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Frontiers in Microbiology 2019Ypts (east rotein ransport),also called as as-ssociated inding GTPases (Rab), are the largest group of the small GTPases family, which have been extensively studied in...
Ypts (east rotein ransport),also called as as-ssociated inding GTPases (Rab), are the largest group of the small GTPases family, which have been extensively studied in model eukaryotic cells and play a pivotal role in membane trafficking, while this study showed potential regulation role of Ypts in fungi. One of Ypts, Ypt7 may be involved in fungal development and secondary metabolism, but the exact mechanism still exists a controversy. In current study, the functions of a homologous gene (7) from M7 was investigated by combination of gene-deletion (Δ7), overexpression (M7::-7) and transcriptome analysis. Results showed that the radial growth rate of Δ7 was significantly slower than M7, little conidia and ascospores can be observed in Δ7, but the yield of intracellular secondary metabolites was dramatically increased. Simultaneously, the 7 overexpression strain possessed similar capacity for sporulation and secondary metabolism observed in M7. Transcriptome results further illustrated that 7 could coordinate with numerous genes involved in the vegetative growth, conidiogenesis, secondary metabolism biosynthesis and transportation of M7. Combined with the similar effect of Ypt7 homologs on other fungi, we propose that Ypt7 works more like a global regulatory factor in fungi. To our knowledge, it is the first time to investigate Ypt7 functions in . It could also improve the understanding of Ypt7 functions in fungi.
PubMed: 30936855
DOI: 10.3389/fmicb.2019.00452 -
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 -
Studies in Mycology Sep 2018has long-been used as a model organism to gain insights into the genetic basis of asexual and sexual developmental processes both in other members of the genus , and...
has long-been used as a model organism to gain insights into the genetic basis of asexual and sexual developmental processes both in other members of the genus , and filamentous fungi in general. Paradigms have been established concerning the regulatory mechanisms of conidial development. However, recent studies have shown considerable genome divergence in the fungal kingdom, questioning the general applicability of findings from , and certain longstanding evolutionary theories have been questioned. The phylogenetic distribution of key regulatory elements of asexual reproduction in was investigated in a broad taxonomic range of fungi. This revealed that some proteins were well conserved in the ( AbaA, FlbA, FluG, NsdD, MedA, and some velvet proteins), suggesting similar developmental roles. However, other elements ( BrlA) had a more restricted distribution solely in the , and it appears that the genetic control of sporulation seems to be more complex in the aspergilli than in some other taxonomic groups of the . The evolution of the velvet protein family is discussed based on the history of expansion and contraction events in the early divergent fungi. Heterologous expression of the gene in failed to induce development of complete conidiophores as seen in the aspergilli, but did result in increased conidial production. The absence of many components of the asexual developmental pathway from members of the supports the hypothesis that differences in the complexity of their spore formation is due in part to the increased diversity of the sporulation machinery evident in the Pezizomycotina. Investigations were also made into the evolution of sex and sexuality in the aspergilli. loci were identified from the heterothallic () and () and the homothallic (=). A consistent architecture of the locus was seen in these and other heterothallic aspergilli whereas much variation was seen in the arrangement of loci in homothallic aspergilli. This suggested that it is most likely that the common ancestor of the aspergilli exhibited a heterothallic breeding system. Finally, the supposed prevalence of asexuality in the aspergilli was examined. Investigations were made using as a representative 'asexual' species. It was possible to induce a sexual cycle in given the correct and partners and environmental conditions, with recombination confirmed utilising molecular markers. This indicated that sexual reproduction might be possible in many supposedly asexual aspergilli and beyond, providing general insights into the nature of asexuality in fungi.
PubMed: 30425416
DOI: 10.1016/j.simyco.2018.10.002 -
3 Biotech Aug 2018To investigate the relationship between starch hydrolysis and pigments (MPs) production, the α-amylase gene () from was heterologously expressed in CICC41233, and we...
To investigate the relationship between starch hydrolysis and pigments (MPs) production, the α-amylase gene () from was heterologously expressed in CICC41233, and we obtained a positive transformant named Amy9. In Amy9, the α-amylase activities were 6.65- and 4.26-fold higher at 72 h and 144 h, respectively, than those in the parent strain with the glucose as solo carbon medium. Surprisingly, in the MPs fermentation medium with rice powder as solo material, Amy9 completely degraded starch at 48 h, while 43.93 and 7.29 mg/mL starch remained at 48 and 144 h, respectively, in the parent strain. Amy9 accelerated starch hydrolysis, which enhanced biomass and also increased total MPs by 132% after 144 h. Compared with CICC41233, the relative gene expression levels, as determined by a quantitative real-time polymerase chain reaction analysis, of encoding ATP-citrate lyase subunit 2, encoding polyketide synthase, and encoding the fatty acid synthase beta subunit increased by 33.14, 145.18, and 32.15%, respectively, after 144 h in Amy9. The up-regulated expression of these key genes in MPs synthesis contributed to the large increase in MPs production. This interesting work provided us with a new idea and a new target for the study of the MPs production.
PubMed: 30073114
DOI: 10.1007/s13205-018-1359-z -
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 -
Microbial Cell Factories Nov 2017Monascus pigments are widely used in the food and pharmaceutical industries due to their safety to human health. Our previous study found that glucose concentration...
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, HO 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 HO and DTT. Both cell growth and extracellular water-soluble yellow pigment production were enhanced under HO-induced oxidative (HIO) conditions and were inhibited under dithiothreitol-induced reductive conditions. By optimizing the amount of HO 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 HO 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 HO 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.
Topics: Color; Humans; Monascus; Oxidation-Reduction; Water
PubMed: 29162105
DOI: 10.1186/s12934-017-0828-0 -
Brazilian Journal of Microbiology :... 2018An ascomycetes fungus was isolated from brine storage of green olives of the Arauco cultivar imported from Argentina and identified as Monascus ruber. The combined...
An ascomycetes fungus was isolated from brine storage of green olives of the Arauco cultivar imported from Argentina and identified as Monascus ruber. The combined effects of different concentrations of sodium chloride (3.5-5.5%), sodium benzoate (0-0.1%), potassium sorbate (0-0.05%) and temperature (30-40°C) were investigated on the growth of M. ruber in the brine of stored table olives using a response surface methodology. A full 2 factorial design with three central points was first used in order to screen for the important factors (significant and marginally significant factors) and then a Face-Centered Central Composite Design was applied. Both preservatives prevented fungal spoilage, but potassium sorbate was the most efficient to control the fungi growth. The combined use of these preservatives did not show a synergistic effect. The results showed that the use of these salts may not be sufficient to prevent fungal spoilage and the greatest fungal growth was recorded at 30°C.
Topics: Food Preservation; Food Preservatives; Food Storage; Fruit; Hydrogen-Ion Concentration; Monascus; Olea; Sodium Benzoate; Sodium Chloride
PubMed: 29100931
DOI: 10.1016/j.bjm.2017.05.009 -
Experimental and Therapeutic Medicine Oct 2017Robusta beans cultivated with (RMR) were successively fermented with (LM) and the antiobesity effects were examined. To produce an obese mouse model to investigate the...
Robusta beans cultivated with (RMR) were successively fermented with (LM) and the antiobesity effects were examined. To produce an obese mouse model to investigate the hypolipidemic effects, ICR mice were fed the same high-fat diet for 6 weeks. Treatment groups were given 10 or 20% RMR-LM. Body weight changes in the 20% RMR-LM group were lower compared with those in the control group. Visceral adipose tissue weight and adipose size were significantly lower in the 20% RMR-LM group compared with those in the control group. Significant improvement in glucose tolerance was observed in the 10 and 20% RMR-LM groups compared with the control group. The 20% RMR-LM group exhibited a significant reduction in serum glucose concentration. Hepatic mRNA levels of sterol regulatory element-binding protein 1, fas cell surface death receptor, and peroxisome proliferator-activated receptor γ, which are associated with lipid, and fatty acid metabolism, in the 20% RMR-LM group were significantly lower compared with those in the control group. The results of the present study demonstrated that 20% RMR-LM may be used to prevent obesity, and ameliorate diabetes and lipid metabolism imbalances.
PubMed: 29042976
DOI: 10.3892/etm.2017.4990 -
Chemical Science Jul 2017azaphilone pigments (MonAzPs) are very widely used as food colorants, but their biosynthetic pathway has remained poorly characterized for more than half a century. In...
azaphilone pigments (MonAzPs) are very widely used as food colorants, but their biosynthetic pathway has remained poorly characterized for more than half a century. In this study, the individual steps of MonAzPs biosynthesis in M7 were elucidated by a combination of targeted gene knockouts, heterologous gene expression, and chemical and enzymatic reactions. This study describes the first rational engineering of MonAzPs biosynthesis and provides a roadmap for future pathway engineering efforts directed towards the selective production of the most valuable pigments and serves as a model for the biosynthesis of fungal azaphilones in general.
PubMed: 28959415
DOI: 10.1039/c7sc00475c -
AMB Express Aug 2017Monascus species are edible fungi due to the production of food colorant and other beneficial compounds. Hence, it has been an attractive thesis to improve their...
Monascus species are edible fungi due to the production of food colorant and other beneficial compounds. Hence, it has been an attractive thesis to improve their productivities. Increasing numbers of investigations revealed that regulating the activities of histone deacetylases can significantly perturb secondary metabolites (SM) production at a global level. In this study, dihydrocoumarin (DHC, an inhibitor of the Sirtuin family of NAD-dependent deacetylases) was used to treat Monascus ruber for evaluating its effects on organism growth and SM production. The results revealed that the variation trends of colonial sizes, biomass and mycotoxin were in a dose-dependent manner. Generally, they decreased with the increased DHC concentrations in the designed range. But the variation trend of pigment was different. Comparison of SM profile, three new peaks occurred to the mycelia extractions from DHC-treated strain corresponding to molecular weights 402, 416 and 444, respectively. These three compounds were identified as Monasfluol B, Monascus azaphilone C and acetyl-monasfluol B (a new Monascus chemical pigment structure). In short, DHC can stimulate M. ruber strain to produce more pigment-like polyketides but inhibition of mycotoxin (citrinin).
PubMed: 28836182
DOI: 10.1186/s13568-017-0467-1