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Surface growth of fungi in 1-ml static fermentations. (a) Synnemata of Rhizostilbella hibisci. (b) Exudates and mycelial mat of an unidentified coelomycete. (c) Ascomata of Ophiostoma sp. Inner diameter of well = 8 mm.
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We asked to what extent does the application of the OSMAC (one strain, many compounds) approach lead to enhanced detection of antibiotics and secondary metabolites in fungi? Protocols for bacterial microfermentations were adapted to grow fungi in nutritional arrays.
Protocols for microfermentations of non-sporulating fungi were validated using know...
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Context 1
... exhibited elaborate morphological differentiation comparable with that of agar cultures or solid state fermentations on seed-based or vermiculite-based media. Differentiation was manifested in natural and sometimes intense pigmentation, pigmented exudations and production of complex reproductive structures, e.g. stromata, pycnidia, and ascomata (Fig. 3). Cross contamination of strains between adjacent wells occurred rarely, if at all. In nearly all cases when contamination occurred, its source was from the inoculum tubes. Usually, such contamination could be detected by replicating the master plate onto agar media that promoted fungal or bacterial growth (e.g. Fig. ...
Context 2
... Morinia pestalozzioides (Fig. 6a, column 2), Glarea lozoyensis (Fig. 6a, column 8) and Emericella rugulosa (Fig. 6a, column 12) produced antifungal metabolites on most of the media set. Extracts of Monascus purpureus (Fig. 6a, column 10) were only weakly active, and hazy ZOIs were evident on most media. Other strains, e.g. Arthrinium arundinis (Fig. 6b, column 3), Monocillium sp. (Fig. 6a, column 4), Xylaria sp., (Fig. 6b, column 5), Leiothecium sp. (Fig. 6a, column 6), Paecilomyces lilacinus (Fig. 6b, column 7), Favolaschia pustulata (Fig. 6b, column 9) and Paecilomyces inflatus (Fig. 6b, column 11) showed medium-specific production of antifungal activity. When the same set was replicated onto ...
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Citations
... The culture condition variation alters the expression of the biosynthestic gene expression responsible for structural diversity and production of the metabolites [36]. Therefore, the isolated fungi were incubated at 25 • C for seven days on the agar medium such as Czapek-Dox (CZ), Potato-Dextrose (PD), Malt Extract (ME), and Yeast Extract Sucrose (YES), to identify the fungal-media combinations having the ability to metabolically utilize various substrates such as carbohydrates, and compounds as sole sources of carbon. ...
Some fungi have a unique ability to adapt and establish beneficial association with crop in salt–affected soils. Therefore, present study was conducted to characterize the salinity–tolerance, P–solubilization and growth promotion ability of the fungal isolates from the rhizosphere of salt–tolerant crops. The rhizospheric soil showed an assemblage of fifteen phosphate–solubilizing fungal (PSF) isolates tolerant to salinity (5% NaCl w/v), alkalinity (pH 8.0), and thermal stress (up to 40oC). Soil pH, EC, organic carbon, KMnO4–N, Olsen’s P, and NH4OAc–K explained about 51.6% variability in the rhizosphere assemblage of the microorganisms. The genera Aspergillus and Penicillium were abundant in rhizosphere. The association of Penicillium spp. with crops was greater in soil of higher salinity than Aspergillus. The salt–tolerant fungi demonstrated variable effects on the germination of different crops. Siderophore and fungal biomass of the isolates cause 23% variability in plant growth parameters. The PSF strains PP3 and SA1 of the Penicillium and Aspergillus, respectively, showed high P solubilization without any appreciable change at high salinity. This study concluded that the PSF producing siderophores and phytohormones has significant importance for promoting growth and survival of crops in salt–affected soils.
... Bioactive secondary metabolites are a broad class of organic compounds with unique chemical structures and functions that are produced in large quantities by fungi [1]. These secondary metabolites, also known as natural products [1], are vital to the survival and adaptation of fungi in their ecological interactions [2]. A lot of attention is focused on these bioactive secondary metabolites' possible anticandida properties, or their capacity to fight infections brought on by Candida species [3]. ...
... In the field of fungal discoveries, penicillin produced by Penicillium notatum has been considered a goldmine since 1928 [35]. Ascomycetes such as Aspergillus, Penicillium, and Fuserium, as well as imperfect fungi, are common producers of bioactive compounds, followed by the endophytic [35][36][37] and filamentous species, and Alterneria, Trichoderma, and Phoma [38]. However, there are still several metabolites of these microorganisms that are yet to be characterized [39]. ...
Fungi are the most diverse living organisms on planet Earth, where their ubiquitous presence in various ecosystems offers vast potential for the research and discovery of new, naturally occurring medicinal products. Concerning human health, cancer remains one of the leading causes of mortality. While extensive research is being conducted on treatments and their efficacy in various stages of cancer, finding cytotoxic drugs that target tumor cells with no/less toxicity toward normal tissue is a significant challenge. In addition, traditional cancer treatments continue to suffer from chemical resistance. Fortunately, the cytotoxic properties of several natural products derived from various microorganisms, including fungi, are now well-established. The current review aims to extract and consolidate the findings of various scientific studies that identified fungi-derived bioactive metabolites with antitumor (anticancer) properties. The antitumor secondary metabolites identified from extremophilic and extremotolerant fungi are grouped according to their biological activity and type. It became evident that the significance of these compounds, with their medicinal properties and their potential application in cancer treatment, is tremendous. Furthermore, the utilization of omics tools, analysis, and genome mining technology to identify the novel metabolites for targeted treatments is discussed. Through this review, we tried to accentuate the invaluable importance of fungi grown in extreme environments and the necessity of innovative research in discovering naturally occurring bioactive compounds for the development of novel cancer treatments.
... Therefore, altering the nutrition and the surrounding environment by changing the medium composition, aeration, pH, and culture vessel may provide optimal conditions to produce a wide variety of metabolites. This technique is called OSMAC (one strain, many compounds) (Bills et al. 2008). Bacterial growth favors the neutral or slightly alkaline condition, while slight pH change can significantly affect the production of secondary metabolites (Miao et al. 2006). ...
Sponges are habitats for a diverse community of microorganisms. Sponges provide shelter, whereas microbes provide a complementary defensive mechanism. Here, a symbiotic bacterium, identified as Bacillus spp., was isolated from a marine sponge following culture enrichment. Fermentation-assisted metabolomics using thin-layer chromatography (TLC) and gas chromatography-mass spectrometry (GC-MS) indicated that marine simulated nutrition and temperature was the optimum in metabolite production represented by the highest number of metabolites and the diverse chemical classes when compared to other culture media. Following large-scale culture in potato dextrose broth (PDB) and dereplication, compound M1 was isolated and identified as octadecyl-1-(2′,6′-di-tert-butyl-1′-hydroxyphenyl) propionate. M1, at screening concentrations up to 10 mg/ml, showed no activity against prokaryotic bacteria including Staphylococcus aureus and Escherichia coli, while 1 mg/ml of M1 was sufficient to cause a significant killing effect on eukaryotic cells including Candida albicans, Candida auris, and Rhizopus delemar fungi and different mammalian cells. M1 exhibited MIC50 0.97 ± 0.006 and 7.667 ± 0.079 mg/ml against C. albicans and C. auris, respectively. Like fatty acid esters, we hypothesize that M1 is stored in a less harmful form and upon pathogenic attack is hydrolyzed to a more active form as a defensive metabolite. Subsequently, [3-(3,5-di-tert-butyl-4-hydroxyphenyl)-propionic acid] (DTBPA), the hydrolysis product of M1, exhibited ~ 8-fold and 18-fold more antifungal activity than M1 against C. albicans and C. auris, respectively. These findings indicated the selectivity of that compound as a defensive metabolite towards the eukaryotic cells particularly the fungi, a major infectious agent to sponges. Metabolomic-assisted fermentation can provide a significant understanding of a triple marine-evolved interaction.
Key points
• Bacillus species, closely related to uncultured Bacillus, is isolated from Gulf marine sponge
• Metabolomic-assisted fermentations showed diverse metabolites
• An ester with a killing effect against eukaryotes but not prokaryotes is isolated
... It is of great interest in the study of the physiology of a microorganism to test the use of different culture media, since it has been observed that the complexity of the medium has a significant influence on the growth of the mycelium and, therefore, on the speed of biomass production [11,12]. To achieve a higher production of these antifungal or bioactive metabolites, optimal conditions must be ensured during the trophophase for good development of the microorganism [13,14]. ...
In this study, the characteristic growth of Cladobotryum virescens on nine culture media was analyzed. The growing behavior of this fungus was dependent on the culture medium. In vitro analysis showed that oat agar was better than other media tested with the highest conidia production. The antifungal activity against Fusarium chlamydosporum and Alternaria brassicicola was evaluated by the Dual Culture method. C. virescens displayed high activity against both pathogens acting through antibiosis and mycoparasitism. This effect was increased by a higher competitiveness of the strain for the substrate. Furthermore, the crude ethyl acetate extract of the culture broth was tested in vitro against Botrytis cinerea and Septoria tritici, as well as the hemibiotrophic oomycete Phytophthora infestans using a microtiter plate assay at different concentrations. The extract showed excellent inhibition even below 5 ppm. According to these results, we concluded that C. virescens can be considered as a potential biological control agent in agriculture. To the best of our knowledge, this is the first study to investigate C. virescens as a biocontrol agent for different diseases caused by five relevant pathogens that affect cereals and vegetables.
... Fungi are the most ubiquitous organism in nature as they are capable to synthesize broad range of compounds naturally from simple molecules which referred to as primary metabolites to the complex molecules are called secondary metabolites (Rahi and Malik, 2016). Although the commercial production of fungal metabolites depends on the amount of nutrients, its growth environments and culture conditions that promotes metabolite biosynthesis (Bills et al., 2008, Moreira et al., 2003Barberel and Walker, 2000). Different chemicals or substrates are generally used to transform the actions of cellular enzymes by the organisms for the production of bioactive metabolites that might be used as building blocks for the production of secondary metabolites (Nielsen, 2018, Bezalel et al., 1996Agrawal et al., 2016). ...
White rot fungi is a well known wood decaying macro-fungi being studied for many years for the potential applications in the bio-degradation of environmental pollutants because they produces a varieties of enzyme primarily responsible for the degradation of complex organic polymers. Along with these white rot fungi produces a vast number of cellular and extracellular metabolites having significant biological properties like antimicrobial, antioxidant, immunomodulatory, and nutritional properties. Because of these properties white rot fungi are being used in many field of science like Environmental Biotechnology, Bio-remediation technology, dye decolourization technology, Pharmaceutical industries, Nutritional researches etc. Aim of the study is to find out the different types of metabolites they produce, in which bio-synthetic pathways and their significant biological properties.
... Compositions of the media and their manipulation show success in the discovering natural products. These compositions lead to fungal secondary metabolites, if any nutritional variations of the medium can affect the on a production of the metabolites (15,16). Related to the media, R. oryzae can increase produced lipase and pectinase when growing in a medium containing carbon source supplemented with olive oil. ...
Rhizopus oryzae is classified a one of the Mucorales which belong to the zygomycota. This microorganism is a filamentous fungus can produce various substances such as ethanol, fumaric acid, lactic acid, ragi, and temph. R. oryzae has been occupied important role in the producing benefit metabolites such as organic acids. Enzymes. and antimicrobial compounds. In the current study, potato dextrose agar (PDA) and malt extract agar (MEA) were used for growing R. oryzae that the fungus was identified depending on the morphological parameters and conventional PCR. The fungus was cultured in two synthetic media were corn steep liquor (SCL) and conocarpus leaves (CL) at 25°C. R. oryzae produced abundant growth in both media and the growth was greyish white. GC-MA analysis appeared several different compounds in the crude extracts of R. oryzae using these fermenting synthetic media; however, some compounds were reported to be similar in their structures. The similar compounds were (pyrrolo [1,2-a] pyrazine-1,4-dione, hexahydro-), (1-nonadecene), (pyrrolo [1,2-a] pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl),) methyl stearate), (hexanedioic acid, bis(2-ethylhexyl) ester), (bis(2-ethylhexyl) phthalate), and (1,4-benzenedicarboxylic acid, (bis (2-ethylhexyl) ester). This study aimed to evaluate CL medium in comparison with CSL one for detecting the natural compounds produced from R. oryzae.
... In addition to nutrient compositions in culture media affecting cellular productivity, the cultural media design is therefore crucial in producing secondary metabolites (Atalla et al., 2008). Fungi cultivation is engineered to achieve the production of specific or new compounds by isolating the fungus and cultivating it in modified media (Bills et al., 2008). Tweaking microbial culture parameters to increase the diversity of metabolism production is referred to the One Strain, Many Compounds (OSMAC) approach. ...
... The quantity and diversity of bioactive compound production differ based on the media's nutrient compositions and culture conditions (Bills et al., 2008). The modified cultivation parameters such as medium composition affect chemical diversity and improve the yields of new microbial bioactive compounds produced by microorganisms (Romano et al., 2018;Gao et al., 2020). ...
Highlight Research Fusarium symbiont Agelas sponge has antibacterial activity against aureus and E. coli Multi Drug Resistant Fusarium can grow on modified media soursop juice (SJ) and ginger juice (GJ) and has different inhibitory abilities against bacteria test Fusarium bioactive compounds were analyzed and purified by HPLC Based on TLC visualization, one of the bioactive compounds contained in Fusarium is a steroid group Abstract Marine symbiont fungi, specifically, are likely to have antibacterial properties. The production of secondary metabolites in cultures is strongly influenced by nutrient availability. This study aimed to study the modified media for the cultivation of Fusarium sp. and its antibacterial bioactivity. Fusarium sp. was isolated from Agelas sp. sponge collected from Riung Sea, East South Nusa, Indonesia. In this study, the modified media was soursop juice (SJ), ginger juice (GJ), and sago (S). The fungal mycelium was cultured and scaled up for 7–14 days until the mycelium achieved maximum growth. A filtration funnel and HPLC were used to purify the bioactive compounds. The diffusion agar method was used to test antibacterial activity against the multi-drug resistant (MDR) Staphylococcus aureus and Escherichia coli. The results indicated that Fusarium sp. could grow on soursop juice and ginger juice but could not grow on sago media. The inhibition zone produced by the filtration fraction of Fusarium sp. from each media differed. The soursop juice media produced the largest inhibition zone against both S. aureus (11.56 mm ± 0.140) and E. coli (12.16 mm ± 0.094) at 100 μg/disc. The ginger juice (GJ) and soursop juice (SJ) media are promising as alternative culture media for Fusarium sp. The structure of the bioactive compound Fusarium sp. from GJ and MJ media culture can be further investigated using NMR.
... Variation of media type and composition falls within the framework of the one strain many-compounds (OSMAC) approach (Bode et al., 2002). This method entails systematic manipulation of various growth conditions with an aim of identifying the growth regimes that enhances optimum quality, diversity, and novelty of bioactive secondary metabolites (Bills et al., 2008;VanderMolen et al., 2013). Thus, variation of media type and composition can be employed to enhance the yield of a target bioactive compound (García-Kirchner et al., 2005) or as an approach to maximize the chemical diversity from an individual strain or few selected strains (Elias et al., 2006;VanderMolen et al., 2013). ...
Endophytic microorganisms are an important source of bioactive secondary metabolites. In this study, fungal endophytes obtained from A*STAR’s Natural Product Library (NPL) and previously isolated from different habitats of Singapore were investigated for their diversity, antimicrobial, and cytotoxic activities. A total of 222 fungal strains were identified on the basis of sequence analysis of ITS region of the rDNA gene. The identified fungal strains belong to 59 genera distributed in 20 orders. Majority of the identified strains (99%; 219 strains) belong to the phylum Ascomycota , while two strains belonged to the phylum Basidiomycota , and only one strain was from Mucoromycota phylum. The most dominant genus was Colletotrichum accounting for 27% of all the identified strains. Chemical elicitation using 5-azacytidine and suberoylanilide hydroxamic acid (SAHA) and variation of fermentation media resulted in the discovery of more bioactive strains. Bioassay-guided isolation and structure elucidation of active constituents from three prioritized fungal strains: Lophiotrema sp. F6932, Muyocopron laterale F5912, and Colletotrichum tropicicola F10154, led to the isolation of a known compound; palmarumycin C 8 and five novel compounds; palmarumycin CP 30 , muyocopronol A-C and tropicicolide. Tropicicolide displayed the strongest antifungal activity against Aspergillus fumigatus with an IC 50 value of 1.8 μg/ml but with a weaker activity against the Candida albicans presenting an IC 50 of 7.1 μg/ml. Palmarumycin C 8 revealed the best antiproliferative activity with IC 50 values of 1.1 and 2.1 μg/ml against MIA PaCa-2 and PANC-1 cells, respectively.
... Our screening pipeline involves growing fungi in 24 well plates on multiple growth media. It has previously been shown that different culture conditions can alter the expression of biosynthetic gene clusters and therefore the structural diversity and quantity of secondary metabolites produced by microorganisms, including fungi (Bills et al., 2008). For example, growing Fusarium tricinctum on Rice medium supplemented with fruit and vegetable juices led to the discovery of Fusarielin J (Hemphill et al., 2017) while growing Asteromyces cruciatus on Czapek-Dox medium with an altered nitrogen source led to the discovery of Lajollamide A (Gulder et al., 2012). ...
