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Coprinus comatus is an edible and medicinal fungus. In this study, the antioxidant activity of the fermentation product of C. comatus was investigated through optimization of fermentation process. The results indicated that the fermentation product of C. comatus had obvious scavenging ability for 2,2'-Azino-bis(3-ethylbenzothiazoline)-6-sulphonic acid (ABTS) free radical. The EC50 of the n-butanol extract from the fermentation product on ABTS·+ was 0.65 ± 0.02 mg/mL. On this basis, the liquid fermentation conditions of C. comatus were optimized through single factor and response surface optimization experiments according to the scavenging ability of ABTS·+ to improve the antioxidant capacity of the fermentation product. The results showed that when the 14% of C. comatus was fermented in a culture medium with a C/N ratio of 48:1 for 6 days, the ABTS·+ scavenging ability was the strongest, and the EC50 of n-butanol extract was 0.57 ± 0.01 mg/mL, which was 12.31% higher than the initial activity. This study laid the foundation for the development of C. comatus. [ABSTRACT FROM AUTHOR]

A high-speed counter-current chromatographic (HSCCC) method using ethyl acetate–water as solvent system was established to separate isoquercetin from Toona sinensis. In an HSCCC single separation, the sample sizes of ethanol extract were optimized from 203 to 1200 mg. The results showed that the yield of the target compound increased from 4 to 26 mg, and the corresponding purity decreased from 93.30 to 81.82%. To further improve the yield and purity, liquid–liquid-refining extraction was introduced to pretreat the ethanol extract and enrich the target compound. The ethanol extract was extracted with n -hexane-ethyl acetate-method-water (1:5:1:5, v/v) and ethyl acetate-water in turn to remove the low-polarity and high-polarity impurities and obtain the crude sample. Under the similar conditions, 85.25 mg of the target compound with the purity of 95.12% was separated from 240 mg of the crude sample. Subsequently, a consecutive HSCCC was developed to obtain 257 mg of the target compound from 720 mg of crude sample, which was equivalent to 14.4 g of ethanol extract. This method improved the purity of the target compound, but more importantly, the sample size can reach 12 times of the maximum sample size of the ethanol extract in a single run. [ABSTRACT FROM AUTHOR]

Ellagic acid is one of the most representative natural antioxidants, and is rich in pomegranate peel. In this study, a consecutive countercurrent chromatographic (CCC) separation method was established to improve the preparative efficiency of ellagic acid from pomegranate peel. By optimizing the solvent system, sample size and flow rate, 280 mg of ellagic acid was obtained from 5 g of crude sample from pomegranate peel by CCC after six consecutive injections. Moreover, the values of EC50 for ellagic acid in scavenging ABTS·+ and DPPH· were 4.59 ± 0.07 and 10.54 ± 0.07 μg/ml, respectively, indicating a strong antioxidant activity. This study not only established a high‐throughput method for the preparation of ellagic acid, but also provided a successful example for the development of and research on other natural antioxidants. [ABSTRACT FROM AUTHOR]

Agaricus bisporus is one of the most widely cultivated edible mushrooms in the world. The chemical components of A. bisporus have a wide range of biological activities. In order to deeply understand the antioxidant properties of A. bisporus, this study conducted an investigation on the components of A. bisporus fermentation. Through the single factor experiment and response surface optimization, it was found that when the C/N ratio was 45:1, the inoculum concentration was 10%, and the fermentation time was 7 d, the n-butanol extract of the fermentation product had the strongest scavenging capacity for free radical generated through 2,2'-azino-bis(3-ethylbenzothiazoline)-6-sulphonic acid (ABTS·+). The concentration for 50% of the maximal effect (EC50) was 0.33 ± 0.01 mg/mL. Moreover, in order to identify the two main components, the elution–extrusion counter-current chromatography (EECCC) was employed for separation, where 5,5'-oxy-dimethyl-bis(2-furfuraldehyde) and 5-(butoxymethyl) furfural were obtained. The antioxidant activity of 5,5'-oxy-dimethyl-bis(2-furfuraldehyde) (EC50 = 0.26 ± 0.01 mg/mL) was superior to that of 5-butylmethyl furfural (EC50 = 1.52 ± 0.02 mg/mL), indicating that 5,5'-oxy-dimethyl-bis(2-furfuraldehyde) was the main antioxidant in the fermentation products. The thermodynamic parameters and frontier molecular orbitals of 5,5'-oxy-dimethyl-bis (2-furanaldehyde) was evaluated by density functional theory (DFT). The result indicated 5,5'-oxy-dimethyl-bis(2-furanaldehyde) scavenged free radicals in polar media through single electron transfer followed by proton transfer (SET-PT). [ABSTRACT FROM AUTHOR]

Morus alba leaves are a natural product with great antidiabetic potential. However, the therapeutic efficacy of natural products is usually achieved through the interaction of active compounds with specific targets. Among them, active compounds with multi‐target therapeutic functions are more effective than single‐target enzymes. In this study, a bienzyme system was constructed by co‐immobilizing α‐amylase and α‐glucosidase onto Fe3O4 for affinity screening of dual‐target active components in the complex extract from M. alba leaves. As a result, a potential active compound was selectively screened by ligand fishing, separated by high‐speed countercurrent chromatography using a solvent system of ethyl acetate–n‐butanol–water (3:2:5, v/v), and identified as rutin. In addition, the result of molecular docking showed that rutin could interact with the active center of α‐amylase and α‐glucosidase through multiple hydrogen bonds, van der Waals forces, etc. to play an inhibitory role. These results demonstrate the effectiveness of the polydopamine magnetically immobilized bienzyme system for dual‐target affinity screening of active substances. This study not only reveals the chemical basis of the antidiabetic activity of M. alba leaves from a dual‐target perspective, but also promotes the progress of multitarget affinity screening. [ABSTRACT FROM AUTHOR]

Diabetes is a chronic metabolic disease caused by insufficient insulin secretion and insulin resistance. Natural product is one of the most important resources for anti-diabetic drug. However, due to the extremely complex composition, this research is facing great challenges. After the advent of ligand fishing technology based on enzyme immobilization, the efficiency of screening anti-diabetic components has been greatly improved. In order to provide critical knowledge for future research in this field, the application progress of immobilized enzyme in screening anti-diabetic components from complex natural extracts in recent years was reviewed comprehensively, including novel preparation technologies and strategies of immobilized enzyme and its outstanding application prospect in many aspects. The basic principles and preparation steps of immobilized enzyme were briefly described, including entrapment, physical adsorption, covalent binding, affinity immobilization, multienzyme system and carrier-free immobilization. New formatted immobilized enzymes with different carriers, hollow fibers, magnetic materials, microreactors, metal organic frameworks, etc., were widely used to screen anti-diabetic compositions from various natural products, such as Ginkgo biloba, Morus alba, lotus leaves, Pueraria lobata, Prunella vulgaris, and Magnolia cortex. Furthermore, the challenges and future prospects in this field were put forward in this review. [ABSTRACT FROM AUTHOR]