Biotransformation of hydrocortisone succinate with whole cell cultures of Monascus purpureus and Cunninghamella echinulate.

Hydrocortisone succinate (1) is a synthetic anti-inflammatory drug and key intermediate in the synthesis of other steroidal drugs. This work is based on the fungal biotransformation of 1, using Monascus purpureus and Cunninghamella echinulata strains. Comopound 1 was transformed into four metabolites, identified as hydrocortisone (2), 11ß-hydroxyandrost-4-en-3,17-dione (3), Δ1-cortienic acid (4), and hydrocortisone-17-succinate (5), obtained through side chain cleavage, hydrolysis, dehydrogenation, and oxidation reactions. These compounds have previously been synthesized either chemically or enzymatically from different precursors. Though this is not the first report on the biotransformation of 1, but it obviously is a first, where the biotransformed products of compound 1 have been characterized structurally with the help of modern spectroscopic techniques. It is noteworthy that these products have already shown biological potential, however a more thorough investigation of the anti-inflammatory properties of these metabolites would be of high value. These results not only emphasize upon the immense potential of biotransformation in catalysis of reactions, otherwise not-achievable chemically, but also holds promise for the development of novel anti-inflammatory compounds.

Insight into selenium biofortification and the selenite metabolic mechanism of Monascus ruber M7.

Monascus species are functional fermentation fungi with great potential for selenium (Se) supplementation. This study investigated the effects of Se bio-fortification on the growth, morphology, and biosynthesis of Monascus ruber M7. The results demonstrated a significant increase in the yield of orange and red Monascus pigments (MPs) in red yeast rice (RYR) by 38.52% and 36.57%, respectively, under 20 µg/mL of selenite pressure. Meanwhile, the production of citrinin (CIT), a mycotoxin, decreased from 244.47 µg/g to 175.01 µg/g. Transcriptome analysis revealed significant upregulation of twelve genes involved in MPs biosynthesis, specifically MpigE, MpigF, and MpigN, and downregulation of four genes (mrr3, mrr4, mrr7, and mrr8) associated with CIT biosynthesis. Additionally, three genes encoding cysteine synthase cysK (Log2FC = 1.6), methionine synthase metH (Log2FC = 2.2), and methionyl-tRNA synthetase metG (Log2FC = 1.8) in selenocompound metabolism showed significantly upregulated. These findings provide insights into Se biotransformation and metabolism in filamentous fungi.

Unlocking fungal quorum sensing: Oxylipins and yeast interactions enhance secondary metabolism in monascus.

Exploring the symbiotic potential between fungal and yeast species, this study investigates the co-cultivation dynamics of Monascus, a prolific producer of pharmacologically relevant secondary metabolites, and Wickerhamomyce anomalous. The collaborative interaction between these microorganisms catalyzed a substantial elevation in the biosynthesis of secondary metabolites, prominently Monacolin K and natural pigments. Central to our discoveries was the identification and enhanced production of oxylipins (13S-hydroxyoctadecadienoic acid，13S-HODE), putative quorum-sensing molecules, within the co-culture environment. Augmentation with exogenous oxylipins not only boosted Monacolin K production by over half but also mirrored morphological adaptations in Monascus, affecting both spores and mycelial structures. This augmentation was paralleled by a significant upregulation in the transcriptional activity of genes integral to the Monacolin K biosynthetic pathway, as well as genes implicated in pigment and spore formation. Through elucidating the interconnected roles of quorum sensing, G-protein-coupled receptors, and the G-protein-mediate signaling pathway, this study provides a comprehensive view of the molecular underpinnings facilitating these metabolic enhancements. Collectively, our findings illuminate the profound influence of Wickerhamomyces anomalous co-culture on Monascus purpureus, advocating for oxylipins as a pivotal quorum-sensing mechanism driving the observed symbiotic benefits.

Novel compounds isolated from health food products containing beni-koji (red yeast rice) with adverse event reports.

Recently, health hazards, such as kidney damage, have been reported owing to the ingestion of a health food product, so-called "foods with functional claims (FFC)'', containing beni-koji (red yeast rice). Although not an expected compound in the FFC, the detection of puberulic acid has also been reported. Further investigations of these health food products, such as the identification of other unintended compounds and clarifying the health impacts of puberulic acid, are required. To clarify the causes of these health issues, we investigated the presence of unintended compounds in the FFC containing beni-koji using comprehensive instrumental analyses. Using differential analysis, novel compounds 1 and 2 were detected as unexpected components between the samples with and without adverse event reports. Although limited to the samples available for analyses in this study, both compounds 1 and 2 were detected in all the samples that also contained puberulic acid. Compounds 1 and 2, with molecular formulas of C23H34O7 and C28H42O8, respectively, may be lovastatin derivatives. Their structures were confirmed using NMR analyses and are novel natural compounds. For definitive confirmation, we are in the process of synthesizing compounds 1 and 2 from lovastatin. The route of contamination of these compounds are currently under investigation. The findings of this study could be used to address the growing health hazards associated with health food products.

Metagenomic and flavoromic profiling reveals the correlation between the microorganisms and volatile flavor compounds in Monascus-fermented cheese.

The Monascus-fermented cheese (MC) is a unique cheese product that undergoes multi-strain fermentation, imparting it with distinct flavor qualities. To clarify the role of microorganisms in the formation of flavor in MC, this study employed SPME (arrow)-GC-MS, GC-O integrated with PLS-DA to investigate variations in cheese flavors represented by volatile flavor compounds across 90-day ripening periods. Metagenomic datasets were utilized to identify taxonomic and functional changes in the microorganisms. The results showed a total of 26 characteristic flavor compounds in MC at different ripening periods (VIP＞1, p < 0.05), including butanoic acid, hexanoic acid, butanoic acid ethyl ester, hexanoic acid butyl ester, 2-heptanone and 2-octanone. According to NR database annotation, the genera Monascus, Lactococcus, Aspergillus, Lactiplantibacillus, Staphylococcus, Flavobacterium, Bacillus, Clostridium, Meyerozyma, and Enterobacter were closely associated with flavor formation in MC. Ester compounds were linked to Monascus, Meyerozyma, Staphylococcus, Lactiplantibacillus, and Bacillus. Acid compounds were linked to Lactococcus, Lactobacillus, Staphylococcus, and Bacillus. The production of methyl ketones was closely related to the genera Monascus, Staphylococcus, Lactiplantibacillus, Lactococcus, Bacillus, and Flavobacterium. This study offers insights into the microorganisms of MC and its contribution to flavor development, thereby enriching our understanding of this fascinating dairy product.

Exploring the Subcellular Localization of Monascus Pigments Biosynthases: Preliminary Unraveling of the Compartmentalization Mechanism.

Monascus pigments (MPs), a class of secondary metabolites produced by Monascus spp., can be classified into yellow, orange, and red MPs according to their differences in the wavelength of the maximum absorption. However, the biosynthetic sequence and cellular biosynthesis mechanism of different MPs components are still not yet completely clear in Monascus spp. In this study, the subcellular localization of five MPs synthases was investigated using fluorescent protein fusion expression. The results revealed that the proteins encoded by the MPs biosynthetic gene cluster were compartmentalized in various subcellular locations, including the mitochondrial polyketide synthase MrPigA, cytosolic enzymes consisting of the ketoreductase MrPigC, the oxidoreductase MrPigE, and the monooxygenase MrPigN, and the cell-wall-bound oxidoreductase MrPigF. Moreover, the correct localization of MrPigF to the cell wall was crucial for the synthesis of orange MPs. Lastly, we discussed the compartmentalized biosynthetic pathway of MPs. This study will not only be helpful in clarifying the biosynthetic sequence and biosynthesis mechanism of different MPs but also provides new insights into the cellular biosynthesis of secondary metabolites in filamentous fungi.

Taurine-mediated gene transcription and cell membrane permeability reinforced co-production of bioethanol and Monascus azaphilone pigments for a newly isolated Monascus purpureus.

BACKGROUND: Taurine, a semi-essential micronutrient, could be utilized as a sulfur source for some bacteria; however, little is known about its effect on the accumulation of fermentation products. Here, it investigated the effect of taurine on co-production of bioethanol and Monascus azaphilone pigments (MonAzPs) for a fungus. RESULTS: A newly isolated fungus of 98.92% identity with Monascus purpureus co-produced 23.43 g/L bioethanol and 66.12, 78.01 and 62.37 U/mL red, yellow and orange MonAzPs for 3 d in synthetic medium (SM). Taurine enhanced bioethanol titer, ethanol productivity and ethanol yield at the maximum by 1.56, 1.58 and 1.60 times than those of the control in corn stover hydrolysates (CSH), and red, yellow and orange MonAzPs were raised by 1.24, 1.26 and 1.29 times, respectively. Taurine was consumed extremely small quantities for M. purpureus and its promotional effect was not universal for the other two biorefinery fermenting strains. Taurine intensified the gene transcription of glycolysis (glucokinase, phosphoglycerate mutase, enolase and alcohol dehydrogenase) and MonAzPs biosynthesis (serine hydrolases, C-11-ketoreductase, FAD-dependent monooxygenase, 4-O-acyltransferase, deacetylase, NAD(P)H-dependent oxidoredutase, FAD-dependent oxidoredutase, enoyl reductase and fatty acid synthase) through de novo RNA-Seq assays. Furthermore, taurine improved cell membrane permeability through changing cell membrane structure by microscopic imaging assays. CONCLUSIONS: Taurine reinforced co-production of bioethanol and MonAzPs by increasing gene transcription level and cell membrane permeability for M. purpureus. This work would offer an innovative, efficient and taurine-based co-production system for mass accumulation of the value-added biofuels and biochemicals from lignocellulosic biomass.

Research on pear residue dietary fiber and Monascus pigments extracted through liquid fermentation.

Pear residue, a byproduct of pear juice extraction, is rich in soluble sugar, vitamins, minerals, and cellulose. This study utilized Monascus anka in liquid fermentation to extract dietary fiber (DF) from pear residue, and the structural and functional characteristics of the DF were analyzed. Soluble DF (SDF) content was increased from 7.9/100 g to 12.6 g/100 g, with a reduction of average particle size from 532.4 to 383.0 nm by fermenting with M. anka. Scanning electron microscopy and infrared spectroscopic analysis revealed more porous and looser structures in Monascus pear residue DF (MPDF). Water-, oil-holding, and swelling capacities of MPDF were also enhanced. UV-visible spectral analysis showed that the yield of yellow pigment in Monascus pear residue fermentation broth (MPFB) was slightly higher than that in the Monascus blank control fermentation broth. The citrinin content in MPFB and M. anka seed broth was 0.90 and 0.98 ug/mL, respectively. Therefore, liquid fermentation with M. anka improved the structural and functional properties of MPDF, suggesting its potential as a functional ingredient in food.

Effects of MrwetA on Sexual Reproduction and Secondary Metabolism of Monascus ruber M7 Based on Transcriptome Analysis.

wetA, one of the conidiation center regulatory genes in many filamentous fungi, plays an important role in promoting asexual spores (conidia) maturation. Our recent research has found that knocking out or overexpressing MrwetA (a homolog of wetA) in Monascus ruber M7 does not affect the development of its asexual spores like other fungi, but both repress the development of its sexual spores (ascospores). However, the mechanism remains unclear. In this study, the function of MrwetA on sexual reproduction and secondary metabolism in M. ruber M7 was confirmed by a complementary experiment. Moreover, the regulatory roles of MrwetA in modulating the expression of genes involved in sexual reproduction, meiosis, and biosynthesis of Monascus pigment and citrinin were analyzed based on the transcriptional data. These results not only contribute to clarifying the regulation of the reproduction and secondary metabolism of Monascus spp., but also to enriching the regulation molecular mechanism of reproduction in filamentous fungi.

Monascus purpureus M-32 fermented soybean meal improves the growth, immunity parameters, intestinal morphology, disease resistance, intestinal microbiota and metabolome in Pacific white shrimp (Litopenaeus vannamei).

This study was conducted to evaluate the effects of Monascus purpureus M-32 fermented soybean meal (MFSM) on growth, immunity, intestinal morphology, intestinal microbiota, and intestinal metabolome of Pacific white shrimp (Litopenaeus vannamei). Four groups of diets were formulated, including control group (30% fish meal and 30% soybean meal [SBM] included in the basal diet) and three experimental groups which MFSM replaced 20% (MFSM20), 40% (MFSM40), and 60% (MFSM60) of SBM in control group, respectively. Results showed that the soluble proteins larger than 49 kDa in MFSM were almost completely degraded. Meanwhile, the crude protein, acid-soluble protein, and amino acid in MFSM were increased. The results of shrimp culture experiment showed that the replacement of SBM with MFSM decreased FCR (P < 0.001) and content of malondialdehyde (P = 0.007) in the experimental groups, and increased weight gain rate (P = 0.006), specific growth rate (P = 0.002), survival rate (P = 0.005), intestinal villus height (P < 0.001), myenteric thickness (P = 0.002), the activities of superoxide dismutase (P = 0.002), and lysozyme (P = 0.006) in experimental groups, as well as increased content of calcium (Ca2+) and phosphorus (PO43-) in blood and muscle, and enhanced resistance to Vibrio parahaemolyticus infection. The gut microbiota of MFSM groups was significantly different from that of the control group, and the abundance of Actinobacteria and Verrucomicrobia increased significantly in the MFSM60 group, whereas Proteobacteria and Firmicutes decreased. Compared with the control group, there were significant changes in the levels of several intestinal metabolites in the MFSM60 group, including leukotriene C5, prostaglandin A1, taurochenodeoxycholic acid, carnosine, and itaconic acid. The fermentation of SBM by the strain M. purpureus M-32 has the potential to enhance the nutritional quality of SBM, promote the growth of L. vannamei, boost immune response, improve intestinal morphology and microbiota composition, as well as influence intestinal metabolites.

Enhancing extracellular monascus pigment production in submerged fermentation with engineered microbial consortia.

In this study, we investigated the impact of microbial interactions on Monascus pigment (MP) production. We established diverse microbial consortia involving Monascus purpureus and Lactobacillus fermentum. The addition of Lactobacillus fermentum (4% at 48 h) to the submerged fermentation of M. purpureus resulted in a significantly higher MP production compared to that achieved using the single-fermentation system. Co-cultivation with immobilized L. fermentum led to a remarkable increase of 59.18% in extracellular MP production, while mixed fermentation with free L. fermentum caused a significant decrease of 66.93% in intracellular MPs, contrasting with a marginal increase of 4.52% observed during co-cultivation with immobilized L. fermentum and the control group respectively. The findings indicate an evident enhancement in cell membrane permeability of M. purpureus when co-cultivated with immobilized L. fementum. Moreover, integrated transcriptomic and metabolomic analyses were conducted to elucidate the regulatory mechanisms underlying MP biosynthesis and secretion following inoculation with immobilized L. fementum, with specific emphasis on glycolysis, steroid biosynthesis, fatty acid biosynthesis, and energy metabolism.

Stability mechanism of Monascus pigment-soy protein isolate-maltodextrin complex.

BACKGROUND: Monascus pigment (MP) is a natural food coloring with vital physiological functions but prone to degradation and color fading under light conditions. RESULTS: This study investigated the effect of complex formation of soybean protein isolate (SPI), maltodextrin (MD), and MP on the photostability of MP. Light stability was assessed through retention rate and color difference. Fluorescence spectroscopy (FS), circular dichroism (CD), Fourier-transform infrared (FTIR) spectroscopy, and X-ray diffraction (XRD) explored MP, SPI, and MD interactions, clarifying the MP-SPI-MD complex mechanism on the light stability of MP. Microstructure and differential scanning calorimetry (DSC) analyzed the morphology and thermal properties. The retention rate of MP increased to approximately 80%, and minimal color difference was observed when adding SPI and MD simultaneously. FS revealed hydrophobic interaction between MP and SPI. FTIR analysis showed intensity changes and peak shifts in amide I band and amide II band, which proved the hydrophobic interaction. CD showed a decrease in α-helix content and an increase in ß-sheet content after complex formation, indicating strengthened hydrogen bonding interactions. Scanning electron microscopy (SEM) analysis demonstrated that MP was attached to the surface and interior of complexes. XRD showed MP as crystalline, while SPI and MD were amorphous, complexes exhibited weakened or absent peaks, suggesting MP encapsulation. The results of DSC were consistent with XRD. CONCLUSION: SPI and MD enveloped MP through hydrogen bonding and hydrophobic interaction, ultimately enhancing its light stability and providing insights for pigment-protein-polysaccharide interactions and improving pigment stability in the food industry. © 2024 Society of Chemical Industry.

Improving Soluble Phenolic Profile and Antioxidant Activity of Grape Pomace Seeds through Fungal Solid-State Fermentation.

Grape pomace seeds contain abundant phenolic compounds, which are also present in both soluble and insoluble forms, similar to many other plant matrices. To further increase the extractable soluble phenolics and their antioxidant activities, grape pomace seeds were fermented with different fungi. Results showed that solid-state fermentation (SSF) with Aspergillus niger, Monascus anka, and Eurotium cristatum at 28 °C and 65% humidity had a significantly positive impact on the release of soluble phenolics in grape pomace seeds. Specifically, SSF with M. anka increased the soluble phenolic contents by 6.42 times (calculated as total phenolic content) and 6.68 times (calculated as total flavonoid content), leading to an overall improvement of antioxidant activities, including DPPH (increased by 2.14 times) and ABTS (increased by 3.64 times) radical scavenging activity. Furthermore, substantial changes were observed in the composition and content of individual phenolic compounds in the soluble fraction, with significantly heightened levels of specific phenolics such as chlorogenic acid, syringic acid, ferulic acid, epicatechin gallate, and resveratrol. Notably, during M. anka SSF, positive correlations were identified between the soluble phenolic content and hydrolase activities. In particular, there is a strong positive correlation between glycosidase and soluble phenols (r = 0.900). The findings present an effective strategy for improving the soluble phenolic profiles and bioactivities of grape pomace seeds through fungal SSF, thereby facilitating the valorization of winemaking by-products.

The effects of environmental factors on the synthesis of water-soluble Monascus red pigments via submerged fermentation: a review.

Monascus pigments (MPs) have been used as natural food pigments for many years. There is a high demand for Monascus red pigments (MRPs) to enhance color and for antibacterial and cancer prevention therapies in food and medicine. Most MRPs are not water soluble, and the yield of water-soluble MRPs is naturally low. On the other hand, water-soluble MRP is more cost effective for application in industrial mass production. Therefore, it is important to improve the yield of water-soluble MRPs. Environmental factors have a significant influence on the synthesis of water-soluble MRPs, which is crucial for the development of industrial production of water-soluble MRPs. This review introduces the biosynthetic pathways of water-soluble MRPs and summarizes the effects of environmental factors on the yield of water-soluble MRPs. Acetyl coenzyme A (acetyl-CoA) is a precursor for MPs synthesis. Carbon and nitrogen sources and the carbon/nitrogen ratio can impact MP production by regulating the metabolic pathway of acetyl-CoA. Optimization of fermentation conditions to change the morphology of Monascus can stimulate the synthesis of MPs. The appropriate choice of nitrogen sources and pH values can promote the synthesis of MRPs from MPs. Additives such as metal ions and non-ionic surfactants can affect the fluidity of Monascus cell membrane and promote the transformation of MRPs into water-soluble MRPs. This review will lay the foundation for the industrial production of water-soluble MRPs. © 2024 Society of Chemical Industry.

Occurrence, Risk Implications, Prevention and Control of CIT in Monascus Cheese: A Review.

Monascus is a filamentous fungus that has been used in the food and pharmaceutical industries. When used as an auxiliary fermenting agent in the manufacturing of cheese, Monascus cheese is obtained. Citrinin (CIT) is a well-known hepatorenal toxin produced by Monascus that can harm the kidneys structurally and functionally and is frequently found in foods. However, CIT contamination in Monascus cheese is exacerbated by the metabolic ability of Monascus to product CIT, which is not lost during fermentation, and by the threat of contamination by Penicillium spp. that may be introduced during production and processing. Considering the safety of consumption and subsequent industrial development, the CIT contamination of Monascus cheese products needs to be addressed. This review aimed to examine its occurrence in Monascus cheese, risk implications, traditional control strategies, and new research advances in prevention and control to guide the application of biotechnology in the control of CIT contamination, providing more possibilities for the application of Monascus in the cheese industry.

An Exopolysaccharide from Genistein-Stimulated Monascus Purpureus: Structural Characterization and Protective Effects against DSS-Induced Intestinal Barrier Injury Associated with the Gut Microbiota-Modulated Short-Chain Fatty Acid-TLR4/MAPK/NF-κB Cascade Response.

Inflammatory bowel disease is a major health problem that can lead to prolonged damage to the digestive system. This study investigated the effects of an exopolysaccharide from genistein-stimulated Monascus purpureus (G-EMP) in a mouse model of colitis to clarify its molecular mechanisms and identified its structures. G-EMP (Mw = 56.4 kDa) was primarily consisted of → 4)-α-D-Galp-(1 →, → 2,6)-α-D-Glcp-(1→ and →2)-ß-D-Manp-(1 → , with one of the branches being α-D-Manp-(1 →. G-EMP intervention reduced the loss of body weight, degree of colonic damage and shortening, disease activity index scores, and histopathology scores, while restoring goblet cell production and oxidative homeostasis, repairing colonic functions, and regulating inflammatory cytokines. RNA sequencing and Western blot analysis indicated that G-EMP exerts anti-inflammatory properties by suppressing the TLR4/MAPK/NF-κB inflammatory signaling pathway. G-EMP modulated the gut microbiota by improving its diversities, elevating the relative abundances of beneficial bacteria, declining the Firmicutes/Bacteroidota value, and regulating the level of short-chain fatty acids (SCFAs). Correlation analysis demonstrated strong links between SCFAs, gut microbiota, and the inflammatory response, indicating the potential of G-EMP to prevent colitis.

COMPASS core subunits MpSet1 and MpSwd3 regulate Monascus pigments synthesis in Monascus purpureus.

In eukaryotes, methylation of histone H3 at lysine 4 (H3K4me) catalyzed by the complex of proteins associated with Set1 (COMPASS) is crucial for the transcriptional regulation of genes and the development of organisms. In Monascus, the functions of COMPASS in establishing H3K4me remain unclear. This study first identified the conserved COMPASS core subunits MpSet1 and MpSwd3 in Monascus purpureus and confirmed their roles in establishing H3K4me2/3. Loss of MpSet1 and MpSwd3 resulted in slower growth and development and inhibited the formation of cleistothecia, ascospores, and conidia. The loss of these core subunits also decreased the production of extracellular and intracellular Monascus pigments (MPs) by 94.2%, 93.5%, 82.7%, and 82.5%, respectively. In addition, RNA high-throughput sequencing and quantitative real-time polymerase chain reaction (qRT-PCR) showed that the loss of MpSet1 and MpSwd3 altered the expression of 2646 and 2659 genes, respectively, and repressed the transcription of MPs synthesis-related genes. In addition, the ΔMpset1 and ΔMpswd3 strains demonstrated increased sensitivity to cell wall stress with the downregulation of chitin synthase-coding genes. These results indicated that the COMPASS core subunits MpSet1 and MpSwd3 help establish H3K4me2/3 for growth and development, spore formation, and pigment synthesis in Monascus. These core subunits also assist in maintaining cell wall integrity.

Enhancement of yellow pigments production via high CaCl2 stress fermentation of Monascus purpureus.

Monascus pigments (MPs) are a kind of natural ingredient fermented by Monascus spp., which contains three types of pigments: red, orange, and yellow ones. Monascus yellow pigments have a restricted yield and cannot meet industrial application. The method and mechanism of CaCl2 improving yellow pigments production by liquid fermentation of Monascus purpureus M8 were studied in order to overcome the low yield of yellow pigments produced by liquid fermentation. Changes in physiological and biochemical indicators explained the effects of CaCl2 on the production of Monascus yellow pigments from solid fermentation. The intracellular yellow pigments, orange pigments, and red pigments increased by 156.08%, 43.76%, and 42.73%, respectively, with 60 g/l CaCl2 addition to culture medium. The amount of red and orange pigments reduced, while the proportion of yellow pigments increased and the relative peak area of intracellular yellow pigments accounted for a dominant 98.2%, according to thin layer chromatography and high performance liquid chromatography analyses. Furthermore, the influence of CaCl2 extended to the modulation of pigments synthesis-related gene expression in M8 strain. This modulation led to a pronounced upregulation in the expression of the yellow pigments synthesis-related gene, mppE, signifying a pivotal role played by CaCl2 in orchestrating the intricate machinery behind yellow pigments biosynthesis.

Identification of fungi isolated from commercial bumblebee colonies.

Fungi can have important beneficial and detrimental effects on animals, yet our understanding of the diversity and function of most bee-associated fungi is poor. Over 2 million bumblebee colonies are traded globally every year, but the presence and transport of viable fungi within them is unknown. Here, we explored whether any culturable fungi could be isolated from commercial bumblebee nests. We collected samples of various substrates from within 14 bumblebee colonies, including the honey, honey cup wall, egg cup wall, and frass then placed them on agar and recorded any growth. Fungal morphotypes were then subcultured and their ITS region sequenced for identification. Overall, we cultured 11 fungal species from the various nest substrates. These included both pathogenic and non-pathogenic fungi, such as Aspergillus sp., Penicillium sp., and Candida sp. Our results provide the first insights into the diversity of viable fungal communities in commercial bumblebee nests. Further research is needed to determine if these fungi are unique to commercial colonies or prevalent in wild bumblebee nests, and crucially to determine the ecological and evolutionary implications of these fungi in host colonies.

Disruption of UDP-galactopyranose mutase expression: A novel strategy for regulation of galactomannan biosynthesis and monascus pigments secretion in Monascus purpureus M9.

The impact of the cell wall structure of Monascus purpureus M9 on the secretion of extracellular monascus pigments (exMPs) was investigated. To modify the cell wall structure, UDP-galactopyranose mutase (GlfA) was knocked out using Agrobacterium-mediated transformation method, leading to a significant reduction in the Galf-based polysaccharide within the cell wall. Changes in mycelium morphology, sporogenesis, and the expression of relevant genes in M9 were also observed following the mutation. Regarding MPs secretion, a notable increase was observed in six types of exMPs (R1, R2, Y1, Y2, O1 and O2). Specifically, these exMPs exhibited enhancement of 1.33, 1.59, 0.8, 2.45, 2.89 and 4.03 times, respectively, compared to the wild-type strain. These findings suggest that the alteration of the cell wall structure could selectively influence the secretion of MPs in M9. The underlying mechanisms were also discussed. This research contributes new insights into the regulation of the synthesis and secretion of MPs in Monascus spp..