Red Mold Rice Research Articles

Mini-review: medication safety of red yeast rice products.

High lipid levels in the blood together with high blood pressure and diabetes are among the highest risks for coronary heart disease. In particular, elevated cholesterol levels promote the progression of atherosclerosis. Red yeast rice, also called red fermented rice or red mold rice, is used as a dietary supplement to lower cholesterol levels. It contains varying amounts of natural monacolin K, which is a structural homolog to lovastatin, and shows hypocholesterolemic properties comparable to synthetic statins. Despite being portrayed as a natural alternative, the potential of red yeast rice for side effects and interactions is comparable to statin drugs. Consumers need to be made aware of the varying monacolin K content in red yeast rice products and the insufficient long-term safety data regarding the potential risks of red yeast rice.

A New Azaphilone Derivative from the Monascus kaoliang Fermented Rice

Investigation of the 95% EtOH extract of red mold rice fermented with the red mutant of the fungus Monascus kaoliang led to the isolation of one new azaphilone derivative, namely monakaocinol (1). The structure was determined by analyses of mass spectrometry and 1D and 2D NMR data. The relative configuration of the isolated compound was assigned on the basis of 2D NOESY experiments.

Phytosomal bilayer-enveloped casein micelles for codelivery of monascus yellow pigments and resveratrol to breast cancer.

Multireservoir nanocarriers were fabricated for delivering antineoplastic drug cocktail from herbal and fungal origin. Monascus yellow pigments (MYPs), monascin and ankaflavin, were isolated from red-mold rice, and incorporated within casein micelles (CAS MCs) along with the herbal drug, resveratrol (RSV). Both drugs (MYPs and RSV) were simultaneously incorporated into the hydrophobic core of CAS MCs. Alternatively, MYPs-loaded CAS MCs were enveloped within RSV-phytosomal bilayer elaborating multireservoir nanocarriers. Cytotoxicity studies confirmed the superiority of multireservoir nanocarriers against MCF-7 breast cancer cells. The in vivo antitumor efficacy was revealed by reduction of the tumor volume and growth biomarkers. Multireservoir CAS nanocarriers for codelivery of both MYPs and RSV may be promising alternative to traditional breast cancer therapy.

Using millet as substrate for efficient production of monacolin K by solid-state fermentation of Monascus ruber

In this study, various grains such as rice, millet, corn, barley and wheat were used as raw materials for monacolin K production by solid-state fermentation of Monascus ruber. Among these substrates, millet was found to be the best one for monacolin K production, by which the yield reached 7.12mg/g. For enhanced monacolin K production, the effects of fermentation time, charge amount, initial moisture content and inoculum volume were systematically investigated in the solid-state fermentation of M. ruber. Moreover, complementary carbon source and nitrogen source were added for further improving the production of monacolin K. Results showed that the maximum production of monacolin K (19.81mg/g) could be obtained at the optimal conditions. Compared with the traditional red mold rice, using millet as substrate is promising for high production of monacolin K in the solid-state fermentation of M. ruber.

Structural Characterization of a New Sesquiterpene from the Fungal Strain Monascus kaoliang

A new sesquiterpene derivative, 10-methoxythujopsene (1), was isolated from the red mould rice of Monascus kaoliang. The structure was determined by extensive NMR and MS spectroscopic analysis. The relative configuration of 1 was established by the use of ROESY spectroscopic data.

Development of new red mold rice and determination of their properties

In order to analyze the effect of Chinese herbs on the three major enzyme activities: glucoamylase, protease and esterase as well as the biomass of the Monascus spp., a high quality Red mold rice (RMR) with Chinese herbs was developed. The new RMR demonstrated higher enzyme activity and biomass than the blank control. The glucoamylase, protease and esterase activity were increased to 518.03 U/g, 10.10 g/kg and 30.99 g/kg, respectively. Simultaneously, the biomass increased to 59.82 g/kg. With its application in rice wine, the alcohol yield ratio, total amino acid and the fragrance composition content increased obviously, respectively, and the organic acid content and the antioxidant capacity in vitro increased markedly.

New Metabolite Isolated from the Fungus Monascus pilosus

A new isochroman analogue, named monascuspilorin (1), was obtained from the n-BuOH-soluble fraction of the 95% EtOH extract of red mold rice fermented with the fungus Monascus pilosus BCRC 38072 (Eurotiaceae). The structure of the new compound was elucidated by comprehensive spectroscopic analysis, especially HR-ESI-MS and NMR experiments.

One New Compound from the Extract of the Fungus Monascus purpureus BCRC 31499

A chemical investigation on the n-BuOH-soluble fraction of the 95% EtOH extract of red mold rice fermented with the fungus Monascus purpureus BCRC 31499 (Eurotiaceae) has resulted in the isolation of one new isochroman derivative, designated as monascupurpurin (1). The structure was elucidated as rel-(2S)-4,6-dimethyl-2-hexyl-7-O-α-L-rhamnosylisochroman-5-ol (1) on the basis of extensive 1D and 2D NMR techniques, including COSY, HMBC, HMQC, and NOESY correlations, as well as HR-ESI-MS analysis and comparison of the spectroscopic data with those reported for structurally related compounds. This is the first report on Monascus metabolites with an isochroman glycoside.

Monascus Pigment Rubropunctatin: A Potential Dual Agent for Cancer Chemotherapy and Phototherapy.

The Monascus pigment, rubropunctatin, was extracted and purified from red mold rice (RMR), and its cytotoxic activities against human cervical carcinoma HeLa cells were studied under the conditions with or without light irradiation. The IC50 value of rubropunctatin against HeLa cells in the dark was 93.71 ± 1.96 μM (24 h), while the cytotoxic activity was enhanced more than 3 times (IC50 = 24.02 ± 2.17 μM) under light irradiation (halogen lamp, 500 W; wavelength, 597-622 nm; and fluence rate, 15 mW cm(-2), for 30 min). However, the IC50 value of rubropunctatin against the immortalized human cervical epithelial H8 cells was more than 300 μM, even under light irradiation, indicating that rubropunctatin has a favorable selectivity index (SI). Treatment of HeLa cells with rubropunctatin in the dark or under light irradiation resulted in a dose-dependent apoptosis, as validated by the increase in the percentage of cells in the sub-G1 phase and phosphatidylserine externalization, and the inductive effect on HeLa cell apoptosis was boosted by the light irradiation. In addition, treatment with rubropunctatin alone or under light irradiation was found to induce apoptosis in HeLa cells via the mitochondrial pathway, including loss of mitochondrial membrane potential, activation of caspase-3, caspase-8, and caspase-9, and increase of the level of intracellular reactive oxygen species (ROS). It was suggested that rubropunctatin could be a promising natural dual anticancer agent for photodynamic therapy and chemotherapy.

Neuroprotective effects of dimerumic acid and deferricoprogen from Monascus purpureus NTU 568-fermented rice against 6-hydroxydopamine-induced oxidative stress and apoptosis in differentiated pheochromocytoma PC-12 cells

abstractContext Oxidative stress plays a key role in neurodegenerative disorders, including Parkinson’s disease (PD). Rice fermented with Monascus purpureus Went (Monascaceae) NTU 568 (red mould rice) was found to contain antioxidants, including dimerumic acid (DMA) and deferricoprogen (DFC).Objective The effects of DMA and DFC on 6-hydroxydopamine (6-OHDA)-induced cytotoxicity and potential protective mechanisms in differentiated PC-12 pheochromocytoma cells were investigated.Materials and methods DMA (0–60 μM) or DFC (0–10 μM) was co-treated with 6-OHDA (200 μM, 24 h exposure) in differentiated PC-12 cells. Cell viability and intercellular reactive oxygen species (ROS) were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and 2′,7′-dichlorofluorescein-diacetate (DCFH-DA) assays, respectively. Cell apoptosis was determined by DNA fragmentation analysis and propidium iodide staining by flow cytometry. Western blot analysis was used to measure the levels of cell protein expression.Results DMA and DFC significantly increased cell viability to 72% and 81% in 6-OHDA-induced differentiated PC-12 cell cultures, respectively. Furthermore, DMA and DFC reduced 6-OHDA-induced formation of extracellular and intercellular ROS by 25% and 20%, respectively, and decreased NADPH oxidase-2 expression in differentiated PC-12 cells. DMA and DFC inhibited 6-OHDA-induced apoptosis and decreased activation of caspase-3 via regulation of Bcl-2-associated X protein (Bax) and Bcl-2 protein expression in differentiated PC-12 cells.Conclusion DMA and DFC may protect against 6-OHDA toxicity by inhibiting ROS formation and apoptosis. These results showed that the metabolites from M. purpureus NTU 568 fermentation were potential therapeutic agents for PD induced by oxidative damage and should be encouraged for further research.

Effects of Red Mold Rice Produced from Monascus purpureus CMU002U on Growth Performances and Antioxidant Activity of Japanese Quail

Effects of Red Mold Rice Produced from Monascus purpureus CMU002U on Growth Performances and Antioxidant Activity of Japanese Quail

A soil bacterium Rhizobium borbori and its potential for citrinin-degrading application

Red mold rice is the fermentation product of ordinary white rice with various Monascus spp. and is popular in Asian countries as a food additive and for its beneficial effects on human health. However, most Monascus spp. not only produce valuable secondary metabolites, such as monacolin K, but also the mycotoxin citrinin, which limits the applicability of red mold rice in medical therapy and food production. The aim of this study was to isolate, screen and identify citrinin-degrading bacteria that can be used for citrinin degradation in industrial and medical applications. A total of 96 bacteria were obtained from 74 samples of soil, water and wastewater collected from Chiang Mai, northern Thailand. Of these, 14 isolates exhibited growth in a mineral medium containing 5 ppm crude citrinin, among which only five demonstrated citrinin-degrading ability without any loss of monacolin K after culture at 30 °C for 72 h. Isolates PS21 and PS45 degraded citrinin the fastest of all five isolates and also retained the lowest residual citrinin concentration. Based on 16S rRNA gene sequence analysis, isolates PS21 and PS45 were identified as Enterococcus cloacae and Rhizobium borbori, respectively. When R. borbori PS45 and E. cloacae PS21 were cultured in a mineral medium containing 5 ppm pure citrinin at 30 °C for 120 h, the residual citrinin contents were 1.83 and 2.82 ppm, respectively. The citrinin-degrading activities of both isolates were found predominantly in the cell-free supernatant. Based on its higher citrinin-degrading activity and safety, we propose R. borbori PS45 as a suitable microorganism for citrinin reduction in red mold rice products.

Edible Filamentous Fungi from the Species Monascus: Early Traditional Fermentations, Modern Molecular Biology, and Future Genomics

AbstractMonascus spp. are filamentous fungi famous for their fermented products, especially red mold rice (RMR), a traditional fermented food in East Asian areas with a very long edible history documented back to the Han dynasty (BC 202‐AD 220) in China. Nowadays, RMR and its related products involve a very large industry from artisanal traditional fermentations to food companies to medicine manufacturers, which are distributed worldwide. Modern studies have shown that Monascus spp. are able to produce abundant beneficial secondary metabolites, such as monacolins (cholesterol‐lowering agents), γ‐amino butyric acid (an antihypertensive substance), dimerumic acid (an antioxidant), and pigments (food‐grade colorants), and some strains can also secrete citrinin, a nephrotoxic metabolite. Monascus‐related studies have received much attention because of their wide applications. However, to our knowledge, no systematic review on the progress of Monascus research has ever been published. In this review, the progress of research on Monascus is summarized into 3 stages: Monascus fermentation, Monascus molecular biology, and Monascus genomics. This review covers the past history, current status, and future direction of Monascus research, contributing to a comprehensive understanding of Monascus research progress.

Proteomic Analysis of Human Breast Cancer Cells Treated with <i>Monascus</i>-Fermented Red Mold Rice Extracts

Monascus-fermented red mold rice (RMR) extracts have recently been proved to possess anticancer activities, but the anticancer effect of breast cancer cells treated with RMR is not yet well understood. In an effort to identify proteins that may be involved in RMR-induced breast cancer cell death, we employed proteomic analysis coupled with LC-nanoESI-MS/MS on MCF-7 cells exposed to two RMR extracts, ethyl acetate extract (EAE) and ethanol extract (EE). Sixupregulated proteins were identified in EAE-induced MCF-7 cells, including glyceraldehyde-3-phosphate dehydrogenase, pyruvate dehydrogenase E1 component, heat shock 27 kDa protein 1, cathepsin D, protein disulphideisomerase A3, and prohibitin. The upregulated expression of other 4 proteins was seen in EE-induced MCF-7 cells, including alloalbuminvenezia, annexin A5, endoplasmic reticulum protein 29 isoform 1 precursor, and cellular retinoic acid binding protein 2. The same two down-regulated proteins were identified on cells after treatment with both extracts, dermcidin preproprotein and poly(rC) binding protein 1. These proteins have been implicated in proapoptotic regulation, stress modulation, tumor suppression, and survival factor activity. These analysescould provide valuable information for further exploration of RMR as a promising chemopreventive agent against breast cancer.

Neurocytoprotective Effects of Aliphatic Hydroxamates from Lovastatin, a Secondary Metabolite from Monascus-Fermented Red Mold Rice, in 6-Hydroxydopamine (6-OHDA)-Treated Nerve Growth Factor (NGF)-Differentiated PC12 Cells.

Lovastatin, a secondary metabolite isolated from Monascus-fermented red rice mold, has neuroprotective activity and permeates the blood-brain barrier. The aim of this study was to enhance the activity of lovastatin for potential use as a treatment for neuronal degeneration in Parkinson's disease. Six lovastatin-derived compounds were semisynthesized and screened for neurocytoprotective activity against 6-hydroxydopamine (6-OHDA)-induced toxicity in human neuroblastoma PC12 cells. Four compounds, designated as 3a, 3d, 3e, and 3f, significantly enhanced cell viability. In particular, compound 3f showed excellent neurocytoprotective activity (97.0 ± 2.7%). Annexin V-FITC and propidium iodide double staining and 4',6-diamidino-2-phenylindole staining indicated that compound 3f reduced 6-OHDA-induced apoptosis in PC12 cells. Compound 3f also reduced caspase-3, -8, and -9 activities, and intracellular calcium concentrations elevated by 6-OHDA in a concentration-dependent manner, without inhibiting reactive oxygen species generation. JC-1 staining indicated that compound 3f also stabilized mitochondrial membrane potential. Thus, compound 3f may be used as a neurocytoprotective agent. Future studies should investigate its potential application as a treatment for Parkinson's disease.

Effect of white wheat bread and white wheat bread added with bioactive compounds on hypercholesterolemic and steatotic mice fed a high-fat diet.

The effects of white wheat bread and white wheat bread added with a bioactive compound mixture (Cyclanthera pedata, Glycine max, Monascus-fermented red mold rice, Cynara scolymus and Medicago sativa) were examined on hypercholesterolemic and steatotic mice, divided into four groups: control diet (CTR), high-fat diet (HFD), high-fat diet with white wheat bread added with 1.5 g kg(-1) of mixture (HFD+AB) and high-fat diet with white wheat bread (HFD+B). Total serum cholesterol in the HFD+AB and HFD+B groups and hepatic triglycerides in the HFD+AB group decreased compared with the HFD group. Liver histology confirmed lower lipid drop accumulation in the HFD+AB group than in the HFD and HFD+B groups. HFD+AB caused a 7.0-fold increase and a 3.5-fold reduction in CYP7A1 and SREBP-1c gene expression respectively compared with the HFD group. Moreover, the HFD+B group showed a 2.2-, 8.4- and 1.5-fold increase in HMG CoA reductase, CYP7A1 and LDLr gene expression respectively compared with the HFD group. Both the white wheat bread and the added white wheat bread induced cholesterol reduction by increasing CYP7A1. Moreover, the added white wheat bread improved steatosis by decreasing SREBP-1c gene expression.

Safety and mutagenicity evaluation of red mold dioscorea fermented from Monascus purpureus NTU 568

Monascus-fermented products, including red mold rice and red mold dioscorea, have been developed as functional foods with many health benefits. We performed safety and mutagenic evaluations on red mold dioscorea powder (RMDP) fermented from Monascus purpureus NTU 568. The results of Ames test using Salmonella typhimurium strains TA97a, TA98, TA100, TA102, and TA1535 showed that RMDP (⩽5mg/plate) was not mutagenic. The mammalian chromosomal aberration test showed that the number of Chinese hamster ovary cells with abnormal chromosomes was <3% after RMDP treatment (maximum concentration: 5mg/mL). Imprinting control region mice were used to estimate the genotoxicity of RMDP. Compared with the control, high-dose RMDP administration (2000mg/kg) did not show significant differences in the number of reticulocytes or the occurrence of micronucleated reticulocytes. A 28-day oral toxicity assay in Sprague–Dawley rats was performed to investigate the no observed adverse effect level of RMDP. Compared with the control, high-dose RMDP administration (2000mg/kg) caused no toxicological responses such as mortality, variation in body weight, or toxicopathologic lesions. Thus, RMDP from M. purpureus NTU 568 shows no significant mutagenic or toxic effects.

Monascus ruber의 액체배양을 통한 색소 생산 및 고체발효를 통한 홍국쌀의 monacolin K 생산 특성

The growth characteristics and production of color pigments by Monascus strains were investigated during liquid culture, and production of monacolin K in red mold rice was carried out by solid state fermentation. Four different Monascus ruber strains were cultured in potato dextrose yeast extract broth (PDYB) media at <TEX>$25^{\circ}C$</TEX> for 15 days. The high producing strain for red pigment was not corresponded to the strain for yellow pigment. Production of red pigment was high in the strain causing the fast pH change in culture broth. Production of monacolin K in red mold rice by solid state fermentation was influenced by a combination of wet cell weight and spore density in inoculum by liquid culture. Most strains showed the high production of monacolin K in red mold rice, when submerged fermentation was carried out for 5 days as inoculum for solid state fermentation. These results suggest that submerged fermentation period of inoculum have an effect on the production of monacolin K in red mold rice by solid state fermentation, and monacolin K in red mold rice could be increased by controlling the condition of submerged fermentation for inoculum.

Effect of red mold rice on blood coagulation and anticoagulation factors in a rat model of hyperlipidemia

The bioactive monacolin K found in red mold rice (RMR) is identical in structure to lovastatin, and has the effect of lowering the cholesterol level in human blood. Statins not only lower atherosclerosis caused by thrombus, but also reduce the death rate from coronary artery disease. This study was designed to evaluate the effects of Monascus purpureus NTU 803 fermented-RMR on blood coagulation and anticoagulation factors in hyperlipidemic rats. Rats were fed a high-lipid diet for 8weeks to induce hyperlipidemia and fed various doses of RMR. The results showed that RMR prolonged the bleeding time of the rats, inhibited platelet coagulation, lowered the fibrinogen content in the blood, increased the antithrombin III and protein C content in the blood, and reduced lipid plaque in the aorta. RMR was effective for preventing the formation of thrombus and reducing the occurrence of atherosclerosis.

Development of a Rapid LC/DAD/FLD/MSn Method for the Simultaneous Determination of Monacolins and Citrinin in Red Fermented Rice Products

Red fermented rice is used worldwide by many patients as an alternative therapy for hyperlipidemia; however, the discovery of a toxic fermentation byproduct, citrinin, causes much controversy about the safety of red mold rice products. A new and fast high-performance liquid chromatography method was developed and validated for simultaneous determination of cholesterol-lowering compounds monacolin K (lovastatin), monacolin K hydroxy acid, and other monacolins present in red fermented rice as well as nephrotoxic mycotoxin citrinin in a single run using connected diode array and fluorescence and mass spectrometric detectors. The proposed method was successfully applied for the analysis of red fermented rice food samples and various dietary supplements also containing other natural lipid-lowering agents. The deviations between label content and levels of active compounds found in investigated samples as well as high batch-to-batch variation found in one product indicate that the regular quality control of red fermented rice products is of great importance.