استكشف المجموعة الواسعة من العناوين المتاحة.

Biosynthetic enzymes evolutionarily gain novel functions, thereby expanding the structural diversity of natural products to the benefit of host organisms. Diels-Alderases (DAs), functionally unique enzymes catalysing [4 + 2] cycloaddition reactions, have received considerable research interest. However, their evolutionary mechanisms remain obscure. Here, we investigate the evolutionary origins of the intermolecular DAs in the biosynthesis of Moraceae plant-derived Diels-Alder-type secondary metabolites. Our findings suggest that these DAs have evolved from an ancestor functioning as a flavin adenine dinucleotide (FAD)-dependent oxidocyclase (OC), which catalyses the oxidative cyclisation reactions of isoprenoid-substituted phenolic compounds. Through crystal structure determination, computational calculations, and site-directed mutagenesis experiments, we identified several critical substitutions, including S348L, A357L, D389E and H418R that alter the substrate-binding mode and enable the OCs to gain intermolecular DA activity during evolution. This work provides mechanistic insights into the evolutionary rationale of DAs and paves the way for mining and engineering new DAs from other protein families.

The Diels-Alder reaction is one of the most powerful and widely used methods in synthetic chemistry for the stereospecific construction of carbon-carbon bonds. Despite the importance of Diels-Alder reactions in the biosynthesis of numerous secondary metabolites, no naturally occurring stand-alone Diels-Alderase has been demonstrated to catalyse intermolecular Diels-Alder transformations. Here we report a flavin adenine dinucleotide-dependent enzyme, Morus alba Diels-Alderase (MaDA), from Morus cell cultures, that catalyses an intermolecular [4+2] cycloaddition to produce the natural isoprenylated flavonoid chalcomoracin with a high efficiency and enantioselectivity. Density functional theory calculations and preliminary measurements of the kinetic isotope effects establish a concerted but asynchronous pericyclic pathway. Structure-guided mutagenesis and docking studies demonstrate the interactions of MaDA with the diene and dienophile to catalyse the [4+2] cycloaddition. MaDA exhibits a substrate promiscuity towards both dienes and dienophiles, which enables the expedient syntheses of structurally diverse natural products. We also report a biosynthetic intermediate probe (BIP)-based target identification strategy used to discover MaDA.

Neochlorogenic acid (nCGA) is a phenolic compound isolated from mulberry leaf ( ), which possesses multiple pharmacological activities containing antioxidant and anti-inflammatory effects. However, the role of nCGA in the treatment of acute pneumonia and the underlying molecular mechanism are still unclear. Hence, the aim of study is to investigate the anti-inflammatory properties of nCGA on LPS-stimulated inflammation in A549 cells. In the present study, results reported that nCGA without cytotoxicity significantly reduced the production of TNF-α, IL-6, and NO, and further suppressed the proteins of iNOS, COX2, TNF-α, IL-6 expression. Furthermore, nCGA also inhibited NF-κB activation and blocked MAPKs signaling pathway phosphorylation. In addition, we found nCGA significantly increased the expression of HO-1 via activating the AMPK/Nrf2 signaling pathway to attenuate the inflammatory response, whereas this protective effect of nCGA was reversed by pre-treatment with compound C (C.C, an AMPK inhibitor). Therefore, all these results indicated that nCGA might act as a natural anti-inflammatory agent for the treatment of acute pneumonia.

The antidiabetic and antioxidant activities of the ethyl acetate-soluble extract (MFE) of mulberry fruit (Morus alba L.) were investigated. In vitro, MFE showed potent α-glucosidase inhibitory activity and radical-scavenging activities against DPPH and superoxide anion radicals. In vivo, MFE could significantly decrease fasting blood glucose (FBG) and glycosylated serum protein (GSP), and increase antioxidant enzymatic activities (SOD, CAT, GSH-Px) in streptozotocin (STZ)-induced diabetic mice. Bioactivity-guided fractionation of the MFE led to the isolation of 25 phenolic compounds, and their structures were identified on the basis of MS and NMR data. All the 25 compounds were isolated from mulberry fruit for the first time. Also, the α-glucosidase inhibitory activity and antioxidant activity of the phenolics were evaluated. Potent α-glucosidase inhibitory and radical-scavenging activities of these phenolics suggested that they may be partially responsible for the antidiabetic and antioxidant activities of mulberry fruit.

Herein, we report the synthesis of silver nanoparticles (AgNPs) by a green route using the aqueous leaf extract of Morus indica L. V1. The synthesized AgNPs exhibited maximum UV-Vis absorbance at 460 nm due to surface plasmon resonance. The average diameter (~54 nm) of AgNPs was measured from HR-TEM analysis. EDX spectra also supported the formation of AgNPs, and negative zeta potential value (-14 mV) suggested its stability. Moreover, a shift in the carbonyl stretching (from 1639 cm to 1630 cm ) was noted in the FT-IR spectra of leaf extract after AgNPs synthesis which confirm the role of natural products present in leaves for the conversion of silver ions to AgNPs. The four bright circular rings (111), (200), (220) and (311) observed in the selected area electron diffraction pattern are the characteristic reflections of face centered cubic crystalline silver. LC-MS/MS study revealed the presence of phytochemicals in the leaf extract which is responsible for the reduction of silver ions. MTT assay was performed to investigate the cytotoxicity of AgNPs against two human cell lines, namely HepG2 and WRL-68. The antibacterial study revealed that MIC value of the synthesized AgNPs was 80 µg/ml against Escherichia coli K12 and Staphylococcus aureus (MTCC 96). Finally, the synthesized AgNPs at 10 µg/ml dosages showed beneficial effects on the survivability, body weights of the Bombyx mori L. larvae, pupae, cocoons and shells weights via enhancing the feed efficacy.

L. (white mulberry) has been commonly used as a functional food and for medicinal purposes. Hence, the aim of the study was to compare the phenolic profile of white mulberry commercial samples in relation to their antioxidant potential and acetylcholinesterase (AChE) inhibitory activity. It is of interest to determine whether herbal products originating from different commercial sources differ in their phenolic profiles. For this purpose, a simple and rapid high-performance liquid chromatography (HPLC) method was used for the separation and determination of ten major phenolic compounds. Total phenolic (TPC), total flavonoid (TFC), and total phenolic acid contents (TPAC), as well as l(+)-ascorbic acid (ASA) contents, were determined. The antioxidant potential was assessed by DPPH (2,2-diphenyl-1-picrylhydrazyl radical) scavenging activity and ferric-reducing/antioxidant power (FRAP) assay, while the AChE inhibitory activity was determined by the Ellman assay for water extracts. The study revealed that excluding two herbal products containing fruits and a sample containing leaves of white mulberry, yerba mate and lemon, the remaining samples were generally consistent in terms of phenolic composition as well as antioxidant potential and AChE inhibitory activity. This reflects the health-promoting properties of the samples under study. Moreover, the results showed that the water extracts of white mulberry were richer in phenolic compounds and presented higher antioxidant activity than the hydromethanolic extracts. However, the water extracts showed low inhibitory activity against AChE. Moreover, the correlation analysis indicated a high positive relationship between phenolic composition and antioxidant activity in extracts of white mulberry. Overall, the obtained results may be useful in the evaluation of new dietary supplements and food products. The water extracts of white mulberry could be used for antioxidant purposes, while the hydromethanolic extracts could be incorporated in antioxidant formulations.

Type II diabetes mellitus (T2DM) is the most common form of diabetes and has become a major health problem across the world. The root bark of L. is widely used in Traditional Chinese Medicine for treatment and management of diabetes. The aim of the present study was to evaluate the enzyme inhibitory potentials of three principle components, mulberrofuran G ( ), albanol B ( ), and kuwanon G ( ) in root bark against diabetes, establish their enzyme kinetics, carry out a molecular docking simulation, and demonstrate the glucose uptake activity in insulin-resistant HepG2 cells. Compounds ⁻ showed potent mixed-type enzyme inhibition against protein tyrosine phosphatase 1B (PTP1B) and α-glucosidase. In particular, molecular docking simulations of ⁻ demonstrated negative binding energies in both enzymes. Moreover, ⁻ were non-toxic up to 5 µM concentration in HepG2 cells and enhanced glucose uptake significantly and decreased PTP1B expression in a dose-dependent manner in insulin-resistant HepG2 cells. Our overall results depict ⁻ from root bark as dual inhibitors of PTP1B and α-glucosidase enzymes, as well as insulin sensitizers. These active constituents in may potentially be utilized as an effective treatment for T2DM.

Guangdong, Guangxi and Chongqing are emerging sericulture areas in China where the production of mulberry leaves is huge. In order to identity high quality mulberry leaves that are suitable for healthy products to expand planting, 24 samples from three regions (Guangdong, Guangxi, Chongqing) in the south of China were quantified for two alkaloids (1-deoxynojirimycin and fagomine) and five phenols (chlorogenic acid, rutin, isoquercitrin, etc.) using high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). Additionally, the total phenolic and total flavonoid contents, antioxidant and glycosidase inhibitory activities (hypoglycemic activity) were tested using different assays (DPPH, ABTS, FRAP) to comprehensively evaluate the quality of the mulberry leaves. The contents of DNJ and fagomine ranged from 0.401±0.003 to 5.309±0.036 mg/g and from 0.279±0.031 to 2.300±0.060 mg/g, respectively. The main phenolic constituents were chlorogenic acid, rutin and isoquercitrin, with chlorogenic acid present in the highest concentrations, ranging from 3.104±0.191 to 10.050±0.143 mg/g. The antioxidant activity exhibited a tendency as follows: Guangxi > Guangdong > Chongqing, except for two samples from Chongqing, which showed the highest antioxidant activity. Based on our study, mulberry leaves from Guangdong and Guangxi could be future sources of natural hypoglycemic and antioxidant products.

Natural stilbenes have unique physiological effects, such as anti-senile dementia, anti-cancer, anti-bacterial, lowering blood lipid, and other important biological functions, which have attracted great attention from scholars in recent years. In this study, two stilbene compounds, resveratrol (RES) and polydatin (PD), were isolated from Mulberry ( L.), and their antioxidant activity and mechanism were investigated. The results showed that the contents of RES and PD in mulberry roots were 32.45 and 3.15 μg/g, respectively, significantly higher than those in mulberry fruits (0.48 and 0.0020 μg/g) and mulberry branches (5.70 and 0.33 μg/g). Both RES and PD showed high antioxidant potential by DPPH, ABTS free-scavenging methods, and ORAC assay, and provided protection against oxidative damage in HepG2 cells by increased catalase (CAT) activity, superoxide dismutase (SOD) activity, and Glutathione (GSH) content, and decreasing generation of reactive oxygen species (ROS), lactate dehydrogenase (LDH) level, and malondialdehyde (MDA) content. Therefore, RES and PD treatment could be effective for attenuating AAPH-induced oxidative stress in HepG2 cells. This study will promote the development and application of stilbene compounds. Furthermore, the RES and PD could be used as antioxidant supplements in functional foods, cosmetics, or pharmaceuticals, contributing to health improvement.

In this study, leaves of three indigenous varieties of Mulberry namely, Morus alba L., Morus nigra L. and Morus rubra L. were investigated for their antioxidant potential and their proximate composition was determined. The yields of 80% methanolic extracts ranged between 8.28-13.89%. The contents of total phenolics (TPC), total flavonoids (TFC) and ascorbic acid (AA) ranged between 16.21-24.37 mg gallic acid equivalent (GAE)/g, 26.41-31.28 mg rutin equivalent (RE)/g and 0.97-1.49 mg/g, respectively. The antioxidant activity of leaf extracts was evaluated by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH(•)) radical scavenging actity, 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid (ABTS(•+)) radical cation scavenging capacity and ferric ion reducing power and values ranged between 1.89-2.12, 6.12-9.89 and 0.56-0.97 mM Trolox equivalent/g of dried leaves, respectively. The investigated features reveal good nutritive and antioxidant attributes of all the varieties with mutually significant differences.