Screening Antibacterial Photodynamic Effect of Monascus Red Yeast Rice (Hong-Qu) and Mycelium Extracts.

The fungus Monascus is a well-known source of secondary metabolites with interesting pharmaceutical and nutraceutical applications. In particular, Monascus pigments possess a wide range of biological activities (e.g. antimicrobial, antioxidant, anti-inflammatory or antitumoral). To broaden the scope of their possible application, this study focused on testing Monascus pigment extracts as potential photosensitizing agents efficient in antimicrobial photodynamic therapy (aPDT) against bacteria. For this purpose, eight different extracts of secondary metabolites from the liquid- and solid-state fermentation of Monascus purpureus DBM 4360 and Monascus sp. DBM 4361 were tested against Gram-positive and Gram-negative model bacteria, Bacillus subtilis and Escherichia coli and further screened for ESKAPE pathogens, Staphylococcus aureus and Pseudomonas aeruginosa. To the bacterial culture, increasing concentration of extracts was added and it was found that all extracts showed varying antimicrobial activity against Gram-positive bacteria in dark, which was further increased after irradiation. Gram-negative bacteria were tolerant to the extracts' exposure in the dark but sensitivity to almost all extracts that occurred after irradiation. The Monascus sp. DBM 4361 extracts seemed to be the best potential candidate for aPDT against Gram-positive bacteria, being efficient at low doses, i.e. the lowest total concentration of Monascus pigments exhibiting aPDT effect was 3.92 ± 1.36 mg/L for E. coli. Our results indicate that Monascus spp., forming monascuspiloin as the major yellow pigment and not-forming mycotoxin citrinin, is a promising source of antimicrobials and photoantimicrobials.

Structural elucidation and complete NMR spectral assignments of Monascus monacolin analogues.

Three new monacolin analogues, 3,6-dihydroxy-monacolin P (1), 6-methoxy monacolin S (2), and 6-methoxy dehydromonacolin S (3), were isolated from a fraction that strongly inhibited 3-hydroxy-3-methylglutaryl-CoA reductase from the ethyl acetate portion of red yeast rice ethanol extract. Their structures were determined through a combination of 1D and 2D NMR experiments, mass spectrometry analysis, and known literature reports.

Unveiling the potential of red koji polysaccharides: biosynthesis, extraction, and multifaceted biological activities.

Red koji polysaccharides, derived from the fermentation of Monascus, have been recognized for their health-enhancing properties. This article reviews their structural characteristics, biosynthesis pathways, and biological activities. It emphasizes the need for sustainable practices in fermentation and the optimization of extraction methods for scalable production. The significance of exploring the molecular mechanisms involved in their biosynthesis is also highlighted to enhance yield and efficiency. Research indicates that red koji polysaccharides possess diverse biological functions, beneficial for pharmaceutical applications due to their health benefits and minimal toxicity. The review points out the necessity for more detailed studies on key enzymes and genes in biosynthesis to improve production methods. It also identifies the current challenges in production scalability and extraction efficiency. Furthermore, while these polysaccharides show potential in pharmaceuticals, their clinical efficacy and mechanism of action in human subjects require further investigation. The review briefly explores potential structural modifications to improve their biological activities. The review concludes that red koji polysaccharides hold significant untapped potential, particularly in drug formulation. Future research should focus on overcoming current production and application challenges, including conducting clinical trials to validate their efficacy and exploring structural modifications for enhanced therapeutic benefits. This comprehensive understanding of red koji polysaccharides paves the way for their expanded application in the pharmaceutical industry. © 2024 Society of Chemical Industry.

Toward improvements for enhancement the productivity and color value of Monascus pigments: a critical review with recent updates.

Monascus pigments are a kind of high-quality natural edible pigments fermented by Monascus filamentous fungi, which have been widely used in food, cosmetics, medicine, textiles, dyes and chemical industries as active functional ingredients. Moreover, Monascus pigments have a good application prospect because of a variety of biological functions such as antibacterial, antioxidation, anti-inflammatory, regulating cholesterol, and anti-cancer. However, the low productivity and color value of pigments restrict their development and application. In this review, we introduced the categories, structures, biosynthesis and functions of Monascus pigments, and summarized the current methods for improving the productivity and color value of pigments, including screening and mutagenesis of strains, optimization of fermentation conditions, immobilized fermentation, mixed fermentation, additives, gene knockout and overexpression technologies, which will help to develop the foundation for the industrial production of Monascus pigments.

Core Steps to the Azaphilone Family of Fungal Natural Products.

Azaphilones are a family of polyketide-based fungal natural products that exhibit interesting and useful bioactivities. This minireview explores the literature on various characterised azaphilone biosynthetic pathways, which allows for a proposed consensus scheme for the production of the core azaphilone structure, as well as identifying early diversification steps during azaphilone biosynthesis. A consensus understanding of the core enzymatic steps towards a particular family of fungal natural products can aid in genome-mining experiments. Genome mining for novel fungal natural products is a powerful technique for both exploring chemical space and providing new insights into fungal natural product pathways.

Monalbidins A-E, Decalins with Potential Cytotoxic Activities from Marine Derived Fungus Monascus albidus BB3.

Five new decalins, monalbidins A-E (1, 2 and 7-9), together with 16 known compounds (3-6 and 10-21), were isolated from the AcOEt extract of marine derived fungus Monascus albidus BB3 cultured in GPY medium. Among the known compounds, 1-hydroxymonacolin L (11), dehydromonacolin J (15), 8-O-acetylmonacolin J (19) and O-acetylmonacolin K (21) were separated from natural sources for the first time. Their structures were determined by comprehensive analysis on the 1D and 2D NMR, HR-ESI-MS, UV and IR data, and their absolute configurations were assigned by experimental and calculated ECD data, and X-ray single-crystal diffraction analysis. Monalbidins C and D (7 and 8), monacolin K methyl ester (13), dehydromonacolin L (14), dehydromonacolin K (16), monacolin K (20) and O-acetylmonacolin K (21) showed moderate cytotoxicity against human cancer cell lines SUNE1, HepG2, QGY7701, HCT116 and MDA-MB-231.

Effects of Various Rice-Based Raw Materials on Enhancement of Volatile Aromatic Compounds in Monascus Vinegar.

Monascus vinegar (MV), during whose brewing process Monascus spp. and polished rice (PR) are normally used as the starter and the raw material, respectively, is one of the traditional vinegars in China. In this study, the effects of three raw materials, including PR, unhusked rice (UR), and germinated UR (GR), on MV volatile compounds have been investigated. The results revealed that MV of GR (GMV), and its intermediate Monascus wine (GMW), exhibited the highest amount of aroma, not only in the concentrations but also in the varieties of the aromatic compounds mainly contributing to the final fragrance. Especially after three years of aging, the contents of benzaldehyde and furfural in GMV could reach to 13.93% and 0.57%, respectively, both of which can coordinate synergistically on enhancing the aroma. We also found that the filtering efficiency was significantly improved when UR and GR were applied as the raw materials, respectively. Therefore, GR might be more suitable raw materials for MV.

Monascin and Ankaflavin of Monascus purpureus Prevent Alcoholic Liver Disease through Regulating AMPK-Mediated Lipid Metabolism and Enhancing Both Anti-Inflammatory and Anti-Oxidative Systems.

Alcohol metabolism causes an excessive accumulation of liver lipids and inflammation, resulting in liver damage. The yellow pigments monascin (MS) and ankaflavin (AK) of Monascus purpureus-fermented rice were proven to regulate ethanol-induced damage in HepG2 cells, but the complete anti-inflammatory and anti-fatty liver mechanisms in the animal model are still unclear. This study explored the roles of MS and AK in improving alcoholic liver injury. MS and AK were simultaneously fed to evaluate their effects and mechanisms in C57BL/6J mice fed the Lieber-DeCarli liquid alcohol diet for 6 weeks. The results indicated that MS and AK significantly reduced the serum aspartate aminotransferase and alanine aminotransferase activity, as well as the total liver cholesterol and triglyceride levels. The histopathological results indicated that MS and AK prevented lipid accumulation in the liver. MS and AK effectively enhanced the activity of antioxidant enzymes and reduced the degree of lipid peroxidation; AK was particularly effective and exhibited a superior preventive effect against alcoholic liver injury and fatty liver. In addition to inhibiting the phosphorylation of the MAPK family, MS and AK directly reduced TNF-α, IL-6, and IL-1ß levels, thereby reducing NF-κB and its downstream iNOS and COX-2 expressions, as well as increasing PPAR-γ, Nrf-2, and HO-1 expressions to prevent liver damage. MS and AK also directly reduced TNF-α, IL-6, and IL-1ß expression, thereby reducing the production of NF-κB and its downstream iNOS and COX-2, and increasing PPAR-γ, Nrf-2, and HO-1 expressions, preventing alcohol damage to the liver.

Ultrasonic and enzymatic pretreatments of Monascus fermentation byproduct for a sustainable production of Bacillus subtilis.

BACKGROUND: Monascus fermentation byproduct (MFB) is a biowaste generated after food colorants are extracted. Using MFB to produce probiotics (Bacillus subtilis) is a sustainable way for the entire production to be used as food or animal feed additives. However, due to the rigidity of the Monascus mycelium cell wall, B. subtilis cannot sufficiently utilize the nutrients in MFB, leading to low biomass production efficiency. We studied the effects of ultrasonic treatment, papain, ß-glucanase, and chitosanase, and their combinations on improving the levels of soluble components from MFB. The effects of these treatments on mycelium cell walls were visualized using scanning electron microscopy, and their influence on B. subtilis production was analyzed. RESULTS: Ultrasonic treatment increased the soluble components by 210 g kg-1 , including 50 g kg-1 protein and 120 g kg-1 carbohydrates. An enzyme mixture increased the soluble components by 160 g kg-1 , including 30 g kg-1 protein and 90 g kg-1 carbohydrates. The combination of the two methods achieved the highest increase of soluble components (up to 400 g kg-1 ) leading to a maximum B. subtilis production of 1 × 1011 colony-forming unit mL-1 . This yield was about 20 times greater than that using untreated MFB and about eight times greater than treatments using only ultrasonic or enzymatic methods. CONCLUSION: The productivity of B. subtilis production using MFB as the sole medium can be greatly improved by ultrasound or enzymes, which cause the release of intercellular components or cell wall components. © 2020 Society of Chemical Industry.

Solid-state fermentation for enhanced production of selenium nanoparticles by gamma-irradiated Monascus purpureus and their biological evaluation and photocatalytic activities.

Selenium nanoparticles (SeNPs) were successfully synthesized using the culture extract of Monascus purpureus ATCC16436 grown on sugarcane bagasse under solid-state fermentation. The rapid synthesis of SeNPs was completed after 30 min as confirmed by UV-Vis spectroscopy. Functional groups present in the synthesized SeNPs samples were confirmed by Fourier transform infrared spectroscopy. The synthesized SeNPs showed a single-phase crystalline structure. Transmission electron microscope revealed the spherical shape and the mean particle size was 46.58 nm. Dynamic light scattering analysis showed that the synthesized SeNPs were monodispersed and the recorded polydispersity index value was 0.205. Zeta potential value of - 24.01 mV indicated the high stability of SeNPs. Besides, the biological activities of antioxidant, anticancer and antimicrobial as well as the photocatalytic activities were also studied. SeNPs showed promising antioxidant activity with 50% inhibitory concentration of 85.92 µg mL-1. Based on the MTT assay, SeNPs inhibited the proliferation of normal human melanocytes, human breast and liver cancer cell lines with 50% inhibitory concentrations of 45.21, 61.86 and 200.15 µg mL-1, respectively. SeNPs showed broad spectrum of antimicrobial potential against the tested human and plant pathogens. SeNPs showed efficient degradation of methylene blue dye. Moreover, the effect of gamma irradiation on the production enhancement of SeNPs was also adopted. Exposure of the fungal spores to gamma rays at 1000 Gy increased the yield of SeNPs to approximately fivefold. Hence, this study suggests a new and alternate approach with the excellent biotechnological potentiality for the production of SeNPs.

Effect of nonionic surfactant Brij 35 on morphology, cloud point, and pigment stability in Monascus extractive fermentation.

BACKGROUND: Nonionic surfactant Brij 35 in submerged fermentation of Monascus can significantly increase Monascus pigment yield. Here, the effects of nonionic surfactant Brij 35 on Monascus pigment secretion in extractive fermentation are discussed in terms of cell morphology, cloud point change, and pigment stability. RESULTS: At Brij 35 concentrations up to 32 g L-1 , the higher concentrations led to the loosening of the network structure on the surface of the fungal wall, enhanced cell wall permeability, and increased abundance of lipid droplets. Alternatively, when the concentration of Brij 35 exceeded 32 g L-1 , a large amount of substances accumulated on the surface of the fungal wall, permeability reduced, and the degree of oil droplet dispersion in cells decreased. Further, during extractive fermentation, Brij 35 induced formation of a grid structure on the fungal wall surface beginning on day 2, increased the number of intracellular lipid droplets, and promoted intracellular pigment secretion into the extracellular environment. When the cloud point temperature in the fermentation system approached that of fermentation, the nonionic surfactant exhibited stronger Monascus pigment extraction capacity, thereby enhancing pigment yield. Hence, Brij 35 can improve pigment stability and effectively reduce damage caused by natural factors, such as light and temperature. CONCLUSION: Brij 35 promotes the secretion of pigment by changing the fungal wall structure and cloud point, as well as by improving pigment stability. © 2020 Society of Chemical Industry.

Nature and nurture: confluence of pathway determinism with metabolic and chemical serendipity diversifies Monascus azaphilone pigments.

Covering: up to June 2018 Understanding the biosynthetic mechanisms that generate the astounding structural complexity and variety of fungal secondary metabolites (FSMs) remains a challenge. As an example, the biogenesis of the Monascus azaphilone pigments (MonAzPs) has remained obscure until recently despite the significant medical potential of these metabolites and their long history of widespread use as food colorants. However, a considerable progress has been made in recent years towards the elucidation of MonAzPs biosynthesis in various fungi. In this highlight, we correlate a unified biosynthetic pathway with the diverse structures of the 111 MonAzPs congeners reported until June 2018. We also discuss the origins of structural diversity amongst MonAzPs analogues and summarize new research directions towards exploring novel MonAzPs. The case of MonAzPs illuminates the various ways that FSMs metabolic complexity emerges by the interplay of biosynthetic pathway determinism with metabolic and chemical serendipity.

Effects of nonionic surfactants on pigment excretion and cell morphology in extractive fermentation of Monascus sp. NJ1.

BACKGROUND: Different nonionic surfactants in submerged fermentation of Monascus sp. demonstrate significant differences regarding increasing pigment yield. In this study, 15 surfactants from five series were analyzed to investigate the influence of nonionic surfactants on Monascus pigments, with the aim of simultaneously obtaining a novel nonionic surfactant. RESULTS: Addition of the novel surfactant Brij 35 greatly enhanced pigment excretion and demonstrated good biocompatibility. Extracellular red, orange and yellow pigments increased by 1.47-, 1.71- and 2.07-fold respectively. Production of extracellular pigments was not only related to the hydrophile-lipophile balance value (HLB) but also affected by the cloud point temperature (CP) of the fermentation medium. It was found that nonionic surfactants can improve cell membrane permeability and cell storage capacity by modifying the cell walls of Monascus mycelium and by increasing lipid droplet levels, enhancing pigment excretion. Different nonionic surfactants modify Monascus mycelium to different degrees. CONCLUSION: The novel surfactant Brij 35, which has good capacity for pigment extraction and biocompatibility, was identified in the analysis. The effects of nonionic surfactants on the secretion of pigments are related to not only the modification of the cell wall and internal structure but also the CP and HLB. © 2018 Society of Chemical Industry.

Transcriptional regulation contributes more to Monascus pigments diversity in different strains than to DNA sequence variation.

Monascus azaphilone pigments, including red, orange, and yellow, are world-famous food colorants. However, the pigments produced by different Monascus species vary in yields and compositions. The underlying mechanism is unclear. In this study, four wild-type Monascus strains, namely M. anka M7, M. purpureus M9, M. ruber C100, and M. aurantiacus M15, were selected as research objects according to the diversification of their pigments fermented in the same mediums and conditions. Twenty-three 3 kbp segments (300 bp overlap with adjacent segments) of the pigment gene cluster were amplified, sequenced, and assembled into the DNA sequences of the clusters. The DNA sequences of pigment biosynthetic gene clusters of the four strains showed 99.94% similarity according to the results of multiple alignment. The expression levels of 17 pigment biosynthetic genes of four strains were determined by using real-time quantitative PCR. The transcriptional regulation contributed more than the DNA sequence variation in Monascus pigments metabolism. Our result gives insight into the study of Monascus pigment biosynthesis.

Production of Monascus pigments as extracellular crystals by cell suspension culture.

It is generally accepted that Monascus pigments are predominantly cell-bound, including both intracellular and surface-bound pigments. This long-term misconception was corrected in the present work. Production of extracellular crystal pigments by submerged culture of Monascus sp. was confirmed by microscopic observation and collection of Monascus pigments from extracellular broth by direct membrane filtration. Following up the new fact, the bioactivity of mycelia as whole-cell biocatalyst for biosynthesis and biodegradation of Monascus pigments had been detailedly examined in both an aqueous solution and a nonionic surfactant micelle aqueous solution. Based on those experimental results, cell suspension culture in an aqueous medium was developed as a novel strategy for accumulation of high concentration of Monascus pigments. Thus, glucose feeding during submerged culture in the aqueous medium was carried out successfully and high orange Monascus pigments concentration of near 4 g/L was achieved.

Extracellular red Monascus pigment-mediated rapid one-step synthesis of silver nanoparticles and its application in biomedical and environment.

The development of a safe and eco-friendly method for metal nanoparticle synthesis has an increasing demand, due to emerging environmental and biological harms of hazardous chemicals used in existing nanosynthesis methods. The present investigation reports a rapid one-step, eco-friendly and green approach for the formation of nanosized silver particles (AgNPs) using extracellular non-toxic-colored fungal metabolites (Monascus pigments-MPs). The formation of nanosized silver particles utilizing Monascus pigments was confirmed after exposure of reaction mixture to sunlight, by visually color change and further established by spectrophotometric analysis. The size, shape, and topography of synthesized MPs-AgNPs were well-defined using different microscopic and spectroscopic techniques, i.e., FE-SEM, HR-TEM, and DLS. The average size of MPs-AgNPs was found to be 10-40 nm with a spherical shape which was highly stable and dispersed in the solution. HR-TEM and XRD confirmed crystalline nature of MPs-AgNPs. The biocidal potential of MPs-AgNPs was evaluated against three bacterial pathogens such as Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus and it was observed that the MPs-AgNPs significantly inhibited the growth of all three bacterial pathogens. The anti-biofilm activity of MPs-AgNPs was recorded against antibiotic-resistant P. aeruginosa. Besides, the colorimetric metal sensing using MPs-AgNPs was studied. Among the metals tested, the selective Hg2+-sensing potential at micromolar concentration was observed. In conclusion, this is the rapid one-step (within 12-15 min), environment-friendly method for synthesis of AgNPs and synthesized MPs-AgNPs could be used as a potential antibacterial agent against antibiotic-resistant bacterial pathogens.

Four new monacolin analogs from Monascus purpureus-fermented rice.

Four new monacolin analogs, monacolin T (1), monacolin U (2) 6a-O-methyl-4,6-dihydromonacolin L (3), and 6a-O-ethyl-4,6-dihydromonacolin L (4) were isolated from the ethanolic extract of Monascus purpureus-fermented rice. Their structures were determined by a combination of 1D, 2D NMR experiments (1H-1HCOSY, HSQC, HMBC, and ROESY), and mass spectrometry. In vitro cytotoxic assay, all compounds were inactive at the concentration of 10 µM.

Evaluating Antitumor and Antioxidant Activities of Yellow Monascus Pigments from Monascus ruber Fermentation.

Yellow Monascus pigments can be of two kinds: Natural and reduced, in which natural yellow Monascus pigments (NYMPs) attract widespread attention for their bioactivities. In this study, the antioxidative and antibreast cancer effects of the water-soluble NYMPs fermented by Monascus ruber CGMCC 10910 were evaluated. Results showed that water-soluble NYMPs had a significantly improved antioxidative activities compared to the reduced yellow Monascus pigments (RYMPs) that were chemically derived from orange or red Monascus pigments. Furthermore, NYMPs exhibited a concentration-dependent inhibition activity on MCF-7 cell growth (p < 0.001). After a 48-h incubation, a 26.52% inhibition yield was determined with 32 µg/mL of NYMPs. NYMPs also significantly inhibited the migration and invasion of MCF-7 cells. Mechanisms of the activities were associated with a down-regulation of the expression of matrix metalloproteinases and vascular endothelial growth factor. Rather than being alternatively used as natural colorants or antioxidants, this work suggested that NYMPs could be selected as potential functional additives in further test of breast cancer prevention and adjuvant therapy.

Changing oxidoreduction potential to improve water-soluble yellow pigment production with Monascus ruber CGMCC 10910.

BACKGROUND: Monascus pigments are widely used in the food and pharmaceutical industries due to their safety to human health. Our previous study found that glucose concentration induced extracellular oxidoreduction potential (ORP) changes could influence extracellular water-soluble yellow pigment production by Monascus ruber CGMCC 10910 in submerged fermentation. In this study, H2O2 and dithiothreitol (DTT) were used to change the oxidoreduction potential for investigating the effects of oxidative or reductive substances on Monascus yellow pigment production by Monascus ruber CGMCC 10910. RESULTS: The extracellular ORP could be controlled by H2O2 and DTT. Both cell growth and extracellular water-soluble yellow pigment production were enhanced under H2O2-induced oxidative (HIO) conditions and were inhibited under dithiothreitol-induced reductive conditions. By optimizing the amount of H2O2 added and the timing of the addition, the yield of extracellular water-soluble yellow pigments significantly increased and reached a maximum of 209 AU, when 10 mM H2O2 was added on the 3rd day of fermentation with M. ruber CGMCC 10910. Under HIO conditions, the ratio of NADH/NAD+ was much lower than that in the control group, and the expression levels of relative pigment biosynthesis genes were up-regulated; moreover, the activity of glucose-6-phosphate dehydrogenase (G6PDH) was increased while 6-phosphofructokinase (PFK) activity was inhibited. CONCLUSIONS: Oxidative conditions induced by H2O2 increased water-soluble yellow pigment accumulation via up-regulation of the expression levels of relative genes and by increasing the precursors of pigment biosynthesis through redirection of metabolic flux. In contrast, reductive conditions induced by dithiothreitol inhibited yellow pigment accumulation. This experiment provides a potential strategy for improving the production of Monascus yellow pigments.

Monascustin, an Unusual γ-Lactam from Red Yeast Rice.

Monascustin (1), an unusual γ-lactam, was isolated from an ethanol extract of the Monascus purpureus fermented rice. Its structure including the absolute configuration was determined by spectroscopic data analysis and confirmed by X-ray crystallography. A plausible biosynthetic pathway is discussed on the basis of amino acid derivatization. Compound 1 showed inhibitory activity against histone deacetylase 1.