Mogroside IIIE Research Articles

Metabolism of Two Mogroside Isomers, Mogroside III and Mogroside IIIE, in Normal and Drug-Metabolizing Enzyme-Induced Rats.

Mogrosides III (1) and IIIE (2) are two important bioactive cucurbitane-type triterpenoid triglycosides in the edible fruits of Siraitia grosvenorii (Swingle), which are isomers and have only a minor difference in their structures. To clarify the effects of structural difference and drug-metabolizing-enzyme induction on their metabolism in vivo, their metabolites in normal rats and drug-metabolizing-enzyme-induced rats were tentatively identified and semiquantified by using the HPLC-DAD-ESI-IT-TOF-MSn technique. Totally, 76, 78, 96, and 121 metabolites of mogrosides were identified in the NIII (normal rats orally administered with mogroside III), NIIIE (normal rats orally administered with mogroside IIIE), EIII (drug-metabolizing-enzyme-induced rats orally administered with mogroside III), and EIIIE (drug-metabolizing-enzyme-induced rats orally administered with mogroside IIIE) groups, respectively. The metabolite differences among these groups indicated that their minor structural differences are responsible for the significant differences in their metabolites, and the induction of drug-metabolizing enzymes significantly increased the number of their metabolites. These findings would improve our understanding of the in vivo processes of mogrosides III and IIIE as well as their interactions with other food bioactive components or drugs.

Inhibition of Mogroside IIIE on isoproterenol-induced myocardial fibrosis through the TLR4/MyD88/NF-κB signaling pathway.

To investigate the effect of mogroside IIIE (MGIIIE) on isoproterenol (ISO)-induced myocardial fibrosis and explore its possible mechanisms. Forty C57BL/6 male mice (6-8 weeks) were randomly divided into a control group (n=10), model group (n=10), low MGIIIE dose group (n=10), and high MGIIIE dose group (n=10). Myocardial fibrosis was established by subcutaneous ISO injection. After 2 weeks of continuous gastric administration of MGIIIE, the cardiac structure was evaluated by echocardiography. Myocardial inflammation and fibrosis were evaluated by histology examination. Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), p-IκBα, p-NF-κB, transforming growth factor β1 (TGF-β1), and α-smooth muscle actin (α-SMA) expression were detected by western blot. Inflammatory cytokines (IL-1β, IL-6, and TNF-α) in the serum were examined by ELISA. In the in vitro study, Ang II (1 μmol/l) was used to stimulate the fibroblasts, then inflammation and fibrosis index were detected. MGIIIE inhibited inflammation and fibrosis and down-regulated TLR4, MyD88, TGF-β1, and α-SMA expression in the myocardium. In the in vitro study, MGIIIE ameliorates the deposition of Col Ш and Col I and decreases the release of inflammatory cytokines. MGIIIE increased p-IκBα and reduced p-NF-κB expression both in vivo and in vitro. MGIIIE plays a role in anti-myocardial fibrosis, by inhibiting TLR4/MyD88/NF-κB signaling expression, and decreasing inflammatory cytokine release. MGIIIE may represent a novel therapeutic strategy for treating cardiac fibrosis.

Enzymatic hydrolyzation of mogrosides in Luo Han Guo extract by NKA-adsorbed snailase improves its sensory profile

Extracts of Siraitia grosvenorii (Swingle), in Chinese known as Luo Han Guo (LHG), is authorized for use as a natural sweetener. LHG is rich in mogroside V that contains five glucoses, but also contains mogroside IIIE and analogues with fewer than three glucose units that cause an unpleasant aftertaste, limiting the use of the extract. Snailase was applied here to convert mogroside V in LHG extract in favor of siamenoside I formation, the sweetest mogroside with a taste similar to sucrose. For application, snailase was immobilized by adsorption to NKA (a macroporous resin), resulting in 10.9 U per g of adsorbed protein. Reuse of the NKA-adsorbed snailase was demonstrated for four cycles, and a continuous production of improved LHG extract at a 0.5 L scale had a productivity of 68.4 g/(L⋅day). The resulting product containing over 50% siamenoside I displayed an improved taste profile with satisfying safety toward HEK293T cells.

Fruit derived callus and cell suspension culture as promising alternative sources for mogrosides production in Siraitia grosvenorii (Swingle) C. Jeffrey: a zero-caloric natural sweetener

The fruit of Siraitia grosvenorii (Luo Han Guo) is commercially consumed for its zero-calorie natural sweetener compounds known as mogrosides. The present work aimed to establish a cell culture platform from the fruit mesocarp and provide an alternative source for mogrosides production in S. grosvenorii. Bavistin at 1.0% with mercuric chloride at 0.1% were effective concentrations for explants sterilization. The maximum frequency of callus induction was observed in mesocarp (95.78%) and stem (97%) explants cultured on Murashige and Skoog (MS) medium supplemented with thidiazuron (0.5 mg/L) and picloram (2.5 mg/L). The maximum proliferation response of suspended cells in suspension culture was observed in MS medium fortified with thidiazuron (0.3 mg/L) and indole-3-butyric acid (0.3 mg/L). Mogroside V (2.96 mg/g DW), 11-oxo-mogroside V (0.66 mg/g DW), siamenoside I (0.26 mg/g DW), and mogroside IIIE (0.08 mg/g DW) were quantified in mesocarp callus using UPLC–PDA–ESI-Q-TOF-MS technique. A comparable content of mogroside V (1.76 mg/g DW) and 11-oxo-mogroside V (0.68 mg/g DW) was also quantified in cell suspension derived from mesocarp callus with enhanced cell biomass (FW; 184.58 g/L). Therefore, the in vitro culture derived from fruit mesocarp was shown to be a potential source for mogrosides production.

Engineering of a UDP-Glycosyltransferase for the Efficient Whole-Cell Biosynthesis of Siamenoside I in Escherichia coli.

The combination of the insufficient availability and the complex structure of siamenoside I (SI), the sweetest glucoside isolated from Siraitia grosvenorii to date, limited its use as a natural sweetener. To solve this problem, an improved biocatalyst, UGT-M2, was semi-rationally created by engineering the uridine diphosphate glycosyltransferase UGT94-289-2 from S. grosvenorii for the monoglucosylation of mogroside IIIE (MG IIIE) to SI. Subsequently, an engineered Escherichia coli cell was constructed, which combined UGT-M2 with a UDP-glucose regeneration system to circumvent the need for expensive UDP-glucose to produce SI. After optimization, high-purity SI (>96.4%) was efficiently prepared from MG IIIE at a 1 L scale with a productivity of 29.78 g/(L day) and a molar yield of 76.5% and without using exogenous UDP-glucose. This study not only developed a whole-cell approach for the preparation of SI but also provided an alternative glycosyltransferase variant for SI biosynthesis with synthetic biology in the future.

Selective enzymatic α-1,6- monoglucosylation of mogroside IIIE for the bio-creation of α-siamenoside I, a potential high-intensity sweetener

A new compound, α-siamenoside I (α-SI), with a glucose unit selectively bound to the 6-hydroxyl group of the 24-O-β-glucosyl moiety of mogroside IIIE by α-1,6-glucosidic bond, was bio-created by two screened cyclodextrin glycosyltransferases with a maximum yield of 59.3%. Compared to mogroside IIIE, α-SI showed a significantly increased sweetness intensity (508 times sweeter than 5% sucrose), which is superior to siamenoside I (SI), the sweetest triterpenoid saponin isolated from Siraitia grosvenorii to date. Sensory evaluation showed that the taste quality of α-SI also was obviously better than mogroside IIIE. In addition to α-SI possessing a good stability similar to that of SI, it also did not cause a significant decrease in cell viability at a concentration of 200 μg/mL and had a negative influence on islets function at 1 μM. All of these preliminarily results pave the way for promoting α-SI as a potential low-calorie sweetener.

Comparative In vitro metabolism of purified mogrosides derived from monk fruit extracts

Mogrosides are the primary components responsible for the sweet taste of Monk fruit which is derived from Siraitia grosvenorii (Swingle), a herbaceous plant native to southern China. Many mogrosides have been identified from Monk fruit extract, but the major sweetness component of Monk fruit by mass is mogroside V, comprising up to 0.5% of the dried fruit weight. Recent pharmacokinetic studies indicate that the parent mogrosides undergo minimal systemic absorption following ingestion and hydrolysis by digestive enzymes and/or intestinal flora and are excreted as mogrol (i.e., the aglycone) and its mono- and diglucosides. The objective of this study was to demonstrate whether individual mogrosides, are metabolized to a common and terminal deglycosylated metabolite, mogrol. An in vitro assay was conducted with pooled human male and female intestinal fecal homogenates (HFH) using mogrosides IIIe, mogroside V, siamenoside I, and isomogroside V at two concentrations over a 48 h period. The results show that various mogrosides that differ in the linkages and number of glucose units attached to a common cucurbitane backbone, share a common metabolic fate, and are metabolized within 24 h to mogrol. Aside from an apparent difference in the initial rate of deglycosylation between mogrosides at higher concentrations, no apparent difference in the rate of deglycosylation was observed between the male and female HFH. Given the similar structures of these mogrosides and a shared metabolic fate to mogrol, the study provides support for a reasonably conservative approach to assess safety based on bridging safety data from an individual mogroside (i.e., Mogroside V) to other mogrosides, and the establishment of a group Acceptable Daily Intake (ADI), rather than individual ADI's for mogrosides.

Mogroside IIIE Alleviates High Glucose-Induced Inflammation, Oxidative Stress and Apoptosis of Podocytes by the Activation of AMPK/SIRT1 Signaling Pathway.

BackgroundDiabetic nephropathy (DN) is the leading cause of impaired renal function. The purpose of this study was to investigate the effects of Mogroside IIIE (MG IIIE), a cucurbitane-type compound isolated from Siraitia grosvenorii, in high glucose (HG)-induced podocytes and the possible mechanisms.MethodsMPC-5 cells were cultured under normal glucose or HG conditions. After treatment with MG IIIE, cell viability was examined using a cell counting kit-8 assay. The contents of inflammatory factors and oxidative stress-related markers were determined using the corresponding kits. Additionally, apoptosis of MPC-5 cells was determined using flow cytometry assay and the levels of apoptosis-associated proteins were evaluated by Western blot analysis. Moreover, the expression of proteins in AMPK/SIRT1 signaling was tested and the compound C, an AMPK inhibitor, was used to study whether the effects of MG IIIE on HG-induced MPC-5 cells were mediated by activation of the AMPK/SIRT1 signaling pathway.ResultsMG IIIE elevated the cell viability of HG-induced MPC-5 cells, reduced the concentrations of inflammatory cytokines and decreased the levels of oxidative stress-related markers. What’s more, the apoptosis of podocytes induced by HG was inhibited after MG IIIE intervention, accompanied by the upregulated expression of Bcl-2 and downregulated expression of Bax, cleaved caspase-3 and cleaved caspase-9. It was also found that MG IIIE could activate the AMPK/SIRT1 signaling, but compound C inhibited this pathway and reversed the inhibitory effects of MG IIIE on inflammation, oxidative stress and apoptosis in HG-stimulated podocytes.ConclusionMG IIIE can alleviate HG-induced inflammation and oxidative stress of podocytes by the activation of AMPK-SIRT1 signaling.

Analysis of Mogrosides in Siraitia grosvenorii Fruits at Different Stages of Maturity

Monk fruit extract has been approved as a natural sweetener by many countries. Its major sweet components, mogrosides, display different sweet intensities and profiles. Therefore, it is important to understand the change of mogroside contents in Siraitia grosvenorii at different maturity stages. In this study, monk fruit cultivars were collected from 4 locations in GuangXi, GuiZhou, and HuNan, at different times. Mogroside IIe, mogroside III, mogroside IIIe, mogroside IV, mogroside V, isomogroside V, and siamenoside I in each sample were quantified using high performance liquid chromatography coupled to triple quadrupole mass spectrometer (LC-MS-MS). As a result, mogroside IIe was the major component at the early maturity stage. It is converted to mogroside III from 15 to 45 days, then continued the glycosylation rapidly to yield mogroside V which kept predominant after 60 days. Highly glycosylated mogrosides, such as mogroside V and siamenoside I, which provide a better taste profile, accumulated and stabilized from 75 to 90 days. It is recommended to harvest the fruit after 75 days of pollination.

Immobilization of enzymes into nanofiber membranes via electrospinning and its application in natural sweetener production

Non-caloric artificial sweeteners (NAS) are currently the world's most widely used food additives, which are considered safe and contributed to good health. But recent research shows that there are still controversies. Therefore, natural sweeteners had gained much attention as NAS substitutes. Recently, the fastest growing market of natural sweetener is the sweeteners made from monk fruit, because its extracts are highest in sweetness among various natural sweeteners and parts of the sweetness compounds have significant anti-diabetic activity. However, one of the components in monk fruit extracts called Siamenoside I (S I), which found to be the most sweetness compound and taste most closely to sucrose, are relatively low in contents. In addition, the Mogroside III E (MG III E), which is lower in sweetness compared to S I but with significant anti-diabetic properties, are also low in contents in the natural monk fruit extracts. As the results, the developing of methods to enrich the contents of S I and MG III E in monk fruit extracts and related products are the goals for the current natural sweetener manufacturers. The bitter tastes and flavors of saponins containing functional foods and related beverages are still the issues to be addressed. For example, the modification of glycoside linkage patterns of saponins including oleuropeins, ginsenosides, and momordicosides could achieve the debittering purpose and meet the consumers' preferences. Therefore, the goals of this research are to immobilize specific enzymes using electrospinning. Together with the high throughput microreactor, and the continuous packed-bed column, we suppose that we could obtain a fair amount of S I and MG III E through enzyme transformation of MG V and improve the taste of monk fruit extracts as natural sweeteners. Support or Funding Information This research was supported by the by the National Science Council of Taiwan (NSC Grant 106-2218-E-029-002-MY2). The screening of yeast transformants having its exg1Δ::kanMX4 replaced by DbExg1-His-URA3 cassette which has S I bioconversion activity A. Ura− and G418 resistance selection. B. The genotype confirmation of yeast transformants. Successful transformants have 3523 b.p product. Numbers represent candidates relative to candidates in A. C. Screening of transformants with siamenoside I production specificity. Peak 1, mogroside V; 2, siamenoside I; 3, mogroside IV; 4, mogroside IIIE. SEM images and size distribution of electrospun nanofibrous membrane: (Upper panel) PVA; (Lower panel) cross-lined PVA. SEM images and size distribution of electrospun nanofibrous membrane: (Upper panel) PVA with 0.5% of the enzyme; (Lower panel) cross-lined PVA with 0.5% of the enzyme. SEM images and size distribution of electrospun nanofibrous membrane: (Upper panel) PVA with 2.0 % of the enzyme; (Lower panel) cross-lined PVA with 2.0% of the enzyme. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Hydrolyzation of mogrosides: Immobilized β-glucosidase for mogrosides deglycosylation from Lo Han Kuo.

An immobilized enzyme system for bioconversion of Lo Han Kuo (LHK) mogrosides was established. β‐Glucosidase which was covalently immobilized onto the glass spheres exhibited a significant bioconversion efficiency from pNPG to pnitrophenol over other carriers. Optimum operational pH and temperature were determined to be pH 4 and 30°C. Results of storage stability test demonstrated that the glass sphere enzyme immobilization system was capable of sustaining more than 80% residual activity until 50 days, and operation reusability was confirmed for at least 10 cycles. The Michaelis constant (K m) of the system was determined to be 0.33 mM. The kinetic parameters, rate constant (K) at which Mogrosides conversion was determined, the τ 50 in which 50% of mogroside V deglycosylation/mogroside IIIE production was reached, and the τ complete of complete mogroside V deglycosylation/mogroside IIIE production, were 0.044/0.017 min−1, 15.6/41.1 min, and 60/120 min, respectively. Formation of the intermediates contributed to the kinetic differences between mogroside V deglycosylation and mogroside IIIE formation.

Mogroside IIIE attenuates gestational diabetes mellitus through activating of AMPK signaling pathway in mice

As one kind of complications of pregnancy, gestational diabetes mellitus (GDM) can influence the health of maternal-child in clinical practice. The C57 BL/KsJdb/+(db/+) mice, genetic GDM model, and C57 BL/KsJ+/+ (wild-type) mice were purchased and classified into three groups: normal pregnancy (C57 BL/KsJ+/+), GDM (C57 BL/KsJdb/+), and GDM plus Mogroside IIIE (20.0 mg/kg) group. GDM symptoms (maternal body weight, serum glucose, and insulin levels), glucose and insulin tolerance, and reproductive outcome (body weight at birth and litter size of offspring) were investigated. The inflammatory factors such as IL-1β, IL-6, and TNF-α in the serum and the pancreas were detected by ELISA and qRT-PCR, while the expression of pAMPK, AMPK, pHDAC4, HDAC4, and G6Pase in the livers were analyzed by Western Blot. Mogroside IIIE greatly improved glucose metabolism, insulin tolerance, and reproductive outcome of the GDM mice. Moreover, Mogroside IIIE treatment significantly decreased inflammatory factors expression and relieved GDM symptoms through enhanced AMPK activation, inhibited HDAC4 expression, and reduced production of G6Pase. The alleviation of GDM by Mogroside IIIE was mediated by elevated AMPK activation, which in turn inhibited HDAC4 phosphorylation, and eventually down-regulated G6Pase expression and activity.

Mogroside IIIE Attenuates LPS-Induced Acute Lung Injury in Mice Partly Through Regulation of the TLR4/MAPK/NF-κB Axis via AMPK Activation.

Acute lung injury (ALI) often leads to high mortality, and there is as yet no effective drug treatment. The present study aimed to investigate protective effects of mogroside IIIE (MGIIIE, a cucurbitane-type triterpenoid from Siraitia grosvenorii Fruits) in experimental ALI and its underlying mechanism. MGIIIE (1, 10 0r 20mg/kg) was orally administered for 1h before a single intratracheal administration of lipopolysaccharide (LPS, 5mg/kg). MGIIIE treatment dose-dependently suppressed pulmonary oedema, pro-inflammatory mediators (IL-1β, IL-6, TNF-α and HMGB1) release and higher MPO activity in lung tissues induced by LPS challenge. Molecular researches showed that mogroside IIIE (20mg/kg) not only increased the phosphorylation of adenosine 5'-monophosphate-activated protein kinase (AMPK) but suppressed the over-expression of toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (MyD88). In addition, MGIIIE also inhibited the activation of MAPKs and nuclear factor κB (NF-κB) signalling in lung tissues from LPS-challenged mice. Similar antiinflammatory effects of MGIIIE were obtained in LPS-treated macrophages. Compound C (a pharmacological AMPK inhibitor) obviously reversed the antiinflammatory effect of MGIIIE in LPS-induced ALI mice. Taken together, AMPK activation plays a crucial role in the antiinflammatory effects of MGIIIE in LPS-induced ALI by down-regulating TLR4/MAPK/NF-κB signalling pathways. Copyright © 2017 John Wiley & Sons, Ltd.

Mogroside IIIE, a Novel Anti-Fibrotic Compound, Reduces Pulmonary Fibrosis through Toll-Like Receptor 4 Pathways.

Idiopathic pulmonary fibrosis is a progressive fibrotic lung disease, and eventually most patients develop respiratory failure with a median survival rate of 2 to 3 years after diagnosis due to the lack of clinically effective therapies. Mogroside IIIE (MGIIIE), a cucurbitane-type compound, was isolated from Siraitia grosvenorii MGIIIE has shown the strongest inhibition of nitric oxide release, a crucial inflammatory factor, from lipopolysaccharide (LPS)-treated RAW264.7 cells compared with other mogrosides. In the pulmonary fibrosis mouse model induced by bleomycin, MGIIIE treatment attenuated pulmonary fibrosis, indicated as a reduction in myeloperoxidase activity, collagen deposition, and pathologic score. MGIIIE also significantly suppressed expression of several important fibrotic markers, e.g., α-smooth muscle actin, collagen I, transforming growth factor-β (TGF-β) signal, and metalloproteinases-9/tissue inhibitor of metalloproteinase-1. Furthermore, MGIIIE blocked tansdifferentiation of lung resident fibroblasts into myofibroblast-like cells induced by TGF-β or LPS and subsequently inhibited collagen production in lung fibroblasts. These data indicate that MGIIIE is a potent inhibitor for pulmonary fibrosis. In vitro and in vivo mechanistic studies have shown that MGIIIE significantly decreased expression of toll-like receptor 4 (TLR4) and its downstream signals of myeloid differentiation factor 88 (MyD88)/mitogen-activated protein kinase (MAPK), an inflammatory signal essential for extracellular matrix (ECM) deposition in pulmonary fibroblasts. Taken together, these results demonstrate that MGIIIE significantly prevents pulmonary fibrosis by inhibiting pulmonary inflammation and ECM deposition through regulating TLR4/MyD88-MAPK signaling. Our study suggests that MGIIIE may have therapeutic potential for treating pulmonary fibrosis in clinical settings.

Metabolites of Siamenoside I and Their Distributions in Rats.

Siamenoside I is the sweetest mogroside that has several kinds of bioactivities, and it is also a constituent of Siraitiae Fructus, a fruit and herb in China. Hitherto the metabolism of siamenoside I in human or animals remains unclear. To reveal its metabolic pathways, a high-performance liquid chromatography-electrospray ionization-ion trap-time of flight-multistage mass spectrometry (HPLC-ESI-IT-TOF-MSn) method was used to profile and identify its metabolites in rats. Altogether, 86 new metabolites were identified or tentatively identified, and 23 of them were also new metabolites of mogrosides. In rats, siamenoside I was found to undergo deglycosylation, hydroxylation, dehydrogenation, deoxygenation, isomerization, and glycosylation reactions. Among them, deoxygenation, pentahydroxylation, and didehydrogenation were novel metabolic reactions of mogrosides. The distributions of siamenoside I and its 86 metabolites in rat organs were firstly reported, and they were mainly distributed to intestine, stomach, kidney, and brain. The most widely distributed metabolite was mogroside IIIE. In addition, eight metabolites were bioactive according to literature. These findings would help to understand the metabolism and effective forms of siamenoside I and other mogrosides in vivo.

Minor cucurbitane-glycosides from fruits of Siraitia grosvenori (Cucurbitaceae).

From fruits of Siraitia grosvenori, a Chinese medicinal plant, a new minor glycoside and four known minor cucurbitane glycosides, siamenoside I (sweet), 11-oxo-mogroside V (sweet), and mogrosides IIE and IIIE (both tasteless) were isolated together with mogrosides IV and V (both sweet) previously isolated from this fruit by Arihara et al. Structure of the new testeless glycoside called mogroside III was elucidated as 3-O-β-D-glucopyranosyl- 24-O-β-gentiobiosyl-mogrol. The relative sweetness of siamenoside I to sucrose was estimated ×563, making this the sweetest compound among the cucurbitane-glycosides so far isolated. The structure-taste relationship of cucurbitane-glycosides is also described.