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Microorganisms Jan 2020species have been used for the production of many industrially and medically important metabolites, most of which are polyketides produced by the action of polyketide...
species have been used for the production of many industrially and medically important metabolites, most of which are polyketides produced by the action of polyketide synthases that use acetyl-CoA and malonyl-CoA as precursors, and some of them are derived from acetate. In this study the effects of acetic acid, and two kinds of acetates, sodium acetate and ammonium acetate at different concentrations (0.1%, 0.25% and 0.5%) on the morphologies, biomasses, and six major pigments (MPs) of M7 were investigated when M7 strain was cultured on potato dextrose agar (PDA) at 28 °C for 4, 8, 12 days. The results showed that all of the added acetate species significantly affected eight above-mentioned parameters. In regard to morphologies, generally the colonies transformed from a big orange fleecy ones to a small compact reddish ones, or a tightly-packed orange ones without dispersed mycelia with the increase of additives concentration. About the biomass, addition of ammonium acetate at 0.1% increased the biomass of M. ruber M7. With respect to six MPs, all acetate species can enhance pigment production, and ammonium acetate has the most significant impacts. Production of monascin and ankaflavin had the highest increase of 11.7-fold and 14.2-fold in extracellular contents at the 8th day when 0.1% ammonium acetate was supplemented into PDA. Intracellular rubropunctatin and monascorubrin contents gained 9.6 and 6.46-fold at the 8th day, when 0.1% ammonium acetate was added into PDA. And the extracellular contents of rubropunctamine and monascorubramine were raised by 1865 and 4100-fold at the 4th day when M7 grew on PDA with 0.5% ammonium acetate.
PubMed: 31936171
DOI: 10.3390/microorganisms8010081 -
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) Mar 2021The effects of a static magnetic field (SMF) on M7 ( M7) cultured on potato dextrose agar (PDA) plates under SMF treatment at different intensities (5, 10, and 30 mT)...
The effects of a static magnetic field (SMF) on M7 ( M7) cultured on potato dextrose agar (PDA) plates under SMF treatment at different intensities (5, 10, and 30 mT) were investigated in this paper. The results revealed that, compared with the control (CK, no SMF treatment), the SMF at all tested intensities did not significantly influence the morphological characteristics of M7, while the intracellular and extracellular pigments (MPs) and extracellular citrinin (CIT) of M7 were increased at 10 and 30 mT SMF but there was no impact on the MPs and CIT at 5 mT SMF. The transcriptome data of M7 cultured at 30 mT SMF on PDA for 3 and 7 d showed that the SMF could increase the transcriptional levels of some relative genes with the primary metabolism, including the carbohydrate metabolism, amino acid metabolism, and lipid metabolism, especially in the early growing period (3 d). SMF could also affect the transcriptional levels of the related genes to the biosynthetic pathways of MPs, CIT, and ergosterol, and improve the transcription of the relative genes in the mitogen-activated protein kinase (MAPK) signaling pathway of M7. These findings provide insights into a comprehensive understanding of the effects of SMF on filamentous fungi.
PubMed: 33808107
DOI: 10.3390/jof7040256 -
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 -
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 -
3 Biotech Sep 2022In this study, the whole genome of KACC 46666 was generated using the PacBio RSII sequencer with high-quality de novo assembly to obtain trustworthy assembly and...
In this study, the whole genome of KACC 46666 was generated using the PacBio RSII sequencer with high-quality de novo assembly to obtain trustworthy assembly and annotation using genome assemblies with long reads from PacBio single-molecule real-time sequencing. The whole genome of has a total length of 25.9 Mb, divided in 13 contigs with 9639 genes. The functions of genes involved in secondary metabolite production were further analyzed. Gene clusters involved in the production of pigment, monacolin K, and mycotoxin citrinin were identified. Notably, most of the citrinin gene cluster was lost, as confirmed via high-performance liquid chromatography analysis. This genome-level safety evaluation of industrially important strains will provide valuable information for genome-based microbial engineering of natural food colorants and production of commercially important secondary metabolites such as monacolin K.
PubMed: 35959167
DOI: 10.1007/s13205-022-03287-z -
Mycotoxin Research May 2022Fungi and mycotoxins in silage can have detrimental consequences for both cattle and human health. This pilot study identified, via the routinary direct plating method,...
Fungi and mycotoxins in silage can have detrimental consequences for both cattle and human health. This pilot study identified, via the routinary direct plating method, the dominant cultivable fungi in mouldy grass silages (GS) (n = 19) and maize silages (MS) (n = 28) from Austria. The profiles of regulated, modified, and emerging mycotoxins together with other fungal metabolites were analysed via LC-(ESI)MS/MS. Penicillium roqueforti, Saccharomyces spp., Geotrichum candidum, Aspergillus fumigatus and Monascus ruber were the most frequent fungal organisms identified. Other species including Mucor circinelloides, Fusarium spp. and Paecilomyces niveus were detected at lower frequencies. The presence of complex mixtures of toxic and potentially toxic compounds was evidenced by high levels and occurrences (≥ 50%) of Penicillium-produced compounds such as mycophenolic acid (MPA), roquefortines (ROCs), andrastins (ANDs) and marcfortine A. Mouldy silages contained toxins commonly produced by genus Fusarium (e.g. zearalenone (ZEN) and trichothecenes), Alternaria (like tenuazonic acid (TeA) and alternariol (AHO)) and Aspergillus (such as sterigmatocystin (STC)). Compared to those in GS, mouldy spots in MS presented significantly higher fungal counts and more diverse toxin profiles, in addition to superior levels of Fusarium spp., Penicillium spp. and total fungal metabolites. Generally, no correlation between mould counts and corresponding metabolites was detected, except for the counts of P. roqueforti, which were positively correlated with Penicillium spp. metabolites in mouldy MS. This study represents a first assessment of the fungal diversity in mouldy silage in Austria and highlights its potential role as a substantial contributor to contamination with complex mycotoxin mixtures in cattle diets.
Topics: Alternaria; Animals; Austria; Cattle; Food Contamination; Fusarium; Mycotoxins; Pilot Projects; Poaceae; Silage; Tandem Mass Spectrometry; Zea mays
PubMed: 35347677
DOI: 10.1007/s12550-022-00453-3 -
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 -
Journal of Fungi (Basel, Switzerland) Dec 2023Citrinin (CIT), a secondary metabolite produced by the filamentous fungi species, exhibits nephrotoxic, hepatotoxic, and carcinogenic effects in mammals, remarkably...
Citrinin (CIT), a secondary metabolite produced by the filamentous fungi species, exhibits nephrotoxic, hepatotoxic, and carcinogenic effects in mammals, remarkably restricting the utilization of -derived products. CIT synthesis is mediated through the gene and modified by multiple genetic factors. Here, the regulatory effects of two transcripts, α, and β, generated via pre-mRNA alternative splicing (AS), were investigated using hairpin RNA (ihpRNA) interference, and their impact on CIT biosynthesis and the underlying mechanisms were assessed through chemical biology and transcriptome analyses. The CIT yield in ihpRNA-pksCTα and ihpRNA-pksCT (α + β) transformants decreased from 7.2 μg/mL in the wild-type strain to 3.8 μg/mL and 0.08 μg/mL, respectively. Notably, several genes in the CIT biosynthetic gene cluster, specifically , , and in the ihpRNA-pksCT (α + β) transformant, were downregulated. Transcriptome results revealed that silencing has a great impact on carbohydrate metabolism, amino acid metabolism, lipid metabolism, and AS events. The key enzymes in the citrate cycle (TCA cycle) and glycolysis were significantly inhibited in the transformants, leading to a decrease in the production of biosynthetic precursors, such as acetyl-coenzyme-A (acetyl-coA) and malonyl-coenzyme-A (malonyl-coA). Furthermore, the reduction of CIT has a regulatory effect on lipid metabolism via redirecting acetyl-coA from CIT biosynthesis towards lipid biosynthesis. These findings offer insights into the mechanisms underlying CIT biosynthesis and AS in , thus providing a foundation for future research.
PubMed: 38132775
DOI: 10.3390/jof9121174