A flavonoid metabolon: cytochrome b5 enhances B-ring trihydroxylated flavan-3-ols synthesis in tea plants.

Flavan-3-ols are prominent phenolic compounds found abundantly in the young leaves of tea plants. The enzymes involved in flavan-3-ol biosynthesis in tea plants have been extensively investigated. However, the localization and associations of these numerous functional enzymes within cells have been largely neglected. In this study, we aimed to investigate the synthesis of flavan-3-ols in tea plants, particularly focusing on epigallocatechin gallate. Our analysis involving the DESI-MSI method to reveal a distinct distribution pattern of B-ring trihydroxylated flavonoids, primarily concentrated in the outer layer of buds. Subcellular localization showed that CsC4H, CsF3'H, and CsF3'5'H localizes endoplasmic reticulum. Protein-protein interaction studies demonstrated direct associations between CsC4H, CsF3'H, and cytoplasmic enzymes (CHS, CHI, F3H, DFR, FLS, and ANR), highlighting their interactions within the biosynthetic pathway. Notably, CsF3'5'H, the enzyme for B-ring trihydroxylation, did not directly interact with other enzymes. We identified cytochrome b5 isoform C serving as an essential redox partner, ensuring the proper functioning of CsF3'5'H. Our findings suggest the existence of distinct modules governing the synthesis of different B-ring hydroxylation compounds. This study provides valuable insights into the mechanisms underlying flavonoid diversity and efficient synthesis and enhances our understanding of the substantial accumulation of B-ring trihydroxylated flavan-3-ols in tea plants.

The Anti-Inflammatory and Curative Exponent of Probiotics: A Comprehensive and Authentic Ingredient for the Sustained Functioning of Major Human Organs.

Several billion microorganisms reside in the gastrointestinal lumen, including viruses, bacteria, fungi, and yeast. Among them, probiotics were primarily used to cure digestive disorders such as intestinal infections and diarrhea; however, with a paradigm shift towards alleviating health through food, their importance is large. Moreover, recent studies have changed the perspective that probiotics prevent numerous ailments in the major organs. Probiotics primarily produce biologically active compounds targeting discommodious pathogens. This review demonstrates the implications of using probiotics from different genres to prevent and alleviate ailments in the primary human organs. The findings reveal that probiotics immediately activate anti-inflammatory mechanisms by producing anti-inflammatory cytokines such as interleukin (IL)-4, IL-10, IL-11, and IL-13, and hindering pro-inflammatory cytokines such as IL-1, IL-6, and TNF-α by involving regulatory T cells (Tregs) and T helper cells (Th cells). Several strains of Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus reuteri, Bifidobacterium longum, and Bifidobacterium breve have been listed among the probiotics that are excellent in alleviating various simple to complex ailments. Therefore, the importance of probiotics necessitates robust research to unveil the implications of probiotics, including the potency of strains, the optimal dosages, the combination of probiotics, their habitat in the host, the host response, and other pertinent factors.

Recent advancements in the utilization of ultrasonic technology for the curing of processed meat products: A comprehensive review.

Curation meat products involves multiple stages, including pre-curing processing (thawing, cleaning, and cutting), curing itself, and post-curing processing (freezing, and packaging). Ultrasound are nonthermal processing technology widely used in food industry. This technology is preferred because it reduces the damages caused by traditional processing techniques on food, while simultaneously improving the nutritional properties and processing characteristics of food. The utilization of ultrasonic-assisted curing technology has attracted significant attention within the realm of meat product curing, encouraging extensive research efforts. In terms of curing meat products, ultrasonic-assisted curing technology has been widely studied due to its advantages of accelerating the curing speed, reducing nutrient loss, and improving the tenderness of cured meats. Therefore, this article aims to comprehensively review the application and mechanism of ultrasound technology in various stages of meat product curing. Furthermore, it also elaborates the effects of ultrasonic-assisted curing on the tenderness, water retention, and flavor substances of the meat products during the curing process. Besides, the implication of the ultrasound in the processing of meat curation plays a potent role together with other technologies or methods. The use of ultrasound technology in the process of meat curation was analyzed, which might be a theoretical insight for the industrialization prospects of the meat product.

The Application of Cone Beam Computed Tomography (CBCT) on the Diagnosis and Management of Maxillofacial Trauma.

The assessment and management of facial trauma in an acute setting is one of the core services provided by oral and maxillofacial units in the United Kingdom. Imaging is a pre-requisite for appropriate diagnosis and treatment planning, with a combination of plain radiographs and medical-grade CT being the mainstay. However, the emergence of cone beam CT in recent years has led to its wider applications, including facial trauma assessment. It can offer multi-planar reformats and three-dimensional reconstruction at a much lower radiation dose and financial cost than conventional CT. The purpose of this review is to appraise its potential indications in all anatomical areas of maxillofacial trauma and provide our experience at a level 1 trauma centre.

Exploring the potential of pullulan-based films and coatings for effective food preservation: A comprehensive analysis of properties, activation strategies and applications.

Pullulan is naturally occurring polysaccharide exhibited potential applications for food preservation has gained increasing attention over the last half-century. Recent studies focused on efficient preservation and targeted inhibition using active composite ingredients and advanced technologies. This has led to the emergence of pullulan-based biofilm preservation. This review extensively studied the characteristics of pullulan-based films and coatings, including their mechanical strength, water vapor permeability, thermal stability, and potential as a microbial agent. Furthermore, the distinct characteristics of pullulan, production methods, and activation strategies, such as pullulan derivatization, various compounded ingredients (plant extracts, microorganisms, and animal additives), and other technologies (e.g., ultrasound), are thoroughly studied for the functional property enhancement of pullulan-based films and coatings, ensuring optimal preservation conditions for diverse food products. Additionally, we explore hypotheses that further illuminate pullulan's potential as an eco-friendly bioactive material for food packaging applications. In addition, this review evaluates various methods to improve the efficiency of the film-forming mechanism, such as improving the direct coating process, bioactive packaging films, and implementing layer-by-layer coatings. Finally, current analyses put forward suggestions for future advancement in pullulan-based bioactive films, with the aim of expanding their range of potential applications.

The effects of different hydrocolloids on lotus root starch gelatinization and gels properties.

In this study, xanthan gum (XG), sodium alginate (SA), guar gum (GG), and gum Arabic (GA), were used to modify Lotus root starch (LRS). The incorporation XG, SA, and GG significantly (p < 0.05) influence the swelling power (SP) of LRS, among which the 1.5 % of XG exhibited the highest value of 25.84 g/g at 90 °C. Gelatinization analysis revealed that XG raised the final viscosity (FV) and lowered the breakdown (BD), while SA significantly increased peak viscosity (PV) and BD. Furthermore, GG and GA exhibited a substantial reduction in setback (SB). The incorporation of XG, SA, and GG enhanced the rheological and structural properties (e.g., gel strength and elasticity) of LRS. Particularly, XG demonstrated a more prominent effect, while GA exhibited an opposite trend. Moreover, the structural analyses revealed that hydrophilic colloids have no impact on the functional group and crystal structure of the LRS. However, complex system exhibited the more stable hydrogen bonding. The addition of 1.5 % XG exhibited the most stable hydrogen bonding and highest water binding affinity. Overall, the results demonstrated the effect of different hydrophilic colloids on LRS, offering a theoretical basis for LRS applications and novel insights for the use of starches and hydrocolloids.

Multi-scale ultrasound induced composite coacervates of whey protein and pullulan polysaccharide on emulsion forming and stabilizing mechanisms.

A non-destructive technique known as multi-scale ultrasound (MSU) was employed to modify the emulsion consisting of glycosylated bovine whey protein (WP) and pullulan (Pu). To assess the effect on the structural and emulsifying properties of the WP-Pu, the formulated emulsion, was treated with divergent MSU at (single: 20 kHz, 40 kHz, and 60 kHz; dual: 20-40 kHz, 40-60 kHz, and 20-60 kHz; and tri: 20-40-60 kHz) frequency for a duration of 30 min. The tri-frequency, treated emulsion showed improved emulsifying stability compared to the control and MSU-treated single, and dual-frequency samples, as indicated by the particle size, structural morphology, and adsorbed protein. The molecular docking and numerous spectral analysis provided evidence that WP can undergo successful phenolation. This modified form of WP then interacts with Pu through various forces, including H-bonding and other mechanisms, resulting in the formation of a composite emulsion. The rheological properties revealed that both the control emulsion and the MSU-treated emulsion exhibited non-Newtonian pseudoplastic flow behavior. This behavior is characterized by shear thinning, where the viscosity decreases with increasing shear rate. The shear rates tested ranged from 1 to 300 1/s, additionally, the degree of crystallinity increased from 18.2° to 19.4°. Overall, the tri-frequency effect was most pronounced compared to single and dual-frequency. Ultrasonication, an emerging non-thermal technology, proves to be an efficient approach for the formulation of WP-Pu composites. These composites have significant potential for use in drug delivery systems and functional foods.

Production of Kudzu Starch Gels with Superior Mechanical and Rheological Properties through Submerged Ethanol Exposure and Implications for In Vitro Digestion.

Producing starch gels with superior mechanical attributes remains a challenging pursuit. This research sought to develop a simple method using ethanol exposure to produce robust starch gels. The gels' mechanical properties, rheology, structural characteristics, and digestion were assessed through textural, rheological, structural, and in vitro digestion analyses. Our investigation revealed an improvement in the gel's strength from 62.22 to178.82 g. The thermal transitions were accelerated when ethanol was elevated. The exposure to ethanol resulted in a reduction in syneresis from 11% to 9.5% over a period of 6 h, with noticeable changes in size and color. Rheologically, the dominating storage modulus and tan delta (<0.55) emphasized the gel's improved elasticity. X-ray analysis showed stable B- and V-type patterns after ethanol exposure, with relative crystallinity increasing to 7.9%. Digestibility revealed an ethanol-induced resistance, with resistant starch increasing from 1.87 to 8.73%. In general, the exposure to ethanol played a crucial role in enhancing the mechanical characteristics of kudzu starch gels while simultaneously preserving higher levels of resistant starch fractions. These findings have wide-ranging implications in the fields of confectioneries, desserts, beverages, and pharmaceuticals, underscoring the extensive academic and industrial importance of this study.

A comprehensive review of ultrasonic assisted extraction (UAE) for bioactive components: Principles, advantages, equipment, and combined technologies.

The increasing focus on health and well-being has sparked a rising interest in bioactive components in the food, pharmaceutical, and nutraceutical industries. These components are gaining popularity due to their potential benefits for overall health. The growing interest has resulted in a continuous rise in demand for bioactive components, leading to the exploration of both edible and non-edible sources to obtain these valuable substances. Traditional extraction methods like solvent extraction, distillation, and pressing have certain drawbacks, including lower extraction efficiency, reduced yield, and the use of significant amounts of solvents or resources. Furthermore, certain extraction methods necessitate high temperatures, which can adversely affect certain bioactive components. Consequently, researchers are exploring non-thermal technologies to develop environmentally friendly and efficient extraction methods. Ultrasonic-assisted extraction (UAE) is recognized as an environmentally friendly and highly efficient extraction technology. The UAE has the potential to minimize or eliminate the need for organic solvents, thereby reducing its impact on the environment. Additionally, UAE has been found to significantly enhance the production of target bioactive components, making it an attractive method in the industry. The emergence of ultrasonic assisted extraction equipment (UAEE) has presented novel opportunities for research in chemistry, biology, pharmaceuticals, food, and other related fields. However, there is still a need for further investigation into the main components and working modes of UAEE, as current understanding in this area remains limited. Therefore, additional research and exploration are necessary to enhance our knowledge and optimize the application of UAEE. The core aim of this review is to gain a comprehensive understanding of the principles, benefits and impact on bioactive components of UAE, explore the different types of equipment used in this technique, examine the various working modes and control parameters employed in UAE, and provide a detailed overview of the blending of UAE with other emerging extraction technologies. In conclusion, the future development of UAEE is envisioned to focus on achieving increased efficiency, reduced costs, enhanced safety, and improved reliability. These key areas of advancement aim to optimize the performance and practicality of UAEE, making it a more efficient, cost-effective, and reliable extraction technology.

Ultrasonic and homogenization: An overview of the preparation of an edible protein-polysaccharide complex emulsion.

Emulsion systems are extensively utilized in the food industry, including dairy products, such as ice cream and salad dressing, as well as meat products, beverages, sauces, and mayonnaise. Meanwhile, diverse advanced technologies have been developed for emulsion preparation. Compared with other techniques, high-intensity ultrasound (HIUS) and high-pressure homogenization (HPH) are two emerging emulsification methods that are cost-effective, green, and environmentally friendly and have gained significant attention. HIUS-induced acoustic cavitation helps in efficiently disrupting the oil droplets, which effectively produces a stable emulsion. HPH-induced shear stress, turbulence, and cavitation lead to droplet disruption, altering protein structure and functional aspects of food. The key distinctions among emulsification devices are covered in this review, as are the mechanisms of the HIUS and HPH emulsification processes. Furthermore, the preparation of emulsions including natural polymers (e.g., proteins-polysaccharides, and their complexes), has also been discussed in this review. Moreover, the review put forward to the future HIUS and HPH emulsification trends and challenges. HIUS and HPH can prepare much emulsifier-stable food emulsions, (e.g., proteins, polysaccharides, and protein-polysaccharide complexes). Appropriate HIUS and HPH treatment can improve emulsions' rheological and emulsifying properties and reduce the emulsions droplets' size. HIUS and HPH are suitable methods for developing protein-polysaccharide forming stable emulsions. Despite the numerous studies conducted on ultrasonic and homogenization-induced emulsifying properties available in recent literature, this review specifically focuses on summarizing the significant progress made in utilizing biopolymer-based protein-polysaccharide complex particles, which can provide valuable insights for designing new, sustainable, clean-label, and improved eco-friendly colloidal systems for food emulsion. PRACTICAL APPLICATION: Utilizing complex particle-stabilized emulsions is a promising approach towards developing safer, healthier, and more sustainable food products that meet legal requirements and industrial standards. Moreover, the is an increasing need of concentrated emulsions stabilized by biopolymer complex particles, which have been increasingly recognized for their potential health benefits in protecting against lifestyle-related diseases by the scientific community, industries, and consumers.

Characterization of Nano-SiO2/Zein Film Prepared Using Ultrasonic Treatment and the Ability of the Prepared Film to Resist Different Storage Environments.

This study has developed, ultrasound-assisted, a novel food packaging film (U-zein/SiO2) for food packaging applications. Incorporating an optimal concentration of 18 mg/mL of nano-SiO2 and subjecting the film to 10 min of ultrasonic treatment resulted in a remarkable increase of 32.89% in elongation at break and 55.86% in tensile strength. In addition, the incorporation of nano-SiO2 effectively reduces the water content and solubility of the composite film, resulting in improved water/oxygen barrier properties. These physiochemical properties were further improved with the application of ultrasound. The analysis of attenuated total reflectance-Fourier transform infrared, X-ray diffraction, differential scanning calorimetry, and scanning electronic microscope demonstrated that the ultrasound treatment improved the hydrogen bonds, improved thermal stability, molecular arrangement, structure stability, and intermolecular compatibility of the composite film, resulting in enhanced physio-mechanical properties of the film. In addition, the ultrasound treatment led to a smoother film surface and reduced the pores on the film's cross-section. Moreover, the U-zein/SiO2 film exhibited excellent mechanical and water/oxygen barrier properties in different storage environments over a period of 30 days. These results offer sound theoretical support for the practical application of the prepared preservative film.

The ASNR-MICCAI Brain Tumor Segmentation (BraTS) Challenge 2023: Intracranial Meningioma.

Meningiomas are the most common primary intracranial tumor in adults and can be associated with significant morbidity and mortality. Radiologists, neurosurgeons, neuro-oncologists, and radiation oncologists rely on multiparametric MRI (mpMRI) for diagnosis, treatment planning, and longitudinal treatment monitoring; yet automated, objective, and quantitative tools for non-invasive assessment of meningiomas on mpMRI are lacking. The BraTS meningioma 2023 challenge will provide a community standard and benchmark for state-of-the-art automated intracranial meningioma segmentation models based on the largest expert annotated multilabel meningioma mpMRI dataset to date. Challenge competitors will develop automated segmentation models to predict three distinct meningioma sub-regions on MRI including enhancing tumor, non-enhancing tumor core, and surrounding nonenhancing T2/FLAIR hyperintensity. Models will be evaluated on separate validation and held-out test datasets using standardized metrics utilized across the BraTS 2023 series of challenges including the Dice similarity coefficient and Hausdorff distance. The models developed during the course of this challenge will aid in incorporation of automated meningioma MRI segmentation into clinical practice, which will ultimately improve care of patients with meningioma.

Preparation and characterization of ultrasound-assisted essential oil-loaded nanoemulsions stimulated pullulan-based bioactive film for strawberry fruit preservation.

Ultrasound is a high-energy approach that can help with homogenization and dispersion in cavitation. In this study, nanoemulsions of curcumin and orange essential oil were prepared with ultrasound treatment at different times. The ultrasound-treated nanoemulsions for 10 min exhibited the smallest droplet size, the best storage, and higher thermal stability. The pullulan-based film with ultrasound-assisted nanoemulsions exhibited improved water vapor permeability and moisture content and the highest tensile strength and elongation at break. The structural analysis showed that ultrasonic treatment enhanced the H-bond, resulting in a more orderly molecular arrangement and intermolecular compatibility. Furthermore, the bioactive film had the maximum oil retention time. It possessed excellent bacteriostatic properties against Escherichia coli and Staphylococcus aureus due to the smallest oil droplets and uniform distribution in the film matrix. Besides, the weight loss and deterioration of the strawberry fruit were effectively reduced, thus prolonging the shelf life.

Effects of multi-frequency ultrasound on sodium caseinate/pectin complex: Emulsifying properties, interaction force, structure and correlation.

This study aimed to improve the emulsification properties of the sodium caseinate (Cas) and pectin (Pec) complex using multi-frequency power ultrasound to regulate the complexation of Cas and Pec. The results revealed that optimal ultrasonic treatment (Frequency 60 kHz, power density 50 W/L, and time 25 min) led to a 33.12 % increase in emulsifying activity (EAI) and a 7.27 % increase in emulsifying stability index (ESI) of the Cas-Pec complex. Our results demonstrated that electrostatic interactions and hydrogen bonds were the main driving forces for complex formation, and these were reinforced by ultrasound treatment. Moreover, it was observed that ultrasonic treatment improved the surface hydrophobicity, thermal stability, and secondary structure of the complex. Scanning electron microscopy and atomic force microscopy analyses revealed that the ultrasonically prepared Cas-Pec complex had a dense, uniform spherical structure with reduced surface roughness. It was further confirmed that the complex's emulsification properties were highly correlated with its physicochemical and structural properties. Multi-frequency ultrasound changes the interaction by regulating protein structure and ultimately acting on the interfacial adsorption behavior of the complex. This work contributes to expanding the role of multi-frequency ultrasound in modifying the emulsification properties of the complex.

Preparation technology and preservation mechanism of γ-CD-MOFs biaological packaging film loaded with curcumin.

To prevent food spoilage caused by microbial infection, the development of an environmentally friendly antimicrobial preservation material is crucial. Here, the microporous γ-CD-MOFs was utilized to encapsulate the hydrophobic active substance curcumin, resulting in the preparation of a non-toxic antimicrobial material (Cur-CD-MOFs). The results revealed that curcumin encapsulation in Cur-CD-MOFs occurred primarily in the carbonyl group, benzene ring, and enolic side ring of curcumin. The Cur-CD-MOFs had a 100% bactericidal effect on Escherichia coli and Staphylococcus aureus at 4 h and 8 h, and a strong inhibitory effect on aerial mycelium of Penicillium expansum and Botrytis cinerea. Furthermore, the incorporation of Cur-CD-MOFs improved the Pul/Tre film barrier and mechanical properties. The effectiveness of Cur-CD-MOFs-Pul/Tre in retaining fruit freshness was validated using Centennial Seedless grapes. This study confirmed that Cur-CD-MOFs is a promising antibacterial material, and Cur-CD-MOFs-Pul/Tre will be a potent candidate for food preservation.

Assessing impact of agroecological interventions in Niger through remotely sensed changes in vegetation.

Water scarcity is a major challenge in the Sahel region of West Africa. Water scarcity in combination with prevalent soil degradation has severely reduced the land productivity in the region. The decrease in resiliency of food security systems of marginalized population has huge societal implications which often leads to mass migrations and conflicts. The U.S. Agency for International Development (USAID) and development organizations have made major investments in the Sahel to improve resilience through land rehabilitation activities in recent years. To help restore degraded lands at the farm level, the World Food Programme (WFP) with assistance from USAID's Bureau for Humanitarian Assistance supported the construction of water and soil retention structures called half-moons. The vegetation growing in the half-moons is vitally important to increase agricultural productivity and feed animals, a critical element of sustainable food security in the region. This paper investigates the effectiveness of interventions at 18 WFP sites in southern Niger using vegetative greenness observations from the Landsat 7 satellite. The pre - and post-intervention analysis shows that vegetation greenness after the half-moon intervention was nearly 50% higher than in the pre-intervention years. The vegetation in the intervened area was more than 25% greener than the nearby control area. Together, the results indicate that the half-moons are effective adaptations to the traditional land management systems to increase agricultural production in arid ecosystems, which is evident through improved vegetation conditions in southern Niger. The analysis shows that the improvement brought by the interventions continue to provide the benefits. Continued application of these adaptation techniques on a larger scale will increase agricultural production and build resilience to drought for subsistence farmers in West Africa. Quantifiable increase in efficacy of local-scale land and water management techniques, and the resulting jump in large-scale investments to scale similar efforts will help farmers enhance their resiliency in a sustainable manner will lead to a reduction in food security shortages.

Non-thermal effects of microwave irradiation alleviates postharvest chilling injury of peach fruit by retarding phenolic accumulation and enhancing membrane stability.

Postharvest chilling injury (CI) of fruit, including peaches, is a huge challenge to horticultural product preservation. Microwave irradiation can be used as a physiological regulator due to the thermal effects; however, its non-thermal effects on the CI of postharvest fruit remain unclear. Thus, the physiological attributes and metabolisms involving phenolics, fatty acids, and sugars were compared between 'Zhongtao No.9' peaches treated with microwave irradiation at 45.5 W for different durations and control. Microwave treatment especially at 45.5 W for 7 min without inducing thermal effects could significantly inhibit internal browning caused by CI, concomitant with reduced total phenolic content. Moreover, the maintenance of membrane stability was indicated by a boosted double bond index, which may be attributed to the inhibition of membrane lipid degradation, and sucrose accumulation. In summary, the non-thermal effects of microwave irradiation contribute to CI alleviation through restraining phenolic content and maintaining membrane stability in peach fruit.

Insights into ultrasonic treatment on the properties of pullulan/oat protein/nisin composite film:mechanical, structural and physicochemical properties.

This study developed two novel food packaging films, oat protein/pullulan (Op/Pul) and Nisin-loaded oat protein/pullulan (Nis@Op/Pul) films. Ultrasound was introduced to improve its mechanical, structural and physicochemical properties. The Op/Pul film has lower light transmittance, water vapour and oxygen permeability (OP) and improved film uniformity than pure oat protein and pullulan film. The addition of Nisin led to a significant decrease in the composite films' transparency, moisture content, and total soluble matter (TSM). The ultrasound treatment significantly increased the elongation at break and transparency of Nis@Op/Pul film by 18.37% and 8.03% and decreased its TSM and OP by 8.33% and 2.78%, respectively, compared to the conventional method. The structure analysis shows ultrasound enhances intermolecular hydrogen bonding, reduces the crystallinity and formed a more regular, uniform surface. Moreover, the Nis@Op/Pul film prepared by ultrasound treatment could effectively delay the decay and deterioration of fresh strawberries and prolong their shelf life.

Ultrasound-assisted development and characterization of novel polyphenol-loaded pullulan/trehalose composite films for fruit preservation.

A novel food packaging film was developed by incorporating a tea polyphenols-loaded pullulan/trehalose (TP@Pul/Tre) into a composite film with ultrasound-assisted treatment of dual-frequency (20/35 kHz, 40 W/L) for 15 min to assess the physicochemical and mechanical properties of a composite film. The optimized ultrasound-assisted significantly increases elongation at break, tensile strength, and improves the composite film's UV/water/oxygen barrier properties. Structure analysis using attenuated total reflectance-Fourier transform infrared, X-ray diffraction and thermal stability revealed that these improvements were achieved through ultrasound-enhanced H-bonds, more ordered molecular arrangements, and good intermolecular compatibility. Besides, the ultrasound-assisted TP@Pul/Tre film has proven to have good antibacterial performance against Escherichia coli and Staphylococcus aureus, with approximately 100 % lethality at 4 h and 8 h, respectively. Moreover, the ultrasound-assisted TP@Pul/Tre film effectively delayed moisture loss, oxidative browning, decay, and deterioration in fresh-cut apples and pears, thereby extending their shelf life. Thus, ultrasound has proved to be an effective tool for improving the quality of food packaging films, with a wide range of applications.

Functional analysis of the dihydroflavonol 4-reductase family of Camellia sinensis: exploiting key amino acids to reconstruct reduction activity.

Anthocyanins and proanthocyanidins (PAs) are important types of flavonoids, plant secondary metabolites with a wide range of industrial and pharmaceutical applications. DFR (dihydroflavonol 4-reductase) is a pivotal enzyme that plays an important role in the flavonoid pathway. Here, four CsDFR genes were isolated from Camellia sinensis, and their overexpression was analyzed in vitro and in vivo. Based on transcription and metabolic analyses, CsDFR expression was closely consistent with catechins and PAs accumulation. Moreover, enzyme activity analyses revealed that the two recombinant proteins CsDFRa and CsDFRc exhibited DFR activity, converting dihydroflavonols into leucoanthocyanins in vitro, but CsDFRb1 and CsDFRb3 did not. CsDFRa and CsDFRc overexpression in AtDFR mutants (tt3) revealed that CsDFRs are involved in the biosynthesis of anthocyanins and PAs, as CsDFRa and CsDFRc restored not only the purple petiole phenotype but also the seed coat color. Site-directed mutagenesis revealed that the two amino acid residues S117 and T123 of CsDFRa play a prominent role in controlling DFR reductase activity. Enzymatic assays indicated that CsDFRa and CsDFRc exhibited a higher affinity for DHQ and DHK, respectively, whereas CsDFRb1N120S and CsDFRb1C126T exhibited a higher affinity for DHM. Our findings comprehensively characterize the DFRs from C. sinensis and shed light on their critical role in metabolic engineering.