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Penicillium exopolysaccharide (EPS) inhibits galactose lectins and enhances immunity. However, EPS production is low and its synthesis mechanism remains unclear. Penicillium EF-2 strains with high EPS production were selected for this study, and Penicillium fermentation conditions were subsequently improved. The optimal culture conditions were 30 g/L lactose, 6 g/L yeast extract powder, 4 d seed age, 10 % inoculation amount, 3 d of secondary fermentation time, and the final EPS yield was 3.97 g/L. UHPLC-Q-TOF-MS/MS was used to explore the mechanism of EPS synthesis at the metabolic level. Optimal carbon source: lactose and optimal nitrogen source: yeast extract can provide precursors for EPS synthesis through related metabolic pathways. Moreover, regulating the energy, vitamin, and lipid metabolic pathways created favourable conditions for EPS synthesis and secretion. These findings explain the mechanism of EPS synthesis at the metabolic level and provide a theoretical basis for optimising and industrialising EPS production.
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Fermentación , Metabolómica , Penicillium , Espectrometría de Masas en Tándem , Penicillium/metabolismo , Penicillium/crecimiento & desarrollo , Metabolómica/métodos , Cromatografía Líquida de Alta Presión , Medios de Cultivo/química , Nitrógeno/metabolismoRESUMEN
Germination treatment of highland barley enhances its nutritional value while weakening the starch gel properties. This study aims to enhance the characteristics of germinated highland barley starch (GBS) by exploring the synergistic effects of two alkalis (Na2CO3 and NaHCO3) and guar gum (GG) on GBS gel properties. The combined action of alkalis and GG significantly improved the peak viscosity, setback viscosity, and hardness compared with GG alone. The highest G' and G" reached 998 and 204 Pa at 0.4% Na2CO3 addition, which were increased by nearly 44% and 50%, respectively. Fourier-transform infrared spectral analysis revealed that the alkalis strengthened interaction forces, particularly with intensified absorption peaks at 3200-3700 cm-1 and 1550-1750 cm-1. The Na2CO3 and NaHCO3 reduced the spin-spin relaxation time (T2), resulting in a dense starch gel network. This study contributes to enhancing the market application of GBS and offers innovative insights for modifying other starches.
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Hordeum , Mananos , Gomas de Plantas , Almidón , Almidón/química , Galactanos/química , Viscosidad , Geles/química , ReologíaRESUMEN
Inflammatory bowel disease is a major health problem that can lead to prolonged damage to the digestive system. This study investigated the effects of an exopolysaccharide from genistein-stimulated Monascus purpureus (G-EMP) in a mouse model of colitis to clarify its molecular mechanisms and identified its structures. G-EMP (Mw = 56.4 kDa) was primarily consisted of â 4)-α-D-Galp-(1 â, â 2,6)-α-D-Glcp-(1â and â2)-ß-D-Manp-(1 â , with one of the branches being α-D-Manp-(1 â. G-EMP intervention reduced the loss of body weight, degree of colonic damage and shortening, disease activity index scores, and histopathology scores, while restoring goblet cell production and oxidative homeostasis, repairing colonic functions, and regulating inflammatory cytokines. RNA sequencing and Western blot analysis indicated that G-EMP exerts anti-inflammatory properties by suppressing the TLR4/MAPK/NF-κB inflammatory signaling pathway. G-EMP modulated the gut microbiota by improving its diversities, elevating the relative abundances of beneficial bacteria, declining the Firmicutes/Bacteroidota value, and regulating the level of short-chain fatty acids (SCFAs). Correlation analysis demonstrated strong links between SCFAs, gut microbiota, and the inflammatory response, indicating the potential of G-EMP to prevent colitis.
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Colitis , Microbioma Gastrointestinal , Monascus , Animales , Ratones , FN-kappa B/genética , Genisteína , Receptor Toll-Like 4/genética , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Colitis/genética , Colon , Modelos Animales de Enfermedad , Sulfato de Dextran , Ratones Endogámicos C57BLRESUMEN
Polysaccharides are natural chemical compounds that are extensively employed in the food and pharmaceutical industries. They exhibit a wide range of physical and biological properties. These properties are commonly improved by using chemical and physical methods. However, with the advancement of biotechnology and increased demand for green, clean, and safe products, polysaccharide modification via microbial fermentation has gained importance in improving their physicochemical and biological activities. The physicochemical and structural characteristics, biological activity, and modification mechanisms of microbially fermented polysaccharides were reviewed and summarized in this study. Polysaccharide modifications were categorized and discussed in terms of strains and fermentation techniques. The effects of microbial fermentation on the physicochemical characteristics of polysaccharides were highlighted. The impact of modification of polysaccharides on their antioxidant, immune, hypoglycemic, and other activities, as well as probiotic digestive enhancement, were also discussed. Finally, we investigated a potential enzyme-based process for polysaccharide modification via microbial fermentation. Modification of polysaccharides via microbial fermentation has significant value and application potential.
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Exopolysaccharides from genistein-stimulated Monascus purpureus (G-EMP) exhibited immunomodulatory potential in vitro, but whether it had immune-enhancing effects in vivo and its potential mechanism are not yet known. Here, the immunomodulatory effects of G-EMP were investigated by establishing an immunosuppressed mouse model treated with cyclophosphamide (Cy). The results suggested that G-EMP effectively alleviated the signs of weight reduction and diet reduction caused by Cy, increased fecal water content and splenic index, and decreased the oxidative stress of the liver. Simultaneously, G-EMP improved Cy-induced intestinal injury by restoring villus length, increasing the number of cupped cells, upregulating the expression of mucin and tight junction proteins, and downregulating the ratio of apoptotic proteins (Bax/Bcl-2). It also boosted the levels of mouse colonic cytokines, CD4+ and CD8+ T cells. Additionally, G-EMP markedly enhanced immunomodulation via the activation of PI3K/AKT-MAPKs/NF-κB signal pathways. Furthermore, G-EMP intervention displayed a positive association with most immunological indexes by elevating the levels of short-chain fatty acids, varying gut microbiota composition, and enhancing beneficial bacteria (Lactobacillaceae, Prevotellaceae, and S24-7). These findings demonstrated that G-EMP can strengthen immunity, repair intestinal mucosal damage, regulate gut microbiota, and be a potential source of prebiotics.
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Microbioma Gastrointestinal , Monascus , Animales , Ratones , FN-kappa B , Genisteína , Fosfatidilinositol 3-Quinasas , Proteínas Proto-Oncogénicas c-akt , Linfocitos T CD8-positivos , CiclofosfamidaRESUMEN
Conventional immunochromatographic test strips (ICSs) based on gold nanoparticle (AuNP) probes offer limited sensitivity. Here, AuNPs were separately labeled with monoclonal or secondary antibodies (MAb or SAb). In addition, spherical, homogeneously dispersed, and stable selenium nanoparticles (SeNPs) were also synthesized. By optimizing the preparation parameters, two ICSs based on the dual AuNP signal amplification (Duo-ICS) or SeNPs (Se-ICS) were developed for the rapid detection of T-2 mycotoxin. The detection sensitivities of the Duo-ICS and Se-ICS assays for T-2 were 1 ng/mL and 0.25 ng/mL, respectively, which were 3-fold and 15-fold more sensitive, respectively, than a conventional ICS. Furthermore, the ICSs were applied in the detection of T-2 in cereals, which requires higher sensitivity. Our findings indicate that both ICS systems can be used for rapid, sensitive, and specific detection of T-2 toxin in cereals and potentially other sample types.
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Nanopartículas del Metal , Micotoxinas , Selenio , Oro/química , Cromatografía de Afinidad/métodos , Nanopartículas del Metal/química , Anticuerpos Monoclonales , Límite de DetecciónRESUMEN
BACKGROUND: Chinese yam fermented by Monascus, namely red mold dioscorea (RMD), has the potential of treating diseases. However, the production of citrinin limits the application of RMD. In the present study, the fermentation process of Monascus was optimized by adding genistein or luteolin to reduce citrinin yield. RESULTS: The results showed that citrinin in 25 g of Huai Shan yam was reduced by 48% and 72% without affecting the pigment yield by adding 0.2 g of luteolin or genistein, respectively, to a 250-mL conical flask after fermentation for 18 days at 28 °C, whereas the addition of luteolin increased the content of yellow pigment by 1.3-fold. Under optimal conditions, citrinin in 20 g of iron bar yam decreased by 55% and 74% after adding 0.2 g of luteolin or genistein. Luteolin also increased yellow pigment content by 1.2-fold. Ultra HPLC coupled to quadrupole time-of-flight mass spectrometry was used for the preliminary analysis of Monascus fermentation products. It was found that the amino acid types in RMD are similar to those in yams, but there are fewer polysaccharides and fatty acids. CONCLUSION: The results obtained in the present study showed that the addition of genistein or luteolin could reduce citrinin on the premise of increasing pigment yield, which laid a foundation for the better use of yams in Monascus fermentation. © 2023 Society of Chemical Industry.
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Citrinina , Dioscorea , Monascus , Fermentación , Citrinina/análisis , Dioscorea/metabolismo , Genisteína/metabolismo , Monascus/metabolismo , Luteolina/metabolismo , Pigmentos Biológicos/metabolismoRESUMEN
Exopolysaccharides are important metabolites of Monascus with healthy activities. However, the low production level limits their applications. Hence, the aim of this work was to increase the yield of exopolysaccharides (EPS) and optimize liquid fermentation by adding flavonoids. The EPS yield was optimized via both medium composition and culture conditions. The optional fermentation conditions achieved for EPS production of 7.018 g/L were 50 g/L sucrose, 3.5 g/L yeast extract, 1.0 g/L MgSO4·7H2O, 0.9 g/L KH2PO4, 1.8 g/L K2HPO4·3H2O, 1 g/L quercetin, and 2 mL/L Tween-80, with pH 5.5, inoculum size 9%, seed age 52 h, shaking speed 180 rpm, and fermentation culture 100 h, respectively. Furthermore, the addition of quercetin increased EPS production by 11.66%. The results also showed little citrinin residue in the EPS. The exopolysaccharides' composition and antioxidant capacity of quercetin-modified exopolysaccharides were then preliminarily investigated. The addition of quercetin changed the composition of the exopolysaccharides and the molecular weight (Mw). In addition, the antioxidant activity of Monascus exopolysaccharides was monitored using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS+), and -OH. Monascus exopolysaccharides have good scavenging ability of DPPH and -OH. Furthermore, quercetin increased the scavenging ABTS+ ability. Overall, these findings provide a potential rationale for the application of quercetin in improving the EPS yield.
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Monacolin K (MK), a polyketo secondary metabolic compound of the mold genus Monascus, can promote the apoptosis of malignant cancer cells, possessing potential antitumor properties. However, its mechanism of action on gliomas remains unclear. Here, we explored and investigated the potential of the monacolin K's antitumor effect on human glioma U251 cells and its possible molecular mechanism. Results showed that the application of 10 µM monacolin K inhibited the proliferation of U251 cells, with an inhibitory rate of up to 53.4%. Additionally, monacolin K induced the generation of reactive oxygen species and activated mitochondria-mediated pathways, including decreased MMP, activation of caspase3/caspase9, decreased Na+/K+-ATPase and Ca2+-ATPase activities, and disruption of the antioxidant system, resulting in the disruption of intracellular reduction-oxidation homeostasis. Monacolin K also activated MAPK and NF-κB pathways, upregulating P38 activity and downregulating JNK/ERK/P65/IκBα expression, ultimately leading to apoptosis of U251 cells. Importantly, monacolin K was not cytotoxic to normal human cells, hUC-MSCs. We concluded that monacolin K can induce apoptosis in U251 cells by triggering ROS-mediated oxidative damage and regulating MAPKs and NF-κB pathways.
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Glioma , FN-kappa B , Humanos , FN-kappa B/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Lovastatina/farmacología , Estrés Oxidativo , Glioma/metabolismo , ApoptosisRESUMEN
(1) Background: Medicinal and edible food and traditional Chinese medicine have been used to treat various diseases. However, their safety has not been thoroughly assessed. (2) Methods: An immunochromatographic test strip (ICS) was used for the first time to screen some mycotoxins, including aflatoxin B1 (AFB1), zearalenone (ZEN), and T-2 toxin, in medicinal and edible food and traditional Chinese medicine. Antibody/nano-gold particle coupling was used with the prepared ICS, and the pH, monoclonal antibody concentration, and antigen amount were optimized. The extraction sample solution was diluted 10 times with phosphate-buffered saline containing 0.5% Tween-20 and 0.05% sodium dodecyl sulfate to remove the complex matrix in medicinal and edible food. (3) Results: Under optimal conditions, the sensitivities of the developed ICS for AFB1, ZEN, and T-2 were 0.5, 5.0, and 5.0 ng/mL, respectively. Among the 30 medicinal and edible food samples tested, two samples (both of sand jujube kernels) were positive, and the results were verified by high-performance liquid chromatography and enzyme-linked immunosorbent assay and were consistent with the ICS test results. (4) Conclusions: The ICS could be used for rapid screening and simultaneous detection of mycotoxins at medicinal and edible food storage facilities.
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The exopolysaccharides (EPS) produced by the edible medicinal fungus Monascus purpureus (EMP) become the center of growing interest due to their techno-functional properties and their numerous applications in the food industries; however, the low EPS yields limit its application. In this study, the effect of genistein supplementation on the production, rheological and antioxidant properties of EPS by M. purpureus and its biosynthesis mechanism were explored. The results indicated that the addition of genistein (3 g/L) generated a 110% and 59% increase in the maximum mycelial biomass and EPS yield, respectively. The genistein supplementation group (G-EMP) had higher molar percentages of Xyl and Man, and significantly decreased molecule weight and particle size of EPS, which resulted in stronger antioxidant effect and cell growth promotion. Rheological analysis showed that both EMP and G-EMP demonstrated pseudoplastic fluid behavior and G-EMP exhibited strong gel-like elastic behavior (G' > G"). Furthermore, genistein not only facilitated the production of EPS by regulating cell membrane permeability, enhancing cellular respiratory metabolism and monosaccharide precursor synthesis pathways, and enhancing antioxidant enzyme activity to reduce oxidative stress damage, but also affected the composition of the monosaccharides by increasing enzyme activity in the underlying synthesis pathways. These findings expand the application of M. purpureus resources and provide a paradigm for future study of the structural and functional characteristics of EPS.
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Artemisia vulgaris (A. vulgaris) is a traditional Chinese medicine widely distributed in China and contains many bioactive compounds with pharmacological effects. However, the anti-inflammatory effects and mechanism of essential oil from A. vulgaris on enteritis in fish are still unclear. In this study, in order to elucidate the underlying mechanism of essential oil from A. vulgaris on zebrafish enteritis, zebrafish were used for establishing animal models to observe the histopathological changes of intestines, determine the activities of immune-related enzymes and oxidative stress indicators, and the mRNA expression of genes in MyD88/TRAF6/NF-KB signaling pathways. The results showed that different doses of A. vulgaris essential oil could effectively alleviate zebrafish enteritis in a dose- and time-dependent manner by improving the intestinal histopathological damage, decreasing the intestinal oxidative stress, repairing the intestinal immune ability, changing the expression levels of IL-1ß, IL-10 and genes in MyD88/TRAF6/NF-κB pathway. In addition, co-treatment with oxazolone and MyD88 inhibitor could alleviate the morphological damage, the induction of oxidative stress, and the levels of immune-related enzymes and the mRNA expression of genes in MyD88/TRAF6/NF-κB signaling pathway. Moreover, essential oil from A. vulgaris had more significantly therapeutic effects on enteritis of male zebrafish than that of female zebrafish. This result will clarify the therapeutic effect and anti-inflammatory mechanism of essential oil from A. vulgaris on zebrafish enteritis, and provide a theoretical basis for further research on the rationality of A. vulgaris to replace feed antibiotics.
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Artemisia , Enteritis , Aceites Volátiles , Masculino , Femenino , Animales , Pez Cebra/genética , FN-kappa B/genética , FN-kappa B/metabolismo , Artemisia/genética , Artemisia/metabolismo , Aceites Volátiles/farmacología , Factor 88 de Diferenciación Mieloide/metabolismo , Factor 6 Asociado a Receptor de TNF/genética , Factor 6 Asociado a Receptor de TNF/metabolismo , Enteritis/tratamiento farmacológico , Enteritis/veterinaria , Enteritis/genética , Estrés Oxidativo , Antiinflamatorios/farmacología , ARN Mensajero/metabolismoRESUMEN
Exopolysaccharides (EPS), metabolites of the medicinal edible fungus Monascus purpureus, have antioxidant, immunomodulatory, and anti-inflammatory effects. However, the biosynthetic mechanism of EPS from M. purpureus is still unclear, which hinders its utilization. In this study, the fermentation conditions of M. purpureus were optimized and comparative transcriptomic analysis was performed to understand the mechanisms and effects of fermentation on EPS synthesis. The optimal medium composition was 40 g/L mannose, 4 g/L yeast powder, 1 g/L MgSO4·7H2O, 0.8 g/L KH2PO4, 1.6 g/L K2HPO4·3H2O, and 2 mL/L Tween 80, and the optimal cultivation conditions were an inoculum of 7 %, culture temperature 30 °C, initial pH 6.0, and 180 rpm for 4 d. A total of 8095 unigenes were obtained, and 17 key enzymes for EPS synthesis were identified. Interestingly, 12 carbohydrate metabolism subcategories were enriched in the group with 4 days of fermentation compared to 2 days, with most of the differentially expressed genes (DEGs) being upregulated, but only nine carbohydrate metabolism subcategories were enriched with longer fermentation time, with all DEGs being downregulated. This study provides a theoretical basis for enhancing the EPS content and reveals the dynamics of EPS synthesis in M. purpureus, providing important targets for future EPS molecular modifications and gene knockdown studies.
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Monascus , Fermentación , Perfilación de la Expresión Génica , Monascus/genética , Monascus/metabolismo , Transcriptoma , Levaduras/genéticaRESUMEN
The Cyclocarya paliurus polysaccharide (CP) was chemically modified to produce sulfated derivatives (S-CP) and carboxymethylated derivatives (CM-CP). Subsequently, the antioxidant activity, cytoprotective effect and antitumor activity of these derivatives were investigated to establish the relationship between their structure and functional activity. The results found that chemical modifications resulted in remarkable variations in the chemical compositions and apparent structures of CP. S-CP with the highest amount of glucose had the strongest antioxidant capacity to scavenge DPPH⢠and HOâ¢, but CM-CP was lower than CP in terms of HO⢠scavenging. More importantly, S-CP and CM-CP more effectively protected RAW264.7 from H2O2-induced damage compared to CP by reducing the secretion of lactate dehydrogenase (LDH), intracellular reactive oxygen species (ROS) and malondialdehyde (MDA), enhancing phagocytosis and superoxide dismutase (SOD) levels, and suppressing abnormal apoptosis. Further experiments showed that the anti-apoptotic effect of S-CP and CM-CP was in intimate association with down-regulation of Caspase-9/3 activities and alleviation of cell cycle arrest in the S phase. In addition, S-CP and CM-CP decreased the cell viability of tumor cells. These findings suggest that the type of functional group plays important roles in the biological function of the derivatives and provide a theoretical basis for the development of novel natural anti-oxidants or low-toxicity anti-tumor drugs.
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Peróxido de Hidrógeno , Juglandaceae , Antioxidantes/química , Peróxido de Hidrógeno/farmacología , Juglandaceae/química , Estrés Oxidativo , Polisacáridos/química , Polisacáridos/farmacologíaRESUMEN
A polysaccharide (EMP) was prepared from Monascus purpureus 40,269 through liquid fermentation, and its immunoregulatory effect was investigated to find clues regarding its potential applicability. Structural characterization demonstrated that EMP, with a molecular weight of 83.1 kDa, consists of Xyl, Man, Gal, Ara, Rha, Glc, GalA, and GlcA at a molar ratio of 2.6: 22: 35.1: 7: 1: 29.2: 2.7: 0.6. Immunomodulatory assays involving RAW264.7 cells indicated that EMP exhibits significantly enhanced pinocytic and phagocytic capacities and promotes the secretion of reactive oxygen species, nitric oxide, and cytokines (interleukin-1ß, tumor necrosis factor-α, and interleukin-6) by activating RAW264.7 cells. Moreover, EMP is recognized by Toll-like receptor 4 (TLR-4) and has an immunomodulatory effect by activating the mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) signaling pathways in RAW264.7 cells. Collectively, these results demonstrate that EMP could be used as a functional food for immunological reagents.
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Sistema de Señalización de MAP Quinasas , Monascus , FN-kappa B , Polisacáridos , Receptores de Laminina , Receptor Toll-Like 4 , Animales , Fermentación , Factores Inmunológicos/química , Factores Inmunológicos/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Ratones , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Monascus/química , Monascus/metabolismo , FN-kappa B/metabolismo , Polisacáridos/química , Polisacáridos/farmacología , Células RAW 264.7 , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/metabolismoRESUMEN
Manipulating the structures, physicochemical properties, and monosaccharide compositions of exopolysaccharides (EPS) isolated from microorganisms has been reported to enhance their biological activities. Hence, the aim of this work was to examine the effects of genistein addition during fermentation on the amount, physicochemical properties, and immunomodulatory activity of EPS secreted by M. purpureus. Results showed that genistein addition significantly increased M. purpureus biomass and EPS yield to 2.42 g L-1 and 6.08 g L-1, respectively, and affected the physicochemical properties and structures of EPS. Furthermore, EPS produced by genistein-treated M. purpureus (G-EMP) improved the immunomodulatory activity of RAW264.7 macrophages by increasing the secretion of nitric oxide and cytokines. Moreover, phospho-Jun N-terminal kinase (p-JNK), phospho-extracellular regulated protein kinase (p-ERK), phospho-p38 (p-p38) mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) phospho-p65 (p65) proteins were remarkably upregulated by G-EMP stimulation, blocking Toll-like receptor 4 (TLR4) that dramatically reduced the pinocytic and phagocytic capacities. Overall, these findings provide potential rationales for the application of genistein in improving the EPS yield of M. purpureus.
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Genisteína/farmacología , Factores Inmunológicos/farmacología , Monascus , Polisacáridos/biosíntesis , Animales , Ratones , Células RAW 264.7/efectos de los fármacosRESUMEN
INTRODUCTION: Plants containing aristolochic acid and its derivatives are nephrotoxic, mutagenic, and carcinogenic to humans; chronic diet poisoning caused by the aristolochic acid is the cause of endemic (Balkan) nephropathy and related cancers. OBJECTIVE: To develop a colloidal gold immunochromatographic test strip (ICS) based on the competitive format for the rapid detection of aristolochic acid A (AA-A) in herbal medicinal materials. MATERIALS AND METHODS: For the ICS based on gold nanoparticles (AuNPs), the antigen [AA-A-bovine serum albumin (BSA)], and goat anti-mouse IgG were drawn on the nitrocellulose membrane as the test line (T line) and the control line (C line), respectively. Monoclonal antibody (MAb)-AuNP conjugates were sprayed onto the conjugate pad. The sensitivity of the ICS was 6 ng/mL, and the test was completed in 10 min. The analysis of AA-A in traditional Chinese medicine samples showed that the ICS results were in good agreement with those obtained by high-performance liquid chromatography methods. CONCLUSION: These results demonstrated that the ICS test could be used as a reliable, rapid, cost-effective, and convenient qualitative tool for on-site screening techniques to detect AA-A in herbal medicinal materials without any special instrumentation.
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Ácidos Aristolóquicos , Nanopartículas del Metal , Animales , Oro , Oro Coloide/química , Ratones , Sensibilidad y EspecificidadRESUMEN
Monascus, which is traditionally used in various Asian industries, produces several secondary metabolites during the fermentation process, including citrinin, a toxin whose impact limits the development of the Monascus industry. We have previously found that the addition of 2.0 g/L genistein to Monascus medium reduces citrinin production by approximately 80%. Here, we explored the molecular mechanisms whereby genistein affects citrinin production. We sequenced the Monascus genome and performed transcriptome analysis on genistein-treated and -untreated groups. Comparison between the two groups showed 378 downregulated and 564 upregulated genes. Among the latter, we further examined the genes related to citrinin biosynthesis and quantified them using quantitative real-time polymerase chain reaction (qRT-PCR). Genes orf5, pksCT, orf3, orf1, orf6, and ctnE were significantly downregulated, demonstrating that genistein addition indeed affects citrinin synthesis. Our results may lay the groundwork for substantial improvements in the Monascus fermentation industry.