GERI-BP001 Compounds, New Inhibitors of Acyl-CoA: Cholesterol Acyltransferase from Aspergillus fumigatus F37. I. Production, Isolation, and Physico-chemical and Biological Properties (original) (raw)
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The Journal of Antibiotics, 1996
A new inhibitor of acyl-CoA : cholesterol acyltransferase (ACAT), designated GERI-BP002-A, was isolated from the culture broth of Aspergillus fumigatus F93 by acetone extraction, EtOAc extraction, SiO2 column chromatography and reverse phase HPLC.Spectroscopic analyses of the compound identified bis (2-hydroxy-3-/m-butyl-5-methylphenyl) methane as the structure and its molecular weight and formula to be 340 and C23H32O2, respectively. GERI-BP002-A inhibited ACATactivity by 50%at the concentration of 50fiM in an enzyme assay system using rat liver microsomes.
2013
During the research for bioactive secondary metabolites from microorganisms, the endophytic fungi Aspergillus fumigatus sp. isolate R7 was found to produce a set of promising bioactive compounds (1-10) after its large scale fermentation, working up and purification using a series of chromatographic techniques. Structural elucidation of the yielded compounds using intensive studies of their NMR ( 1 H, 13 C& 2D NMR) and mass (EI MS, ESI MS) spectrometry confirmed them as linoleic acid (1), R(-)-glycerol monolinoleate (2), bis-dethio-(bis-methyl-thio)-gliotoxin (3), fumiquinazoline-F (4), fumiquinazoline-D (5), (Z,Z)-N,N’-[1-[(4-Hydroxy-phenyl)methylene]-2-[(4-methoxy-phenyl)-methylene]-1,2-ethanediyl]-bis-formamide (6), pyrazoline-3-one trimer (7), Tricho-9-ene-2a,3a,11a,16-tetraol (8), 2’-deoxy-thymidine (9), and cerebroside A (10). In this article, taxonomical characterization, fermentation, structural characterization of the obtained metabolites were reported together with their an...
Secondary metabolites and bioactivity of two fungal strains
Purpose The investigation of two fungal strains isolated from Egyptian habitats, namely, the endophytic Fusarium poae FUN1 and the terrestrial Penicillium italicum FUN2 to illustrate their chemical constituents and their bioactivities. Materials and methods See General instrumental procedures. Results Linoleic acid (1), indole-3-acetic acid methyl ester (2) and Nb-acetyltryptamine (3) were produced by F. poae FUN1, whereas P. italicum FUN2 also delivered linoleic acid (1) in addition to cis-cyclo-(prolyl,valyl) (4). The structures of compounds (1)–(4) were elucidated by 1D and 2D NMR, MS data and through comparison with literature reports. In this article, the taxonomical characterization of both fungal strains, their upscale fermentation and the antimicrobial and cytotoxic activities tested have been described. Conclusion Two different fungal strains, endophytic F. poae FUN1 and terrestrial P. italicum FUN2, were intensively studied biologically and chemically. Four bioactive compounds (1)–(4) were isolated, and structurally confirmed by intensive studies of NMR and MS. The antimicrobial and cytotoxic activities of the fungal extracts and their delivered compounds were studied. This might be helpful for the cure of recent diseases, and drug-resistant phenomena as well as in the development of pharmaceutical, agrochemical and biochemical agents and their lead compounds.
Chemical investigation of novel ascomycetes using PCR based screening approaches
World Journal of Microbiology & Biotechnology
Fungi are well known for a wealth of pharmacologically important activities and agrochemical properties. Polyketides that are widely found in fungi, are a large group of secondary metabolites which exhibit diversity in their function and structure. Here we described an investigation of three fungal strains which were prospected for production of polyketides. The aim of this work was to employ the diversity of reducing type I polyketide synthase genes in these fungi using a molecular and bioinformatics approaches as a mini tool. A degenerate primer pair for highly reduced PKSs was newly designed and used together with ketosynthase primers for amplification. One hundred and thirty-eight clones were sequenced. Ten KS domain sequences were isolated, using two primer pairs specific for highly reduced type PKSs. This study revealed four sequences from Emarcea castanopsidicola, four ketosynthase sequences from Gaeumannomyces amomi and two sequences from Leiosphaerella amomi, respectively. Bioinformatic techniques were employed to identify a group of these KS domain sequences. Based on these sequences suggested that rapid screening provided the potential to explore significant PKS structural diversity. Hence chemical investigation had been conducted and exhibited nine compounds. The endophytic fungus L. amomi was cultivated and elucidated linoleic acid, ergosterol and an unidentified sterol in the extracts. Linoleic acid, sitosterol, and p-hydroxybenzoic acid were isolated from the saprobic fungus E. castanopsidicola. We first isolated a new polyketide, stemphol 1-O-β-D-galactopyranoside together with four known metabolites; stemphol, kojic acid, ergosterol, indole-3-carboxylic acid from an ethyl acetate extract of the cultures of G. amomi. Stemphol was classified as a phenolic lipid or resorcinolic lipid, which have biopharmacological, biomedical, and biotechnological importance. However, recent researches have revealed that these molecule types are synthesized by 2′-oxoalkylresorcinolic acid synthase. The prospective KS domain sequences from this study will be used as probes to isolate putative PKS genes. A gene cluster responsible for PK biosynthesis should be confirmed by determination of PK products generated by these enzymes.
Induction of diverse secondary metabolites in Aspergillus fumigatus by microbial co-culture
RSC Advances, 2013
An established culture of Aspergillus fumigatus MBC-F1-10 proved to be very receptive to external stimuli and reacted with the production of secondary metabolites which were undetectable when the fungus was grown under standard conditions. Firstly, co-cultivation with the type strain of Streptomyces bullii, an isolate from hyper-arid Atacama desert soil, led to the isolation of ergosterol 1, seven metabolites belonging to the diketopiperazine alkaloids; brevianamide F 2, spirotryprostatin A 3, 6-methoxy spirotryprostatin B 4, fumitremorgin C and its 12,13-dihydroxy derivative (5-6), fumitremorgin B 7, and verruculogen 8, in addition to 11-O-methylpseurotin A 9 and its new isomer 11-O-methylpseurotin A 2 10. In an independent experiment, addition of N-butyryl-DL-homoserine lactone to the culture medium led to the production of emestrins A and B (11-12). Neither microbe produced these compounds when cultured alone. The structures of all compounds were elucidated using NMR spectroscopic techniques and mass spectrometric analysis. The isolated compounds were tested for their potential antibacterial and antiprotozoal activities.
Isolation and characterization of fungal strains as biocontrol agents
Agriculture is the back-bone of country and Indian economy (K Kabir). Plant pathogenic bacteria and fungi are considered economically important around the world (Carson 1962; Houeto et al. 1995) due to cause many serious diseases of plants (Vidhyasekaran 2002). Modern agronomy, pesticides and fertilizers have sharply increased yields from cultivation, but also caused widespread ecological damage and negative human health effects (EPA 2006). So, there is a need to replace conventional pesticides and fertilizers. Microorganisms with ability to suppress disease causing fungi, bacteria and insect-pests are potentially important alternatives to chemical pesticides and have been reported by many researchers. With the above facts the present study entitled “Isolation and Characterization of Fungal Strains as Biocontrol Agents.” In the study Fungal strains namely Trichoderma viride, Acrimonium strictum, Aspergillus terreus, Aspergillus oryzae, Aspergillus niger, Paecilomyces variotii, Aspergillus fumigates and Penicillium glabrum were found antibacterial active against 11 plant pathogenic bacterial strains. These fungi having antimicrobial activity can be used as biopesticides against broad range of pathogenic micro-organisms like bacteria & fungi. So the study will be helpful to improve crop yield as well as quality.
Brazilian Journal of Microbiology, 2021
The main objective of the study is to characterize two new strains of Aspergillus fumigatus through morphometric, biochemical, molecular methods, and to evaluate their antimicrobial potentiality. The micro-morphotaxonomy, growth, and metabolic behavior of the strains, nHF-01 and PPR-01, were studied in different growth conditions and compared with standard strain. The molecular characterization was done by sequencing the ncrDNA ITS1-5.8S-ITS2 and D1–D2 domains of the nc 28S rDNA region and compared with a secondary structure-based phylogenetic tree. The secretory antimicrobials and pigments were characterized by TLC, UV-Vis, and FT-IR spectroscopy. Both the strains showed distinct growth patterns in different nutritional media and could assimilate a wide range of carbohydrates with distinctive biochemical properties. The molecular characterization revealed the strains, nHF-01 and PPR-01, as Aspergillus fumigatus (GenBank Accession No. {"type":"entrez-nucleotide","attrs":{"text":"MN190286","term_id":"1706844409"}}MN190286 and {"type":"entrez-nucleotide","attrs":{"text":"MN190284","term_id":"1706844407"}}MN190284, respectively). It was observed that the strain nHF-01 produces red to brownish pigments having mild antimicrobial activity while the strain PPR-01 does not represent such transformations. The extractable compounds had a significant antimicrobial potentiality against the human pathogenic bacteria. From this analysis, it can be concluded that the nHF-01 and PPR-01 strains are distinct from other A. fumigatus by their unique characters. Large-scale production and detailed molecular elucidation of the antimicrobial compounds may lead to the discovery of new antimicrobial compounds from these strains.Supplementary InformationThe online version contains supplementary material available at 10.1007/s42770-021-00439-w.
Iranian Biomedical Journal
Background: The majority of studies on soil Aspergillus concern the isolation and characterization of the antimicrobial compounds produced by this organism. Our previous studies indicated an isolated Aspergillus strain soil to be of interest, and this subject is further investigated here. Method: Soil samples of various locations in Iran were collected. Extract from Aspergillus sp. culture was obtained using ethyl acetate fractionation. Antimicrobial activity testing was performed using broth microdilution assay against Escherichia coli, Candida albicans, and Staphylococcus aureus microorganisms. One metabolite PA3-d10 was isolated from these active extracts and identified using thin layer chromatography, preparative thin-layer chromatography, HPLC, 1 HNMR (proton nuclear magnetic resonance), 2D NMR, and LC-MS (liquid chromatography-mass spectrometry). Results: According to morphological and biochemical properties as well as ITS rDNA sequencing, we identified an isolate of Aspergillus flavus. The ethyl acetate fraction of the fermentation medium containing membrane active metabolites showed antimicrobial effects against different bacterial and yeast indicator strains. One metabolite from these active extracts was finally identified. Conclusion: Membrane active fraction produced by Aspergillus strain in this research demonstrated antimicrobial activities against bacteria and yeast strains. Therefore, this metabolite can be considered as a potential antimicrobial membrane active agent.
Characterization of kefir yeasts with antifungal capacity against Aspergillus species
Research Square (Research Square), 2022
Ke r is a fermented probiotic drink obtained by placing ke r granules in a suitable substrate. The ke r granules are a consortium of bacteria and yeasts embedded in a exopolysaccharide matrix. The aim of this research was the isolation and identi cation of yeasts from ke r of different origin, the evaluation of their antifungal capacity against Aspergillus spp. and the characterization of virulence related traits. Using RFLP of ITS1/ITS4 region, D1/D2 region sequencing and RAPD techniques, 20 ke r isolates were identi ed as Geotrichum candidum, Pichia kudriavzevii, P. membranifaciens, Saccharomyces cerevisiae and Candida ethanolica. Their antifungal capacity was evaluated by their conidia germination reduction, which allowed the selection of eight isolates with high to moderate conidia germination reduction against A. avus and A. parasiticus. Furthermore, these selected isolates showed growth inhibition on contact in the dual culture assay for both Aspergillus species and 3 of them-belonging to S. cerevisiae and P. kudriavzevii species-generated volatile organic compounds which signi cantly affected the growth of both fungi. For the evaluation of virulence related traits, growth at high temperatures, enzymatic activities and the adhesion to Caco-2 cells were analyzed. The isolates did not present more than one positive virulence-related trait simultaneously. In particular, it is important to highlight that the adhesion capacity to the model of intestinal barrier was extremely low for all of them. According to the results obtained, further studies would be of interest for the possible use of these promising yeasts as biocontrol agents against fungi in food. Water ke r granules-CMUNLP 1, CMUNLP 2 and CMUNLP 4-and milk ke r granules-CIDCA AGK1, CMUNLP 8 and CMUNLP 9-employed in this work, were from different households of La Plata, Argentina, with the exception of CMUNLP1 which was from India and AGK1 which belongs to the collection of Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA, UNLP, Argentina). The ke r granules were cultivated in bottles of 250 ml capacity with different substratesmilk (M) or whey permeate (WP) for milk ke r and muscovado (MU), molasses (MO) or chancaca (CC) for water ke r-in a granule/substrate ratio of 10% for 48 h at 30°C. Subsequently, the granules were separated from de supernatant by sieving and the granules were incorporated into the respective fresh substrate. This process was repeated three consecutive times before yeast isolation from granules and supernatant (Gamba et al., 2016a). Yeast isolation The yeasts isolation was carried out from the granules and supernatants obtained in the previous step, for which 10 g and 10 ml respectively, were added 90 ml of 0.85% w/v NaCl and homogenized in a BagMixer® 400 W (Interscience, France). Serial dilutions were made and 100 µl of the nal dilutions were surface-spread on Petri dishes containing yeast glucose chloramphenicol agar (YGC, Biokar, Francia), in duplicate for each dilution. The incubation was carried out for 5 d at 30°C and the colonies obtained were differentiated according to their morphological characteristics, then subcultivated on Petri dishes with yeast peptone dextrose agar (YPD, 1% yeast extract, 2% peptone, 2% dextrose, 2% agar) and nally maintained in YPD slants at 4°C. Yeast identi cation Genomic DNA was extracted according to Lõoke et al. (2011). Firstly, a restriction fragment length polymorphism (RFLP) of the ITS1/ITS2 region was performed according to Esteve-Zarzoso et al. (1999). From the results obtained the yeasts were identi ed and grouped, then the results were con rmed by amplifying and sequencing the D1/D2 region according to (Kurtzman and Robnett, 1997). The PCR products were puri ed with MinElute® according to the manufacturer´s instructions and their sequencing was performed at the Genomic Unit-Valencia University (Valencia, Spain) and Macrogen (Seoul, South Korea). The identi cation was achieved by the comparison against the Genbank database using BLAST (http://blast.ncbi.nlm.nih.gov/Blast.cgi). Additionally, for the identi cation of Geotrichum candidum a randomly ampli ed polymorphic DNA (RAPD) analysis with the primer M13 (5'-GAGGGTGGCGGTTCT-3') was performed (Gente et al., 2006). In the case of Saccharomyces cerevisiae, an additional RFLP of the genes MAG2 and GSY1 was performed according to Pérez-Tráves et al. (2014). Preparation of cell free supernatants (CFS) The identi ed yeasts were cultured in 50 ml of MEA broth (10 g/l malt extract, 20 g/l yeast extract) in 250 ml asks, with two previous passages in the same medium, and incubated at 30°C for 48 h at 150 rpm. Afterwards, the fermented medium was centrifugated at 3000 rpm for 15 min and the supernatant was sterilized by ltration using 0.22 µm pore acetate membranes (Sigma-Aldrich®). MEA dishes streaked
Comparative Chemistry of Aspergillus oryzae (RIB40) and A. flavus (NRRL 3357)
Metabolites, 2012
Aspergillus oryzae and A. flavus are important species in industrial biotechnology and food safety and have been some of the first aspergilli to be fully genome sequenced. Bioinformatic analysis has revealed 99.5% gene homology between the two species pointing towards a large coherence in the secondary metabolite production. In this study we report on the first comparison of secondary metabolite production between the full genome sequenced strains of A. oryzae (RIB40) and A. flavus (NRRL 3357). Surprisingly, the overall chemical profiles of the two strains were mostly very different across 15 growth conditions. Contrary to previous studies we found the aflatrem precursor 13-desoxypaxilline to be a major metabolite from A. oryzae under certain growth conditions. For the first time, we additionally report A. oryzae to produce parasiticolide A and two new analogues hereof, along with four new alkaloids related to the A. flavus metabolites ditryptophenalines and miyakamides. Generally the secondary metabolite capability of A. oryzae presents several novel end products likely to result from the domestication process from A. flavus.