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Frontiers in Microbiology 2022spp. are traditional medicinal and edible filamentous fungi in China, and can produce various secondary metabolites, such as pigments (MPs) and citrinin (CIT). Genetic...
spp. are traditional medicinal and edible filamentous fungi in China, and can produce various secondary metabolites, such as pigments (MPs) and citrinin (CIT). Genetic modification methods, such as gene knock-out, complementation, and overexpression, have been used extensively to investigate the function of related genes in spp.. However, the resistance selection genes that can have been used for genetic modification in spp. are limited, and the gene replacement frequency (GRF) is usually <5%. Therefore, we are committed to construct a highly efficient gene editing system without resistance selection marker gene. In this study, using M7 as the starting strain, we successfully constructed a so-called markerlessly and highly genetic modification system including the mutants ΔΔ and ΔΔ::, in which we used the endogenous gene from M7 instead of the resistance marker gene as the screening marker, and simultaneously deleted related to non-homologous end joining in M7. Then, the morphology, the growth rate, the production of MPs and CIT of the mutants were analyzed. And the results show that the mutant strains have normal mycelia, cleistothecia and conidia on PDA+Uridine(U) plate, the biomass of each mutant is also no different from M7. However, the U addition also has a certain effect on the orange and red pigments yield of M7, which needs our further study. Finally, we applied the system to delete multiple genes from M7 separately or continuously without any resistance marker gene, and found that the average GRF of ΔΔ was about 18 times of that of M7. The markerlessly and highly genetic modification system constructed in current study not only will be used for multi-gene simultaneous modification in spp., and also lays a foundation for investigating the effects of multi-genes modification on spp..
PubMed: 35979480
DOI: 10.3389/fmicb.2022.952323 -
Journal of Fungi (Basel, Switzerland) Aug 2020pigments (MPs) have been used as food colorants for several centuries in Asian countries and are now used throughout the world via Asian catering. The MP biosynthetic...
pigments (MPs) have been used as food colorants for several centuries in Asian countries and are now used throughout the world via Asian catering. The MP biosynthetic pathway has been well-illustrated, but the functions of a few genes, including , in the MP gene cluster are still unclear. In the current study, in order to investigate the function of in M7, gene deletion (Δ), complementation (Δ::) and overexpression (M7::-) mutants were successfully obtained. The morphologies and biomasses, as well as the MP and citrinin production, of these mutants were analyzed. The results revealed that the disruption, complementation and overexpression of showed no apparent defects in morphology, biomass or citrinin production (except MP production) in Δ compared with M7. Although the MP profiles of Δ and M7 were almost the same-with both having four yellow pigments, two orange pigments (OPs) and two red pigments (RPs)-their yields were decreased in Δ to a certain extent. Particularly, the content of rubropunctatin (an OP) and its derivative rubropunctamine (an RP) in Δ, both of which have a five-carbon side chain, accounted for 57.7%, and 22.3% of those in M7. On the other hand, monascorubrin (an OP) and its derivative monascorubramine (an RP), both of which have a seven-carbon side chain, were increased by 1.15 and 2.55 times, respectively, in Δ compared with M7. These results suggest that the MrPigG protein may preferentially catalyze the biosynthesis of MPs with a five-carbon side chain.
PubMed: 32872515
DOI: 10.3390/jof6030156 -
Frontiers in Microbiology 2020It is well known that many organisms can perceive the magnetic field (MF), including the geomagnetic field, but how to feel MF is unclear. Recently, a study has claimed...
It is well known that many organisms can perceive the magnetic field (MF), including the geomagnetic field, but how to feel MF is unclear. Recently, a study has claimed that a biological compass, namely a complex of the magnetic receptor (MagR) and blue light (BL) receptor (cryptochrome), has been found in , , and , which may bring some new ideas to explore the mechanism of biomagnetism. spp. are edible filamentous fungi that can produce abundant beneficial secondary metabolites and have been used to produce food colorants for nearly 2000 years in the world, especially in China, Japan, and Korea. In this work, we firstly treated M7 by BL (500 lux,465-467 nm), MF (5, 10, 30 mT), and the combination of MF and BL (MF-BL), respectively. The results revealed that, compared with the control (CK, neither BL nor MF), the MF alone had no effect on the growth and morphological characteristics of M7, but BL made the colonial diameters only 66.7% of CK's and inhibited the formation of cleistothecia. Under MF-BL, the colony diameters were still 66.7% of CK's, but the colonial growth and cleistothecia production inhibited by BL were partially restored. Then, we have found that the gene widely exists in the genomes of animals, plants, and microorganisms, and we have also discovered a gene in the M7 genome, hereinafter referred to . Finally, the full-length cDNA of was successfully cloned and expressed in BL21 (DE3), and the Mr-MagR protein was purified by a Ni-NTA column and identified by Western blot. These results have laid a foundation for further investigation on the relationship between Mr-MagR and BL receptor(s) that might exist in M7. According to a literature search, it is the first time to report in filamentous fungi.
PubMed: 32636810
DOI: 10.3389/fmicb.2020.01112 -
Molecules (Basel, Switzerland) Dec 2018Yellow pigments can be of two kinds: Natural and reduced, in which natural yellow pigments (NYMPs) attract widespread attention for their bioactivities. In this study,...
Yellow pigments can be of two kinds: Natural and reduced, in which natural yellow pigments (NYMPs) attract widespread attention for their bioactivities. In this study, the antioxidative and antibreast cancer effects of the water-soluble NYMPs fermented by CGMCC 10910 were evaluated. Results showed that water-soluble NYMPs had a significantly improved antioxidative activities compared to the reduced yellow pigments (RYMPs) that were chemically derived from orange or red pigments. Furthermore, NYMPs exhibited a concentration-dependent inhibition activity on MCF-7 cell growth ( < 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.
Topics: Antineoplastic Agents; Antioxidants; Benzothiazoles; Biphenyl Compounds; Cell Death; Cell Movement; Cell Proliferation; Fermentation; Free Radical Scavengers; Humans; MCF-7 Cells; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Monascus; Neoplasm Invasiveness; Picrates; Pigments, Biological; Solubility; Sulfonic Acids; Vascular Endothelial Growth Factor A; Water
PubMed: 30544614
DOI: 10.3390/molecules23123242 -
Journal of Food and Drug Analysis Mar 2023Red mold rice (RMR) is a traditional Chinese medicine prepared using Monascus fermentation. Monascus ruber ( pilosus) and Monascus purpureus have a long history of use...
Red mold rice (RMR) is a traditional Chinese medicine prepared using Monascus fermentation. Monascus ruber ( pilosus) and Monascus purpureus have a long history of use as food and medicine. As an economically important starter culture, the relationship between the taxonomy of Monascus and production capabilities of secondary metabolites is crucial for the Monascus food industry. In this study, monacolin K, monascin, ankaflavin, and citrinin production by M. purpureus and M. ruber were genomically and chemically investigated. Our findings suggest that M. purpureus can produce monascin and ankaflavin in a correlated manner, whereas M. ruber produces monascin with minimum ankaflavin. M. purpureus is capable of producing citrinin; however, it is unlikely able to produce monacolin K. In contrast, M. ruber produces monacolin K, but not citrinin. We suggest that the current monacolin K content-related regulation of Monascus food should be revised, and labeling of Monascus species should be considered.
Topics: Monascus; Fermentation; Lovastatin; Oryza
PubMed: 37224553
DOI: 10.38212/2224-6614.3438 -
3 Biotech Feb 2018The present study verified whether acyl-coenzyme A (acyl-CoA)-binding protein (ACBP) affected the production of pigments (MPs) in CICC41233 (MrACBP). Phylogenetic...
The present study verified whether acyl-coenzyme A (acyl-CoA)-binding protein (ACBP) affected the production of pigments (MPs) in CICC41233 (MrACBP). Phylogenetic analysis revealed that the cloned gene, which encoded the MrACBP protein, exhibited the closest match (99% confidence level) to the gene from . The MrACBP and maltose-binding protein (MBP) were simultaneously expressed in Rosetta DE3 in the form of a fusion protein. The microscale thermophoresis binding assay revealed that the purified MBP-MrACBP exhibited a higher affinity for myristoyl-CoA (Kd = 88.16 nM) than for palmitoyl-CoA (Kd = 136.07 nM) and octanoyl-CoA (Kd = 270.9 nM). Further, the gene was homologously overexpressed in CICC41233, and a positive transformant ACBP5 was isolated. The fatty acid myristic acid in ACBP5 was lower than that in the parent strain CICC41233. However, when compared with the parent strain, the production of total MPs, water-soluble pigment, and ethanol-soluble pigment in ACBP5 increased by 11.67, 9.80, and 12.70%, respectively, after 6 days. The relative gene expression level, as determined by a quantitative real-time polymerase chain reaction analysis, of the key genes , , , , and increased by 4.03-, 3.58-, 1.67-, 2.11-, and 2.62-fold after 6 days. These data demonstrate the binding preference of MrACBP for myristoyl-CoA, and its influence on MPs production.
PubMed: 29430382
DOI: 10.1007/s13205-018-1147-9 -
Journal of Fungi (Basel, Switzerland) Dec 2021pigments (MPs) have been used as food colorants for several centuries in Asian countries and are currently used around the world via Asian catering. The MPs...
pigments (MPs) have been used as food colorants for several centuries in Asian countries and are currently used around the world via Asian catering. The MPs biosynthetic pathway has been well-illustrated; however, the functions of a few genes including in the MPs gene cluster of M7 are still unclear. In the current study, was disrupted in ΔΔ, a highly efficient gene modification system, using as a selection marker, and ΔΔΔ:: and ΔΔΔ have been obtained. Subsequently, their morphologies, biomasses, MPs and citrinin (CIT) production were analyzed, respectively. These results have revealed that the deletion of has significant effects on the morphology and growth of M7. Moreover, compared with M7, the yields of MPs and CIT were drastically increased and decreased in mutants, respectively.
PubMed: 34947076
DOI: 10.3390/jof7121094 -
Brazilian Journal of Microbiology :... Jun 2021Three culture media were studied for red pigment production by Monascus ruber in submerged cultivation: rice flour (20 g L), sugarcane molasses (30 g L), and, finally,...
Three culture media were studied for red pigment production by Monascus ruber in submerged cultivation: rice flour (20 g L), sugarcane molasses (30 g L), and, finally, molasses + rice flour (10 g L+10 g L); all culture media were added of 5 g L glycine as nitrogen source. Rice flour showed pigment production of 7.05 UA and molasses 5.08 UA, and the mixture of rice flour and molasses showed the best result of 16.38 UA Molasses culture presented good results for cell biomass production of 11.09 g L. With these results, it was observed that one substrate presented good pigment production (rice flour) and another attained better results for cell biomass growth (molasses), and a third medium containing 10 g L of rice flour + 10 g L of molasses was formulated. The results for this mixture showed satisfactory results, with global pigment productivity of 0.097 UA h and maximum productivity rate of 0.17 UA h. The high production and productivity obtained for the mixture of rice flour and molasses indicated that the production of red pigment by submerged fermentation, using the mixture of these low-cost culture media, may be promising in terms of commercial production.
Topics: Biotransformation; Fermentation; Flour; Molasses; Monascus; Oryza; Pigments, Biological; Saccharum; Waste Products
PubMed: 33651332
DOI: 10.1007/s42770-021-00456-9 -
Frontiers in Microbiology 2021Cocultivation is an emerging and potential way to investigate microbial interaction in the laboratory. Extensive researches has been carried out over the years, but some...
Cocultivation is an emerging and potential way to investigate microbial interaction in the laboratory. Extensive researches has been carried out over the years, but some microorganism cocultivation are not easy to implement in the laboratory, especially the fungus-fungus (FF) cocultivation, owing to the obstacles such as fungal different growth rate, limited growing space, hyphae intertwining, and difficulty of sample separation, etc. In this research, a double-sided petri dish (DSPD) was designed and carried out as a tool to study FF cocultivation in the laboratory. A natural FF cocultivation of spp. and inspired from black-skin-red-koji (BSRK), were studied. By using DSPD, the aforementioned obstacles in the FF cocultivation study were overcome through co-culturing spp. and on each side of DSPD. The characteristics of monocultured and co-cultured spp. and were compared and analyzed, including colonial and microscopic morphologies, and main secondary metabolites (SMs) of spp. analyzed by high performance liquid chromatography. And a novel SM was found to be produced by M7 when co-cultured with CBS 513.88. Since the above mentioned obstacles, were overcome, we obtained good quality of transcriptome data for further analysis. These results indicate that DSPD might be an efficient tool for investigation of microbial interaction, in particular, for FF interaction.
PubMed: 34177849
DOI: 10.3389/fmicb.2021.670684 -
Marine Drugs Feb 2020Three new compounds, monarubins A-C (, and ), together with ten known compounds, including four alkaloids (-), two isocoumarins ( and ) and four polyketides (-), were...
Three new compounds, monarubins A-C (, and ), together with ten known compounds, including four alkaloids (-), two isocoumarins ( and ) and four polyketides (-), were isolated from marine shellfish-associated fungus BB5. The structures were determined on the basis of the 1D and 2D NMR, MS, UV and IR data. The absolute configurations of compounds , and were determined by ECD calculations. The NMR data of compounds deoxyhydroxyaspergillic acid () and 2-hydroxy-6-(1-hydroxy-1-methylpropyl)-3--buthylpyrazine () were first reported. All of the isolated compounds were evaluated for their cytotoxic activities against human nasopharyngeal carcinoma cell lines CNE1, CNE2, SUNE1 and HONE1 and hepatocellular carcinoma cell lines QGY7701 and HepG2. Monarubin B () displayed potent cytotoxicities against the cancer cell lines HepG2 and QGY7701 with IC values of 1.72 and 0.71 μΜ, respectively; lunatinin () showed moderate cytotoxic activities against the cancer cell lines HepG2, QGY7701 and SUNE1 with the IC values of 9.60, 7.12 and 28.12 μΜ, respectively.
Topics: Alkaloids; Animals; Antineoplastic Agents; Carcinoma, Hepatocellular; Cell Line, Tumor; Hep G2 Cells; Humans; Isocoumarins; Liver Neoplasms; Monascus; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Polyketides; Shellfish
PubMed: 32028626
DOI: 10.3390/md18020100