternariol) production was inhibited effectively by 200 and 8000 mg/L MF (methyl ferulate) in vitro and in vivo. 1-Octen-3-ol and 3-octanol were the potential iconic volatile organic compounds of ATs (R2 > 0.99). MF induced oxidative stress, resulting in physiological and metabolic disorders, membrane lipid oxidation and cell damage. It decreased precursors and energy supply by disturbing amino acid metabolism, ABC transporters, citrate cycle, pentose and glucuronate interconversions to regulate ATs synthesis. MF down-regulated the genes related to ATs synthesis (PksJ, AaTAS1, and OmtI), transport (AaMFS1 and MFS), and pathogenicity to affect ATs production and virulence. This study provided a theoretical basis for the control of ATs production.

Curdlan-polyphenol complexes prepared by pH-driven effectively enhanced their physicochemical stability, antioxidant and prebiotic activities.

In this study, the curdlan-polyphenol complexes were constructed by a pH-driven method. The interaction between curdlan and various hydrophobic polyphenols (curcumin, quercetin, and chlorogenic acid) was investigated. Curdlan could self-assemble into particles for loading polyphenols through hydrogen bonding and hydrophobic interactions. The three polyphenols were embedded in curdlan in an amorphous state. The curdlan-curcumin complex showed the lowest viscoelasticity but exhibited the highest curcumin loading ability (34.04 ± 1.73 mg/g). However, the curdlan-chlorogenic acid complex emerged the opposite trend, indicating that the loading capacity was associated with the hydrophobicity of polyphenols. The antioxidant activity of curdlan significantly increased after combining with polyphenols, which could be maintained during in vitro simulated gastrointestinal digestion. In particular, the curdlan-quercetin complex exhibited the highest antioxidant activity and short-chain fatty acid concentration, which could influence gut microbiota composition by promoting the proliferation of Prevotella and inhibiting the growth of Escherichia_Shigella. In conclusion, the curdlan-polyphenol complexes prepared by an alcohol-free pH-driven method could effectively enhance the gastrointestinal stability of polyphenols as well as increase the antioxidant and prebiotic activities of curdlan, which could be applied as a functional ingredient to improve gut health.

1-MCP treatment improves the postharvest quality of Jinxiu yellow peach by regulating cuticular wax composition and gene expression during cold storage.

This study aimed to analyze the effect of 1-methylcyclopropene (1-MCP) treatment on the postharvest quality, epidermal wax morphology, composition, and gene expression of Jinxiu yellow peach during cold storage. The results showed that 1-MCP treatment could maintain the postharvest quality of peach fruit as compared to control (CK) during cold storage. The wax crystals of peach fruit were better retained by 1-MCP, and they still existed in 0.6 and 0.9 µL/L 1-MCP treated fruit at 36 days. The total wax content in all the fruit increased first and then decreased during cold storage. Meanwhile, n-alkanes and primary alcohols were the main wax components. Compared to CK, 1-MCP treatment could delay the reduction of wax content during cold storage. The correlation analysis indicated that the postharvest quality of yellow peach was mainly affected by the contents of fatty acids and triterpenoids in cuticular wax. The transcriptomics results revealed PpaCER1, PpaKCS, PpaKCR1, PpaCYP86B1, PpaFAR, PpaSS2, and PpaSQE1 played the important roles in the formation of peach fruit wax. 1-MCP treatment upregulated PpaCER1 (18785414, 18786441, and 18787644), PpaKCS (18774919, 18789438, and 18793503), PpaKCR1 (18790432), and PpaCYP86B1 (18789815) to deposit more n-alkanes and fatty acids during cold storage. This study could provide a new perspective for regulating the postharvest quality of yellow peach in view of the application of cuticular wax. PRACTICAL APPLICATION: 'Jinxiu' yellow peach fruit is favorable among consumers because of its high commercial value. However, it ripens and deteriorates rapidly during storage, leading to serious economic loss and consumer disappointment. The effect of 1-methylcyclopropene (1-MCP) treatment on the postharvest quality, epidermal wax morphology, composition, and genes regulation of 'Jinxiu' yellow peach during cold storage was assessed. Compared to control, 1-MCP treatment could retain the storage quality of yellow peach by affecting cuticular wax composition and gene expression. This study could provide new perspective for regulating the postharvest quality of yellow peach in view of the application of cuticular wax.

Application of L-Cysteine Hydrochloride Delays the Ripening of Harvested Tomato Fruit.

Fruit ripening is controlled by internal factors such as hormones and genetic regulators, as well as external environmental factors. However, the impact of redox regulation on fruit ripening remains elusive. Here, we explored the effects of L-cysteine hydrochloride (LCH), an antioxidant, on tomato fruit ripening and elucidated the underlying mechanism. The application of LCH effectively delayed tomato fruit ripening, leading to the suppression of carotenoid and lycopene biosynthesis and chlorophyll degradation, and a delayed respiration peak. Moreover, LCH-treated fruit exhibited reduced hydrogen peroxide (H2O2) accumulation and increased activities of superoxide dismutase (SOD), catalase (CAT), and monodehydroascorbate reductase (MDHAR), compared with control fruit. Furthermore, transcriptome analysis revealed that a substantial number of genes related to ethylene biosynthesis (ACS2, ACS4, ACO1, ACO3), carotenoid biosynthesis (PSY, PDS, ZDS, CRTISO), cell wall degradation (PG1/2, PL, TBG4, XTH4), and ripening-related regulators (RIN, NOR, AP2a, DML2) were downregulated by LCH, resulting in delayed ripening. These findings suggest that the application of LCH delays the ripening of harvested tomato fruit by modulating the redox balance and suppressing the expression of ripening-related genes.

A novel edible solid fat substitute: Preparation of biphasic stabilized bigels based on glyceryl monolaurate and gellan gum.

Solid fats contribute to a delicate and pleasant flavor for food, but its excessive intake increases the risk of cardiovascular disease. Bigel is considered a promising solid fat substitute as it significantly reduces fat content while meeting consumer demands for food flavor and a balanced diet. In this study, bigels were prepared by mixing glyceryl monolaurate-based oleogel (10 wt%) and gellan gum-based hydrogel (0.8 wt%) at ratios of 1:3, 1:1, and 3:1. The microscopic results indicated that the oleogel/hydrogel ratios influenced the structure of bigels, forming oil-in-water, bi-continuous, and water-in-oil bigels with the increase of oleogel proportion, respectively. All bigels presented a semi-solid structure dominated by elasticity, and their hardness, gumminess, chewiness, and cohesiveness increased with the enhancement of hydrogel proportion. Among them, the bigels (S25:L75 and S25:H75) prepared with an oleogel/hydrogel ratio of 1:3 showed excellent freeze-thaw stability, maintaining an oil holding capacity of >95 % after three freeze-thaw cycles. Meanwhile, they also presented good oxidative stabilities, where the peroxide values and malondialdehyde contents were below 0.07 g/100 g and 1.5 mg MDA/kg at 12 d, respectively. Therefore, S25:L75 and S25:H75 are expected to be green, low-cost, healthy, and sustainable alternatives to solid fats.

Grafting chlorogenic acid enhanced the antioxidant activity of curdlan oligosaccharides and modulated gut microbiota.

In this study, the effects of grafting chlorogenic acid (CA) on the antioxidant and probiotic activities of curdlan oligosaccharides (CDOS) were investigated. CDOS with degrees of polymerization of 3-6 was first obtained by degradation of curdlan with hydrogen peroxide and then grafted with CA using a free radical-mediated method under an ultrasonication-assisted Fenton system. The thermal stability and antioxidant ability of CDOS were enhanced after grafting with CA. In vitro fermentation, supplementation of CDOS-CA stimulated the proliferation of Prevotella and Faecalibacterium while inhibiting the growth of harmful microbiota. Notably, the concentration of total short-chain fatty acids and the relative abundance of beneficial bacteria markedly increased after fermentation of CDOS-CA, indicating that CA grafting could improve the probiotic activity of CDOS. Overall, the covalent binding of CDOS and CA could enhance the antioxidant and probiotic activities of CDOS, suggesting potential improvements in gastrointestinal and colonic health.

Development of a chitosan/pectin-based multi-active food packaging with both UV and microbial defense functions for effectively preserving of strawberry.

Multi-active food packaging was prepared for strawberry fruit preservation where epigallocatechin gallate (EGCG)-containing pectin matrix and natamycin (NATA)-containing chitosan (CS) matrix were utilized to complete LBL electrostatic self-assembly. The results showed that the physicochemical properties of the multi-active packaging were closely related to the addition of NATA and EGCG. It was found that NATA and EGCG were embedded in the CS/pectin matrix through intermolecular hydrogen bonding interactions. The CN/PE 15 % multi-active films prepared based on the spectral stacking theory formed a barrier to UV light in the outer layer, exhibited excellent NATA protection under UV light exposure conditions at different times, and provided long-lasting and sustained bacterial inhibition in the inner layer. In addition, the CN/PE 15 % multi-active packaging extended the shelf life of strawberry at room temperature compared with the control samples. In conclusion, the developed CN/PE 15 % packaging provided potential applications for multi-active food packaging materials.

How to effectively and greenly prepare multi-scale structural starch nanoparticles for strengthening gelatin film (ultrasound-Fenton system).

Ultrasound (US) assisted with Fenton (US-Fenton) reaction was developed to efficiently and greenly prepare starch nanoparticles (SNPs) that were employed as nanofillers to enhance gelatin (G) film properties. Compared to Fenton reaction alone, US-Fenton reaction significantly improved preparation efficiency and dispersion of SNPs (p < 0.05). An optimal US-Fenton reaction parameter (300 mM H2O2, ascorbic acid 55 mM, US 45 min) was found to prepare SNPs with uniform sizes (50-90 nm) and low molecular weight (Mn 7.91 × 105 Da). The XRD, FT-IR, and SAXS analysis revealed that the US-Fenton reaction degraded the amorphous and crystalline zones of starch from top to down, leading to the collapse of the original layered structure starch and the progressive formation of SNPs. The different sizes of SNPs were selected to prepare the composite films. The G-SNP3 film (with 50-90 nm SNPs) showed the most outstanding UV blocking, tensile, and barrier properties. Especially, the tensile strength of G-5%SNP3 film (containing 5 % SNPs) increased by 156 % and 6 % over that of G film and G-5%SNP2 film (containing 5%SNPs with 100-180 nm), respectively. Therefore, the nanomaterial was promisingly prepared by the US-Fenton system and provided a strategy for designing and producing nanocomposite films.

Ultrasonication significantly enhances grafting efficiency of chitosan-ferulic acid conjugate and improves its film properties under Fenton system.

Ultrasonication (US)-assisted Fenton-system (US-Fenton) with different US time was developed for synthesizing chitosan (CS)-ferulic acid (FA) conjugates. The optimal US-Fenton for a suitable time was selected for preparing a film with CS-FA conjugate and its structural, functional, rheological, and physical properties were also investigated. Compared with Fenton-system, US-Fenton enhanced the grafting ratio of the conjugates, which increased firstly and then decreased as US time. The conjugate obtained by US-Fenton for 1 min (FUS1) possessed the highest grafting ratio (121.28 mg FA/g) and its grafting time was also shortened from 12 h to 1 min contrasted with Fenton grafted method. Structural characterization results showed that FA was conjugated on CS via ester and amide bonds with decreased crystallinity. Scanning electron microscopy and molecular weight analysis indicated that the degradation degree of CS-FA conjugates increased with US time. The DPPH and ABTS radical-scavenging activities of FUS1 were the closest to ascorbic acid, and it also showed the best antibacterial effect among the test conjugates. Accordingly, FUS1 was selected to obtain the film for contrasting with CS film. FUS1 film solution exhibited a decreased viscosity. In comparison to CS film, UV transmittance of FUS1 film approached zero, and its moisture, oxygen, and carbon dioxide permeabilities significantly decreased (P < 0.05). Moreover, its water solubility and tensile strength increased by 58.09% and 25.72% than those of CS film, respectively. Therefore, US-Fenton for 1 min could be a promising method for efficiently preparing active food package materials and FUS1 film possessed broad application prospects.

Transcriptomics and gas chromatography-mass spectrometry metabolomics reveal the mechanism of heat shock combined with 1-methylcyclopropene to regulate the cuticle wax of jujube fruit during storage.

Cuticle wax is closely related to fruit quality during storage. In this study, changes in epidermal wax morphology, composition, and genes regulation induced by heat shock (HT), 1-methylcyclopropene (1-MCP) or their combination (HT + 1-MCP) were investigated in jujube fruit during cold storage. HT, 1-MCP, or HT + 1-MCP caused a smoother wax layer and fewer micro-cracks compared to the control (CK) during cold storage. It was confirmed that acids and terpenoids were the main wax components by gas chromatography-mass spectrometry. HT + 1-MCP and 1-MCP treatments could significantly increase (p < 0.05) the wax content at 45 d of cold storage. The transcriptomics results indicated that HT + 1-MCP treatment up-regulated FATB, FATB, FAB2, FAD2 and CYP716A, and maintained the wax content of jujube fruit during cold storage. These results could provide new perspective for regulating the cuticle characteristics to extend the shelf life of jujube fruit.

Fabrication and Characterization of Eco-Friendly Polyelectrolyte Bilayer Films Based on Chitosan and Different Types of Edible Citrus Pectin.

The eco-friendly polyelectrolyte bilayer films were prepared by layer-by-layer (LBL) casting method using chitosan (CS) and four types of edible citrus pectin as film substrates. The results showed that the polyelectrolyte bilayer films exhibited excellent comprehensive properties. Furthermore, the interaction between CS and pectin was closely related to the degree of methyl-esterification (DM), molecular weight (Mw), and zeta potential of pectin. The low DM, Mw, and high zeta potential of the low methyl-esterified pectin (LM) resulted in a denser internal structure of the bilayer film, stronger UV shielding performance, and stronger gas barrier ability. The high DM and Mw of the high methyl-esterified pectin (HM) endow the bilayer film with stronger mechanical properties, thermal stability, and antifogging property. The microstructural and spectroscopic analysis showed that there are hydrogen bonds and electrostatic interactions between the layers. Overall, the developed CS-pectin polyelectrolyte bilayer films provided potential applications for food bioactive packaging.

Gas Chromatography-Mass Spectrometry Metabolite Analysis Combined with Transcriptomic and Proteomic Provide New Insights into Revealing Cuticle Formation during Pepper Development.

The cuticle plays an important role for the quality of pepper fruit. However, the molecular mechanism of cuticle formation in pepper fruit remains unclear. Our results showed that the wax was continuously accumulated during pepper development, while the cutin monomer first increased and then decreased. Hexadecanoic acid and 10,16-hydroxyhexadecanoic acid were the main components of wax and cutin, respectively. Combined with transcriptome and proteome, the formation patterns of wax and cutin polyester network for pepper cuticle was proposed. The 18 pairs of consistent expression genes and proteins involved in cuticle formation were revealed. Meanwhile, 12 key genes were screened from fatty acid biosynthesis, biosynthesis of unsaturated fatty acids, fatty acid elongation, cutin, suberine, and wax biosynthesis, glycerolipid metabolism, and transport pathway. This study would provide important candidate genes and theoretical basis for the molecular mechanism of cuticle formation, which is essential for the breeding of peppers.

Fabrication and characterization of pullulan-based composite films incorporated with bacterial cellulose and ferulic acid.

Pullulan-based composite films incorporated with bacterial cellulose (BC) and ferulic acid (FA) were prepared by solution casting method. The rheological, morphological, barrier, optical, anti-fogging, and antioxidant properties of pullulan-based composite films doped with BC and FA were investigated. The rheological results showed that all film-forming solution was pseudoplastic fluid and its viscosity increased with the increase of BC content. An appropriate BC (2 %) and FA were uniformly dispersed in pullulan to form uniform and dense composite films. With the increase of BC content, the roughness and opacity of composite films increased while their UV-vis barrier performance was improved by incorporating BC and FA. Fourier transform infrared spectrometer analysis demonstrated that hydrogen bond interactions among pullulan, BC, and FA were found, and incorporating BC could increase the crystallinity of the composite films, thus enhancing their mechanical, barrier, hydrophobic, and thermal stability properties. Pullulan-based composite films incorporated with 2 % BC and FA (P-BC2-FA) showed better mechanical properties, water, oxygen, and carbon dioxide barrier performances, and its water contact angle value also increased compared with control, respectively. P-BC2-FA film showed superior anti-fogging and antioxidant activities. These results indicate that the P-BC2-FA film are expected to be a potential target of bioactive packaging.

Substrates and Loaded Iron Ions Relative Position Influence the Catalytic Characteristics of the Metalloenzymes Angelica archangelica Flavone Synthase I and Camellia sinensis Flavonol Synthase.

Metalloenzymes are a class of enzymes that catalyze through the metal ions they load. Angelica archangelica flavone synthase I (AnFNS I) and Camellia sinensis flavonol synthase (CaFLS), both of which belong to metalloenzymes, have highly similar structures and metal catalytic cores. However, these two enzymes catalyze the same substrate to produce significantly different products. To identify the cause for the differences in the catalytic characteristics of AnFNS I and CaFLS, their protein models were constructed using homology modeling. Structural alignment and molecular docking was also used to elucidate the molecular basis of the differences observed. To analyze and verify the cause for the differences in the catalytic characteristics of AnFNS I and CaFLS, partial fragments of AnFNS I were used to replace the corresponding fragments on CaFLS, and the catalytic characteristics of the mutants were determined by bioconversion assay in E. coli and in vitro catalytic test. The results suggest that the difference in catalytic characteristics between AnFNS I and CaFLS is caused by the depth of the active pockets and the relative position of the substrate. Mutant 10 which present similar dock result with AnFNS I increased the proportion of diosmetin (a flavone) from 2.54 to 16.68% and decreased the proportion of 4'-O-methyl taxifolin (a flavanol) from 47.28 to 2.88%. It was also indicated that the atoms in the substrate molecule that determine the catalytic outcome may be H-2 and H-3, rather than C-2 and C-3. Moreover, it is speculated that the change in the catalytic characteristics at the changes relative spatial position of H-2/H-3 of hesperetin and the loaded carbonyl iron, caused by charged residues at the entrance of the active pocket, is the key factor for the biosynthesis of flavone from flavanone.

High pressure processing improves the texture quality of fermented minced pepper by maintaining pectin characteristics during storage.

Texture quality affects the sensory and market acceptance of fermentation minced pepper (FMP), but it will deteriorate during storage. Thus, high pressure processing (HPP) and thermal pasteurization (TP) were used to improve the texture quality of FMP during storage. The results showed that variations in texture quality and pectin characteristics under HPP and TP treatments were similar during storage. The hardness, cell wall material (CWM) and sodium carbonate-soluble pectin (SSP) content, water-soluble pectin (WSP) molecular weight (Mw ) decreased, while WSP content and sodium chelate-soluble pectin (CSP) Mw increased after storage. HPP-treated FMP showed higher hardness (66.64-85.95 N) than that in TP-treated one (57.23-62.72 N) during storage. Rhamnose (Rha), arabinose, mannose, and glucose were the crucial compositions in three pectins, and their total molar ratios, respectively, reached 89.19% and 87.97% after HPP and TP treatment. However, the molar ratio of most monosaccharide in three pectins decreased after storage. Atomic force microscope images indicated the short chains and branch structures increased but aggregates decreased in most pectin components during storage. Pearson correlation analysis demonstrated FMP hardness was extremely (p < 0.01) positively correlated with CWM and SSP content, and extremely (p < 0.01) negatively correlated with WSP content. Compared to TP treatment, HPP presented higher hardness, SSP content and Mw , Rha content, CSP Mw , and lower WSP content during storage. Hence, HPP was an effective method to improve the texture quality of FMP by maintaining pectin characteristics during storage. PRACTICAL APPLICATION: Softening is one of the main factors affecting market value and consumer preferences for FMP, and it is closely related to the modification and depolymerization of pectin. Changes of texture quality and pectin properties in HPP- and TP-treated FMP during storage were assessed, including hardness, the content, monosaccharide compositions, Mw distribution, and nanostructure of WSP, SSP, and CSP. Compared to TP treatment, HPP could effectively improve the texture quality of FMP by inhibiting pectin degradation during storage. All the findings presented in this study would help to provide new insights into regulating the texture quality of FMP.

Zein/pullulan complex colloidal particle-stabilized Pickering emulsions for oral delivery of polymethoxylated flavones: protection effect and in vitro digestion.

BACKGROUND: Polymethoxylated flavones (PMFs) show multiple biological functions, while their high hydrophobicity leads to a low bioaccessibility and limits their wide applications. The design of a reasonable food-grade drug delivery system is an effective strategy to improve the low bioaccessibility of PMFs. In this study, sinensetin, tangeretin and nobiletin were encapsulated in Pickering emulsions stabilized by zein/pullulan complex colloidal particles (ZPPs), and the protection effect and in vitro digestion were characterized. RESULTS: Rheological analysis revealed that ZPP-Pickering emulsion loading with PMFs maintained a strong gel-like network structure. Moreover, the ability to scavenge free radicals of PMFs was improved by the emulsion delivery system. The antioxidant activity of PMFs encapsulated in Pickering emulsion was positively correlated with the oil volume fraction (φ). ZPP-Pickering emulsion loading with PMFs can effectively delay lipid oxidation, and the φ (70%) of Pickering emulsion showed the most pronounced effects, in which the lipid hydroperoxide content and malondialdehyde content decreased by 64.3% and 38.3% after 15 days of storage, compared with the bulk oil group, respectively. The bioaccessibility of the three PMFs has been increased by ZPP-Pickering emulsion simultaneously and it presented the highest values as its φ was 50%, in which the bioaccessibility of sinensetin, tangeretin and nobiletin increased by 2.5, 3.2 and 3.9 times, compared with the bulk oil group, respectively. CONCLUSION: Pickering emulsion stabilized by ZPPs is an excellent nutrient delivery system for delivering three PMFs simultaneously and imparting functional properties to bioactive delivery systems. © 2022 Society of Chemical Industry.

Nobiletin, a hexamethoxyflavonoid from citrus pomace, attenuates G1 cell cycle arrest and apoptosis in hypoxia-induced human trophoblast cells of JEG-3 and BeWo via regulating the p53 signaling pathway.

BACKGROUND: Hypoxia is associated with abnormal cell apoptosis in trophoblast cells, which causes fetal growth restriction and related placental pathologies. Few effective methods for the prevention and treatment of placenta-related diseases exist. Natural products and functional foods have always been a rich source of potential anti-apoptotic drugs. Nobiletin (NOB), a hexamethoxyflavonoid derived from the citrus pomace, shows an anti-apoptotic activity, which is a non-toxic constituent of dietary phytochemicals approved by the Food and Drug Administration. However, their effects on hypoxia-induced human trophoblast cells have not been fully studied. OBJECTIVE: The aim of this study was to investigate the protective effects of NOB on hypoxia-induced apoptosis of human trophoblast JEG-3 and BeWo cells, and their underlying mechanisms. DESIGN: First, the protective effect of NOB on hypoxia-induced apoptosis of JEG-3 and BeWo cells was studied. Cell viability and membrane integrity were determined by CCK-8 assay and lactate dehydrogenase activity, respectively. Real Time Quantitative PCR (RT-qPCR) and Western blot analysis were used to detect the mRNA and protein levels of HIF1α. Propidium iodide (PI)-labeled flow cytometry was used to detect cell cycle distribution. Cell apoptosis was detected by flow cytometry with Annexin V-FITC and PI double staining, and the expression of apoptosis marker protein cl-PARP was detected by Western blot analysis. Then, the molecular mechanism of NOB against apoptosis was investigated. Computer molecular docking and dynamics were used to simulate the interaction between NOB and p53 protein, and this interaction was verified in vitro by Ultraviolet and visible spectrum (UV-visible spectroscopy), fluorescence spectroscopy and circular dichroism. Furthermore, the changes in the expression of p53 signaling pathway genes and proteins were detected by RT-qPCR and Western blot analysis, respectively. RESULTS: Hypoxia treatment resulted in a decreased cell viability and cell membrane integrity in JEG-3 and BeWo cell lines, and an increased expression of HIF1α, cell cycle arrest in the G1 phase, and massive cell apoptosis, which were alleviated after NOB treatment. Molecular docking and dynamics simulations found that NOB spontaneously bonded to human p53 protein, leading to the change of protein conformation. The intermolecular interaction between NOB and human p53 protein was further confirmed by UV-visible spectroscopy, fluorescence spectroscopy and circular dichroism. After the treatment of 100 µM NOB, a down-regulation of mRNA and protein levels of p53 and p21 and an up-regulation of BCL2/BAX mRNA and protein ratio were observed in JEG-3 cells; however, there was also a down-regulation of mRNA and protein levels observed for p53 and p21 in BeWo cells after the treatment of NOB. The BCL2/BAX ratio of BeWo cells did not change after the treatment of 100 µM NOB. CONCLUSION: NOB attenuated hypoxia-induced apoptosis in JEG-3 and BeWo cell lines and might be a potential functional ingredient to prevent pregnancy-related diseases caused by hypoxia-induced apoptosis. These findings would also suggest the exploration and utilization of citrus resources, and the development of citrus industry.

Effect of hydrothermal-calcium chloride treatment on pectin characteristics and related quality in green peppers during storage.

Effects of hydrothermal (HT)-calcium chloride (CaCl2) treatment on pectin characteristics and related quality in green peppers during storage were assessed. The results showed that the changes of physicochemical quality in all green peppers were similar during storage. Weight loss percentage increased, firmness, the content of free water and bound water decreased during storage. Water-soluble pectin (WSP) notably increased, but sodium carbonate-soluble pectin (SSP) and chelate-soluble pectin (CSP) decreased. Galacturonic acid (GalUA), rhamnose (Rha), galactose (Gal), and arabinose (Ara) were the crucial compositions in the backbone and branched chains of pectin in green peppers. Rha and Gal increased, but Ara decreased in pectin after storage. The changes in the ratio of Rha/GalUA, Ara/Gal, and (Gal + Ara)/Rha represented that the backbone and branched chains of pectin in green peppers depolymerized to some extent after storage. Comparing with other green peppers, HT-CaCl2 treated green peppers posed lower weight loss percentage and WSP content, higher firmness, the content of free water, bound water, SSP, and CSP during storage. Otherwise, most pectin compositions in HT-CaCl2 treated green peppers showed high molar ratio after storage. Hence, HT-CaCl2 treatment was an effective way to retain pectin characteristics and related quality of green peppers, and further inhibited the softening of green peppers during storage.

Screening and heterologous expression of flavone synthase and flavonol synthase to catalyze hesperetin to diosmetin.

OBJECTIVES: In this study, 44 flavone synthases (FNS) and flavonol synthases (FLS) from different origins were collected. The instability index and conserved domain of the enzymes were analyzed through bioinformatics analysis, the results of which allowed us to screen suitable enzymes for constructing recombinant Escherichia coli. Defective enzymes were selected as controls. RESULTS: Native- and sodium dodecyl sulfate-polyacrylamide gel electrophoresis were conducted to isolate the heterologously expressed proteins. Liquid chromatography-mass spectrometry, 1H nuclear magnetic resonance, and ultra-performance liquid chromatography were performed to qualitatively and quantitatively analyze the products. The cellular transformation results showed that recombinant E. coli catalyzed the synthesis of diosmetin from hesperetin, and in vitro catalysis showed that heterologously expressed FNS/FLS played a catalytic role in this reaction. AnFNS (from Angelica archangelica) showed the highest substrate conversion (38.80% for cellular transformation, 12.93% for in vitro catalysis). CONCLUSIONS: The catalytic capacity of FNS/FLS from different origins exhibited the expected results, indicating that bioinformatics analysis is useful for screening enzymes. In addition, the catalytic properties of AnFNS and CaFLS (from Camellia sinensis) differed significantly, although these enzymes are structurally similar. Based on this difference, C-2 was predicted as the key site for FNS/FLS catalytic synthesis of diosmetin rather than C-3.