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1.
Int J Biol Macromol ; : 136832, 2024 Oct 24.
Artículo en Inglés | MEDLINE | ID: mdl-39461627

RESUMEN

Equol is an isoflavone-derived metabolite known to exhibit strong estrogenic and antioxidant activities. The aim of this paper is twofold: first, to confirm the anticancer potential of equol against colorectal cancer, and second, to reveal the underlying mechanisms. After treatment with 40 µg/mL equol, cell proliferation, cell migration, and colony formation of HCT116 colon cancer cells were inhibited. Network pharmacology and transcriptomics analysis revealed the downregulation of genes related to DNA replication (CCND1, E2F1, CDC6, CDC45, MCM4), leading to G1/S cell cycle arrest and the induction of cell apoptosis, which was confirmed by flow cytometry. Genes associated with the G2-to-M transition (CDK1, CCNA2, CCNB1) were also downregulated. In addition, equol downregulated genes (FOXM1 and ASPM) that control cell migration and invasion. Our data indicate that equol can inhibit colorectal cancer by targeting multiple pathways, suggesting its potential as a key component in the adjuvant treatment of colorectal cancer.

2.
Food Chem X ; 24: 101725, 2024 Dec 30.
Artículo en Inglés | MEDLINE | ID: mdl-39431207

RESUMEN

This study employed extraction methods, namely acid, alkaline, ultrasonic-assisted, hot-water, and dual enzyme-assisted extraction to extract polysaccharides from Tricholoma mongolicum Imai (TMIPs), and investigated them for intestinal digestion and fecal fermentation in vitro. Furthermore, using fructo-oligosaccharide as a positive prebiotic control, the impact of these TMIPs as carbon sources on the growth of Lactobacillus and Bifidobacterium in liquid culture was assessed. The results showed that all fractions transit through the gastrointestinal tract without degradation. Additionally, compared to the control group, the five polysaccharides significantly promoted the growth of probiotics, with a significant increase in short-chain fatty acid production after 48 h of fermentation. Furthermore, all five polysaccharides modulated the composition of gut microbiota. This offers theoretical guidance in the rational advancement of functional products derived from edible mushrooms, aiming to enhance gastrointestinal health in humans.

3.
Front Pharmacol ; 15: 1367417, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39224778

RESUMEN

Background: Hepatocellular carcinoma currently has the third highest mortality rate in the world. Patients with hepatocellular carcinoma are on the rise and at a younger age, but research into the pharmacological effects of cancer is mostly single-component, and natural plant products can have additive or synergistic effects that can better amplify the effects of intervention in cancer. Aim: To evaluate the synergistic therapeutic effects of 6-shogaol and curcumin against hepatocellular carcinoma line HepG2 cells. Methods: In this study, a network pharmacology approach was used to predict and validate the mol ecular targets and pathways of the hepatocellular carcinoma (HCC) of 6-shogaol and curcumin in combination and to investigate their mechanism of action. The results were also validated by cellular assays. HepG2 cells were treated with 6-shogaol and curcumin as well as the combination of the two. The combination index of 6-shogaol and curcumin in HepG2 cells was calculated using Compusyn software according to the Chou-Talalay equation. The synergistic anti-cancer effect was next investigated by MTT assay, apoptosis assay and cell cycle assay. The combined anti-hepatocellular carcinoma effect of the Ras-mediated PI3K/AKT and MAPK signalling pathways was analysed using protein blotting assays. Results: A network pharmacology-based screening identified 72 core targets of 6-curcumin and curcumin in hepatocellular carcinoma, and predicted that the main signalling pathway is the Ras signalling pathway. The anti-cancer effects of 6-shogaol and curcumin were validated in cell-based assays and the optimal synergistic concentrations of 5 µmoL/L for 6-shogaol and 30 µmoL/L for curcumin were determined. 6-shogaol and curcumin synergistically blocked the cell cycle in the G2/M phase and promoted apoptosis. Immunoblot analysis confirmed for the first time the combined action of both in down-regulating the Ras-mediated PI3K/AKT and MAPK signaling pathways. In addition, 6-shogaol and curcumin acting together downregulated Cyclin-B, CDK-1, Bcl-2, and upregulated BAX. Conclusion: 6-shogaol and curcumin act synergistically to alter the morphology of hepatocellular carcinoma cells, block the cell cycle in the G2/M phase, inhibit proliferation and division, and effectively promote late apoptosis. The combined action of these two components provides a theoretical basis for the further development of novel anti-liver cancer products.

4.
Food Chem ; 463(Pt 1): 141442, 2024 Sep 25.
Artículo en Inglés | MEDLINE | ID: mdl-39342685

RESUMEN

This study incorporated purple sweet potato anthocyanin (PSPA) and silver-nanoparticles (AgNPs) into the chitosan/polyvinyl alcohol film matrix (PVA/CS) to successfully prepare a composite film, which effectively inhibited bacterial growth and indicated product freshness. The addition of AgNPs and PSPA led to a dense structure of the film, which effectively enhanced its physical properties, barrier properties and functional properties. The incorporation of PSPA made the composite film highly pH-sensitive, which exhibited distinct color changes in varying pH solutions. The PVA/CS-AgNPs-PSPA10 composite film with PSPA and AgNPs resulted the shelf life of strawberries to 13 days at 4 °C, which effectively reduced strawberry breathing during storage. Additionally, such composite film changed color from purple to yellow-purple, indicating the deterioration of strawberries. It also showed an antibacterial indication through its excellent antibacterial property and freshness indication performance, which demonstrated its significance in developing antibacterial indicator composite packaging materials for fruits and vegetables preservation.

5.
Int J Biol Macromol ; 278(Pt 2): 134758, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39151846

RESUMEN

Steam explosion (SE) is a potential method to modify pectin structure, which might be connected to its emulsifying characteristics and the bioavailability of encapsulated polymethoxyflavone like nobiletin. However, the relationship between SE-modified pectin and the bioavailability of encapsulated nobiletin is still unclear. In this study, nobiletin-loaded emulsion was fabricated using citrus pectin modified with SE (0.15-0.9 MPa, 3 min) as emulsifier for in vitro digestion study, and the transport and absorption of nobiletin in Caco-2 cells to investigate the bioavailability-promoting effect. The results showed that SE treatment lowered the droplet size of emulsion from 21.38 ± 2.30 µm to 2.14 ± 0.12 µm, enhanced the nobiletin encapsulation efficiency from 23.73 ± 0.78% to 86.27 ± 3.81%, improved the nobiletin bioaccessibility in vitro from 2.48 ± 0.10% to 25.42 ± 0.10% and increased the intracellular accumulation of nobiletin by over 10 times, even higher than that of Tween 80. In conclusion, pectin from SE-treated citrus peel exhibited good emulsion properties and bioavailability-promoting effect in vitro of nobiletin.


Asunto(s)
Disponibilidad Biológica , Citrus , Emulsiones , Flavonas , Pectinas , Vapor , Flavonas/química , Flavonas/farmacología , Flavonas/farmacocinética , Pectinas/química , Pectinas/farmacología , Humanos , Emulsiones/química , Células CACO-2 , Citrus/química
6.
Int J Biol Macromol ; 277(Pt 1): 134094, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39059525

RESUMEN

Leuconostoc citreum JZ-002 was extracted from artisanal orange wine. This strain was used to synthesize dextran with a purification extraction of 27.9 g/L. The resulting dextran had a molecular weight of 2.45 × 106 Da. A significant portion, amounting to 64 % of the structure, is constituted by the main chain, with α-(1,6) glycosidic bonds acting as the linkages. In contrast, the branched chain, comprising 34 % of the entire molecule, is characterized by the presence of α-(1,3) glycosidic bonds. The dextransucrase DsrB, believed to be accountable for the formation of the dextran backbone, was successfully cloned into the pET-28a-AcmA vector. The recombinant expression of the enzyme was achieved. Purified recombinant enzymes and immobilized in a single go using the gram-positive enhancer matrix (GEM). The maximum yield of dextran produced by suchimmobilized enzyme was 191.9 g/L. The composition featured a dextran connected via α-(1,6) glycosidic linkages. Molecular weight controlled synthesis was achieved with sucrose concentrations of 100-2000 mM and enzyme concentrations of 320-1280 U. The Mw of the synthesized dextran extended from 4680 to 1,320,000 Da. By controlling the ratio between enzyme concentration and sucrose concentration, dextrans with diverse Mw can be enzymatically generated.


Asunto(s)
Dextranos , Glucosiltransferasas , Leuconostoc , Dextranos/química , Dextranos/biosíntesis , Enzimas Inmovilizadas/química , Enzimas Inmovilizadas/metabolismo , Glucosiltransferasas/química , Glucosiltransferasas/genética , Glucosiltransferasas/metabolismo , Leuconostoc/enzimología , Peso Molecular , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Sacarosa/química
7.
Compr Rev Food Sci Food Saf ; 23(4): e13364, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38847746

RESUMEN

Kefir milk, known for its high nutritional value and health benefits, is traditionally produced by fermenting milk with kefir grains. These grains are a complex symbiotic community of lactic acid bacteria, acetic acid bacteria, yeasts, and other microorganisms. However, the intricate coexistence mechanisms within these microbial colonies remain a mystery, posing challenges in predicting their biological and functional traits. This uncertainty often leads to variability in kefir milk's quality and safety. This review delves into the unique structural characteristics of kefir grains, particularly their distinctive hollow structure. We propose hypotheses on their formation, which appears to be influenced by the aggregation behaviors of the community members and their alliances. In kefir milk, a systematic colonization process is driven by metabolite release, orchestrating the spatiotemporal rearrangement of ecological niches. We place special emphasis on the dynamic spatiotemporal changes within the kefir microbial community. Spatially, we observe variations in species morphology and distribution across different locations within the grain structure. Temporally, the review highlights the succession patterns of the microbial community, shedding light on their evolving interactions.Furthermore, we explore the ecological mechanisms underpinning the formation of a stable community composition. The interplay of cooperative and competitive species within these microorganisms ensures a dynamic balance, contributing to the community's richness and stability. In kefir community, competitive species foster diversity and stability, whereas cooperative species bolster mutualistic symbiosis. By deepening our understanding of the behaviors of these complex microbial communities, we can pave the way for future advancements in the development and diversification of starter cultures for food fermentation processes.


Asunto(s)
Kéfir , Simbiosis , Kéfir/microbiología , Simbiosis/fisiología , Microbiota/fisiología , Fermentación , Microbiología de Alimentos
8.
Food Chem ; 455: 139884, 2024 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-38865845

RESUMEN

Glycation is a promising approach to enhance protein gel characteristics in the food industry. The impact of oyster myofibrillar protein (MP) being glycosylated with six oligosaccharides (dextran [Dex]-1 kDa, 5 kDa, 6 kDa, and 10 kDa, xylan [Xyla], and xyloglucan [Xyg]) on structural properties, aggregation behavior and gel properties was investigated in this study. The findings demonstrated that oligosaccharides significantly increased the glycation degree of MP by forming a stable tertiary conformation, increasing the contents of the disulfide bond and hydrogen bonds. Additionally, particle sizes decreased and solubility increased after glycation, improving the gel's strength, water-holding capacity, thermal stability, elastic modulus, and ordered network layout. It was determined that MP-Dex 5 had the best gel properties. The gel strength and water holding capacity of MP-Dex 5 increased by 70.59% and 32.27%, respectively. Molecular dynamics simulations results showed van der Waals energy and electrostatic interactions favor myosin binding to Dex or Xyla units. This study will provide insights into the relationship between molecular structure, aggregation behavior and gel property of oyster MP-oligosaccharide couples, and expand the application of oyster MP in food gels.


Asunto(s)
Crassostrea , Geles , Oligosacáridos , Animales , Oligosacáridos/química , Geles/química , Crassostrea/química , Proteínas Musculares/química , Simulación de Dinámica Molecular , Glicosilación , Solubilidad
9.
Food Chem ; 457: 140151, 2024 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-38901353

RESUMEN

The characteristic aroma compounds of Chinese steamed bread (CSB) fermented with different starters were studied using HS-SPME-GC/MS, aroma recombination and omission experiments. The dynamic changes of the microbiota and their function and metabolites during fermentation were analyzed using metagenomics and non-targeted metabolomics. Forty-nine volatile flavor compounds were identified, while 5 characteristic aroma-active compounds were investigated in CSB fermented with commercial dry yeast (AQ-CSB), and 10 were investigated in CSB fermented with traditional starter (NY-CSB). Microbial structure and function analysis showed that Saccharomyces cerevisiae dominated during AQ-CSB fermentation and contributed >95% to its KEGG pathways, while Pediococcus pentosaceus, unclassified Pediococcus, Lactobacillus plantarum, Lactobacillus brevis and unclassified Lactobacillus were predominant in NY-CSB and together had an ~96% contribution to these pathways. NY-CSB showed higher metabolic activity during fermentation, and the characteristic metabolites were mainly involved in carbohydrate, amino acid and lipid metabolism. The characteristic aroma compounds were identified and increased the understanding of the contributions of the microbiota. This may be useful for designing starter cultures that produce CSB with desirable aroma properties.


Asunto(s)
Pan , Fermentación , Odorantes , Saccharomyces cerevisiae , Compuestos Orgánicos Volátiles , Bacterias/metabolismo , Bacterias/genética , Bacterias/clasificación , Bacterias/aislamiento & purificación , Pan/análisis , Pan/microbiología , Aromatizantes/metabolismo , Aromatizantes/química , Cromatografía de Gases y Espectrometría de Masas , Microbiota , Odorantes/análisis , Saccharomyces cerevisiae/metabolismo , Compuestos Orgánicos Volátiles/metabolismo , Compuestos Orgánicos Volátiles/química , Compuestos Orgánicos Volátiles/análisis
10.
Food Chem ; 457: 140197, 2024 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-38941907

RESUMEN

The purpose of this study is to investigate the mechanism by which 6-shogaol ameliorates hepatic steatosis via miRNA-mRNA interaction analysis. C57BL/6 J mice were fed a high-fat diet (HFD) for 12 weeks, during which 6-shogaol was administered orally. The liver lipid level, liver function and oxidative damage in mice were evaluated. mRNA sequencing, miRNA sequencing, and RT-qPCR were employed to compare the expression profiles between the HFD group and the 6-shogaol-treated group. High-throughput sequencing was used to construct the mRNA and miRNA libraries. Target prediction and integration analysis identified eight potential miRNA-mRNA pairs involved in hepatic steatosis, which were subsequently validated in liver tissues and AML12 cells. The findings revealed that 6-shogaol modulates the miR-3066-5p/Grem2 pathway, thereby improving hepatic steatosis. This study provides new insights into the mechanisms through which 6-shogaol alleviates hepatic steatosis, establishing a foundation for future research on natural active compounds for the treatment of metabolic diseases.


Asunto(s)
Catecoles , Dieta Alta en Grasa , Ratones Endogámicos C57BL , MicroARNs , Animales , MicroARNs/genética , MicroARNs/metabolismo , Ratones , Dieta Alta en Grasa/efectos adversos , Masculino , Catecoles/farmacología , Catecoles/administración & dosificación , Humanos , Hígado/metabolismo , Hígado/efectos de los fármacos , Hígado Graso/genética , Hígado Graso/metabolismo , Hígado Graso/tratamiento farmacológico , Hígado Graso/prevención & control , Hígado Graso/etiología
11.
Food Chem ; 452: 139532, 2024 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-38705120

RESUMEN

This study aimed to better understand whether and how the reactive 1,2-dicarbonyl precursors of advanced glycation end products (AGEs), glyoxal (GO) and methylglyoxal (MGO), cross the intestinal barrier by studying their transport in the in vitro Caco-2 transwell system. The results reveal that GO, MGO and Nε-(carboxymethyl)lysine (CML), the latter studied for comparison, are transported across the intestinal cell layer via both active and passive transport and accumulate in the cells, albeit all to a limited extent. Besides, the transport of the dicarbonyl compounds was only partially affected by the presence of amino acids and protein, suggesting that scavenging by a food matrix will not fully prevent their intestinal absorption. Our study provides new insights into the absorption of the two major food-borne dicarbonyl AGE precursors and provides evidence of their potential systemic bioavailability but also of factors limiting their contribution to the overall exposome.


Asunto(s)
Productos Finales de Glicación Avanzada , Glioxal , Piruvaldehído , Humanos , Células CACO-2 , Productos Finales de Glicación Avanzada/metabolismo , Productos Finales de Glicación Avanzada/química , Piruvaldehído/metabolismo , Glioxal/metabolismo , Glioxal/química , Modelos Biológicos , Transporte Biológico , Absorción Intestinal
12.
Food Chem ; 450: 139311, 2024 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-38636377

RESUMEN

Gold nanoparticles (AuNPs)-based immunochromatographic assay has gained popularity as a rapid detection method for food hazards. Synthesizing highly stable AuNPs in a rapid, simple and environmentally friendly manner is a key focus in this field. Here, we present a green microfluidic strategy for the rapid, automated, and size-controllable synthesis of pepsin-doped AuNPs (AuNPs@Pep) by employing glucose-pepsin as a versatile reducing agent and stabilizer. Through combining the colorimetric and photothermal (PoT) properties of AuNPs@Pep, both "signal-off" and "signal-on" formats of microfluidic paper analytical devices (PADs) were developed for detection of a small molecule antibiotic, florfenicol, and an egg allergen, ovalbumin. Compared to the colorimetric mode, a 4-fold and 3-fold improvement in limit of detection was observed in the "signal-off" detection of florfenicol and the "signal-on" detection of ovalbumin, respectively. The results demonstrated the practicality of AuNPs@Pep as a colorimetric/PoT dual-readout probe for immunochromatographic detection of food hazards at different molecular scales.


Asunto(s)
Colorimetría , Contaminación de Alimentos , Oro , Tecnología Química Verde , Nanopartículas del Metal , Colorimetría/métodos , Contaminación de Alimentos/análisis , Oro/química , Límite de Detección , Nanopartículas del Metal/química , Ovalbúmina/química , Pepsina A/química , Tianfenicol/análogos & derivados
13.
Int J Biol Macromol ; 266(Pt 2): 131090, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38537858

RESUMEN

In this study, modified sea Cucumber Peptides (SCP) were prepared by reacting with xylooligosaccharide (XOS) and alginate oligosaccharides (AOS) via glycation. Free radical inhibitory and inhibition of oxidative stress of modified SCP was evaluated using human hepatocellular carcinoma (HepG2) cells and zebrafish embryos. LC-MS analysis revealed that SCPs mainly consist of 40 active peptides, with an average molecular weight of 1122.168 Da and an average length of 11 amino acid residues. For amino acid composition, L-Asparagine, L-Methionine, and L-Aspartic Acid were dominant amino acids in SCP. The result showed that the antioxidant ability of SCP against 2,2-Diphenyl-1-picrylhydrazyl (DPPH), superoxide anion radical (O-2), and Hydroxyl Radical (OH) was significantly improved after modification. In HepG2 cells, the modified SCP showed stronger protection than native SCP native against H2O2-induced oxidative stress by enhancing cell viability and reducing radical oxygen species (ROS) generation. The inhibition effect of SCP was increased after modification with XOS and AOS by 13 % and 19 % respectively. Further studies displayed that the activity of antioxidative enzymes, including Superoxide dismutase (SOD), Glutathione Peroxidase (GPx), and catalase (CAT), was remarkably enhanced, whereas malondialdehyde (MDA) level was reduced compared with native SCP and H2O2-treated groups, thus, improving the intracellular antioxidant defenses. The gene expression analysis showed that the mechanism underlying the modified SCP protective effect may be linked with the capability to regulate Nuclear factor-erythroid factor 2-related factor 2 (NRF2) gene expression. The protective effect of modified SCP against H2O2 in vitro was confirmed in vivo by reduced toxicity in zebrafish embryos via improvement of mortality rate, hatching rate, heart beating rate, and deformities of the zebrafish model. However, SCPAOS conjugate displayed greater antioxidant potentials compared to the SCPXOS, the different effects between SCPAOS and SCPXOS could be due to their different antioxidant activity. Thus, modified SCP could be potentially used as a novel nutraceutical in the preparation of anti-aging food and medicine.


Asunto(s)
Antioxidantes , Peróxido de Hidrógeno , Estrés Oxidativo , Péptidos , Pepinos de Mar , Pez Cebra , Animales , Células Hep G2 , Peróxido de Hidrógeno/farmacología , Humanos , Estrés Oxidativo/efectos de los fármacos , Pepinos de Mar/química , Péptidos/farmacología , Péptidos/química , Antioxidantes/farmacología , Antioxidantes/química , Especies Reactivas de Oxígeno/metabolismo , Supervivencia Celular/efectos de los fármacos , Catalasa/metabolismo
14.
Foods ; 13(6)2024 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-38540878

RESUMEN

A bacteria capable of degrading aflatoxin M1 (AFM1) was isolated from African elephant manure. It was identified as Bacillus pumilus by 16s rDNA sequencing and named B. pumilusE-1-1-1. Compared with physical and chemical methods, biological methods have attracted much attention due to their advantages, such as thorough detoxification, high specificity, and environmental friendliness. This work aimed to study the effects of a recombinant catalase (rCAT) from B. pumilusE-1-1-1 on the degradation of AFM1 in pattern solution. The degradation mechanism was further explored and applied to milk and beer. Kinetic Momentum and Virtual Machine Maximum values for rCAT toward AFM1 were 4.1 µg/mL and 2.5 µg/mL/min, respectively. The rCAT-mediated AFM1 degradation product was identified as C15H14O3. Molecular docking simulations suggested that hydrogen and pi bonds played major roles in the steadiness of AFM1-rCAT. In other work, compared with identical density of AFM1, survival rates of Hep-G2 cells incubated with catalase-produced AFM1 degradation products increased by about 3 times. In addition, degradation rates in lager beer and milk were 31.3% and 47.2%, respectively. Therefore, CAT may be a prospective substitute to decrease AFM1 contamination in pattern solution, milk, and beer, thereby minimizing its influence on human health.

15.
Food Chem ; 447: 138985, 2024 Jul 30.
Artículo en Inglés | MEDLINE | ID: mdl-38507952

RESUMEN

Myofibrillar protein (MP) is susceptible to the effect of ionic strength and ultra-high pressure (UHP) treatment, respectively. However, the impact of UHP combined with ionic strength on the structure and in vitro digestibility of MP from scallop mantle (Patinopecten yessoensis) is not yet clear. Therefore, it is particularly important to analyze the structural properties and enhance the in vitro digestibility of MP by NaCl and UHP treatment. The findings demonstrated that as ionic strength increased, the α-helix and ß-sheet gradually transformed into ß-turn and random coil. The decrease of endogenous fluorescence intensity indicated the formation of a more stable tertiary structure. Additionally, the exposure of internal sulfhydryl groups increased the amount of total sulfhydryl content, and reactive sulfhydryl groups gradually transformed into disulfide bonds. Moreover, it reduces aggregation through increased solubility, decreased turbidity, particle sizes, and a relatively dense and uniform microstructure. When MP from the scallop mantle was treated with 0.5 mol/L ionic strength and 200 MPa UHP treatment, it had the highest solubility (90.75 ± 0.13%) and the lowest turbidity (0.41 ± 0.03). The scallop mantle MP with NaCl of 0.3 mol/L and UHP treatment had optimal in vitro digestibility (95.14 ± 2.01%). The findings may offer a fresh perspectives for developing functional foods for patients with dyspepsia and a theoretical foundation for the comprehensive utilization of scallop mantle by-products with low concentrations of NaCl.


Asunto(s)
Pectinidae , Cloruro de Sodio , Animales , Humanos , Cloruro de Sodio/metabolismo , Proteínas/química , Pectinidae/química , Cloruro de Sodio Dietético
16.
Anal Chim Acta ; 1293: 342283, 2024 Mar 08.
Artículo en Inglés | MEDLINE | ID: mdl-38331551

RESUMEN

Recombinant antibody-based immunoassays have emerged as crucial techniques for detecting antibiotic residues in food samples. Developing a stable recombinant antibody production system and enhancing detection sensitivity are crucial for their biosensing applications. Here, we bioengineered a single-chain fragment variable (scFv) antibody to target chloramphenicol (CAP) using both Bacillus subtilis and HEK 293 systems, with the HEK 293-derived scFv demonstrating superior sensitivity. Computational chemistry analyses indicated that ASP-99 and ASN-102 residues in the scFv play key roles in antibody recognition, and the hydroxyl group near the benzene ring of the target molecule is critical for in antibody binding. Furthermore, we enhanced the scFv's biosensing sensitivity using an HCR-CRISPR/Cas12a amplification strategy in a streptavidin-based immunoassay. In the dual-step amplification process, detection limits for CAP in the HCR and HCR-CRISPR/Cas12a stages were significantly reduced to 55.23 pg/mL and 3.31 pg/mL, respectively. These findings introduce an effective method for developing CAP-specific scFv antibodies and also propose a multi-amplification strategy to increase immunoassay sensitivity. Additionally, theoretical studies also offer valuable guidance in CAP hapten design and genetic engineering for antibody modification.


Asunto(s)
Técnicas Biosensibles , Cloranfenicol , Humanos , Sistemas CRISPR-Cas , Células HEK293 , Hibridación de Ácido Nucleico , Fluoroinmunoensayo , Anticuerpos
17.
Biosens Bioelectron ; 252: 116139, 2024 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-38412686

RESUMEN

Micro/nanomaterials display considerable potential for increasing the sensitivity of lateral flow immunoassay (LFIA) by acting as 3D carriers for both antibodies and signals. The key to achieving high detection sensitivity depends on the probe's orientation on the material surface and its multivalent biomolecular interactions with targets. Here, we engineer Lactococcus lactis as the bacterial microcarrier (BMC) for a multivalent immunorecognition probe that was genetically programmed to display multifunctional components including a phage-screened single-chain variable fragment (scFv), an enhanced green fluorescent protein (eGFP), and a C-terminal peptidoglycan-binding domain (AcmA) anchored on BMC through the cell wall peptidoglycan. The innovative design of this biocarrier system, which incorporates a lab-on-a-chip microfluidic device, allows for the rapid and non-destructive self-assembly of the multivalent scFv-eGFP-AcmA@BMC probe, in which the 3D structure of BMC with a large peptidoglycan surface area facilitates the precisely orientated attachment and immobilization of scFv-eGFP-AcmA. This leads to a remarkable fluorescence aggregation amplification effect in LFIA, outperforming a monovalent 2D scFv-eGFP-AcmA probe for florfenicol detection. By designing a portable sensing device, we achieved an exceptionally low detection limit of 0.28 pg/mL and 0.21 pg/mL for florfenicol in lake water and milk sample, respectively. The successful microfabrication of this biocarrier holds potential to inspire innovative biohybrid designs for environment and food safety biosensing applications.


Asunto(s)
Técnicas Biosensibles , Lactococcus lactis , Tianfenicol/análogos & derivados , Animales , Antibacterianos/metabolismo , Lactococcus lactis/genética , Lactococcus lactis/química , Peptidoglicano/metabolismo , Microtecnología , Leche , Lagos , Inmunoensayo , Agua
18.
J Agric Food Chem ; 72(4): 2214-2228, 2024 Jan 31.
Artículo en Inglés | MEDLINE | ID: mdl-38237048

RESUMEN

Previously, Lactobacillus paracasei VL8, a lactobacillus strain isolated from the traditional Finnish fermented dairy product Viili, demonstrated immunomodulatory and antibacterial effects. The prebiotic mannan-oligosaccharide (MOS) further promoted its antibacterial activity and growth performance, holding promise for maintaining intestinal health. However, this has not been verified in vivo. In this study, we elucidated the process by which L. paracasei VL8 and its synbiotc combination (SYN) with MOS repair the intestinal barrier function in dextran sodium sulfate (DSS)-induced colitis mice. SYN surpasses VL8 or MOS alone in restoring goblet cells and improving the tight junction structure. Omics analysis on gut microbiota reveals SYN's ability to restore Lactobacillus spp. abundance and promote tryptophan metabolism. SYN intervention also inhibits the DSS-induced hyperactivation of the Wnt/ß-catenin pathway. Tryptophan metabolites from Lactobacillus induce intestinal organoid differentiation. Co-housing experiments confirm microbiota transferability, replicating intestinal barrier repair. In conclusion, our study highlights the potential therapeutic efficacy of the synbiotic combination of Lactobacillus paracasei VL8 and MOS in restoring the damaged intestinal barrier and offers new insights into the complex crosstalk between the gut microbiota and intestinal stem cells.


Asunto(s)
Colitis , Lacticaseibacillus paracasei , Probióticos , Simbióticos , Animales , Ratones , Sulfato de Dextran/efectos adversos , Mananos , Probióticos/farmacología , Nicho de Células Madre , Triptófano , Colitis/inducido químicamente , Colitis/genética , Colitis/terapia , Lactobacillus , Oligosacáridos , Antibacterianos/efectos adversos , Modelos Animales de Enfermedad , Ratones Endogámicos C57BL , Colon
19.
Food Chem ; 438: 138003, 2024 Apr 16.
Artículo en Inglés | MEDLINE | ID: mdl-37979258

RESUMEN

The aim of this study was to investigate the effect of different thermal processing methods on the nutritional and physicochemical qualities of Penaeus vannamei. Three different thermal processing methods, namely, drying (DS, 120 °C/40 min), steaming (SS, 100 °C/2 min), and microwaving (MS, 600 W/2 min) were used to treat the shrimps. Low-field nuclear magnetic resonance data indicated that fixed water was the main component of Penaeus vannamei. The ratio of fatty acids in MS and DS samples was more in line with the FAO/WHO recommended health requirements; The myofibrillar protein carbonyl group increased, whereas sulfhydryl content decreased after thermal processing, indicating that the proteins were oxidized by thermal processing. The magnitude of oxidation is: MS > SS > DS. Different thermal processing methods can exert great influence on color texture and nutrition to Penaeus vannamei, which can provide a theoretical knowledge for consumers to choose the appropriate processing method.


Asunto(s)
Penaeidae , Animales , Penaeidae/química , Ácidos Grasos/química , Oxidación-Reducción , Desecación , Agua
20.
Food Chem X ; 20: 101006, 2023 Dec 30.
Artículo en Inglés | MEDLINE | ID: mdl-38046180

RESUMEN

This study investigated the modification of myofibrillar protein (MP) from the razor clam through phosphorylation by using various phosphate salts, namely, sodium tripolyphosphate (STPP), sodium trimetaphosphate (STMP), sodium polyphosphate (STTP) and sodium pyrophosphate (TSPP), and their mechanisms of action for functional and gelling properties. Fourier transform infrared spectrometry (FTIR) showed that MP introduced phosphate groups during phosphorylation; these phosphates changed the secondary structure. Moreover, MP after phosphorylation led to an increase in solubility, which was more evident in the case of TSPP phosphorylation, leading to the improvement of gel properties. Therefore, TSPP was the phosphate with the best gel properties in the modification of MP, showing the highest phosphorus content, which resulted in better gelling properties owing to its relatively shorter chains. These results showed that phosphate was able to improve protein cross-linking through ion interactions and electrostatic interactions, which ultimately improved the gelling properties of the razor clam protein.

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