Search Results (45)

Heavy metal contamination in soil, especially cadmium (Cd) and lead (Pb), poses serious environmental and health risks, particularly in mining regions. While this contamination affects most organisms present in such areas, some filamentous fungi proliferate and immobilize metals in contaminated areas. In this work, six filamentous fungi tolerant to high concentrations of these metals were identified by macroscopic and microscopic morphological characteristics, as well as molecularly, through conserved regions of internal transcribed spacers (ITSs). Tolerance to Cd and Pb was evaluated in solid and liquid culture media, and half the maximum inhibitory concentration (IC₅₀) was assessed. Pb tolerance was observed in Penicillium simplicissimum, Paecilomyces lilacinus, and Rhizopus microsporus (IC₅₀: 3874, 1176, and 211.80 mg/L). Cd tolerance was also noted in Paecilomyces lilacinus, Fusarium oxysporum, Rhizopus microsporus, and Cunninghamella sp. (IC₅₀: 311, 223, 29.25, and 25.18 mg/L). These findings indicate that these fungi have adopted effective strategies for survival in contaminated environments and emphasize their potential for future applications in the bioremediation of multi-metal-contaminated soils. This research lays the groundwork for exploring tolerance mechanisms and evaluating the efficacy of native fungal isolates in mitigating heavy metal contamination. Full article

The fungal genus Cunninghamella, ubiquitously distributed as saprotrophic organisms with occasional endophytic and phytopathogenic manifestations, holds significant biomedical and biochemical importance. During systematic investigations of fungal biodiversity in China, seven novel taxa (C. amphispora sp. nov., C. cinerea sp. nov., C. flava sp. nov., C. hainanensis sp. nov., C. rhizoidea sp. nov., C. simplex sp. nov., and C. yunnanensis sp. nov.) were delineated through integrated taxonomic approaches incorporating morphological characterization, multilocus phylogenetic analyses (ITS-LSU-TEF1α), and physiological assessments. Phylogenetic reconstructions positioned these novel species within a well-supported clade along with C. bainieri and C. verticillata. New species and their diagnostic features are C. amphispora, exhibiting dimorphic sporangiola production; C. cinerea, distinguished by gray pigmentation in the colony; C. flava, characterized by a yellow colony; C. hainanensis and C. yunnanensis, bearing eponymous designations reflecting their geographic origins; and C. simplex, displaying simplified sporangiophore branching. Comprehensive taxonomic descriptions accompanied by photomicrographic illustrations are provided herein. This study constitutes the seventh installment in an ongoing series elucidating early-diverging fungal diversity in China, expanding the global Cunninghamella taxonomic inventory to 63 species and advancing our understanding of mucoralean phylogeny. Full article

Microbial cultures repurposing organic industrial residues for value-added metabolite production is pivotal for sustainable resource use. Highlighting polyunsaturated fatty acids (PUFAs), particularly gamma-linolenic acid (GLA), renowned for their nutritional and therapeutic value. Notably, Zygomycetes’ filamentous fungi harbor abundant GLA-rich lipid content, furthering their relevance in this approach. In this study, the strain C. elegans NRRL Y-1392 was evaluated for its capability to metabolize glycerol and produce lipids rich in GLA under different culture conditions. Various carbon-to-nitrogen ratios (C/N = 11.0, 110.0, and 220.0 mol/mol) were tested in batch-flask cultivations. The highest GLA production of 224.0 mg/L (productivity equal to 2.0 mg/L/h) was observed under nitrogen excess conditions, while low nitrogen content promoted lipid accumulation (0.59 g of lipids per g of dry biomass) without yielding more PUFAs and GLA. After improving the C/N ratio at 18.3 mol/mol, even higher PUFA (600 mg/L) and GLA (243 mg/L) production values were recorded. GLA content increased when the fungus was cultivated at 12 °C (15.5% w/w compared to 12.8% w/w at 28 °C), but productivity values decreased significantly due to prolonged cultivation duration. An attempt to improve productivity by increasing the initial spore population did not yield the expected results. The successful scale-up of fungal cultivations is evidenced by achieving consistent results (compared to flask experiments under corresponding conditions) in both laboratory-scale (Working Volume—Vw = 1.8 L; C/N = 18.3 mol/mol) and semi-pilot-scale (Vw = 15.0 L; C/N = 110.0 mol/mol) bioreactor experiments. To the best of our knowledge, cultivation of the fungus Cunninghamella elegans in glycerol-based substrates, especially in 20 L bioreactor experiments, has never been previously reported in the international literature. The successful scale-up of the process in a semi-pilot-scale bioreactor illustrates the potential for industrializing the bioprocess. Full article

Cheese whey (CW), a byproduct resulting from dairy processing, requires proper treatment and disposal. The use of microalgae during tertiary treatment emerges as a promising option due to its efficiency to remove chemical oxygen demand (COD), total nitrogen (TN), and total phosphorus (TP) from effluents obtained after secondary treatment processes. The present study was focused on treating CW effluent at two organic loads (C₁ and C₂—with different concentrations of COD, TN, and TP) (550–2200 mg·L⁻¹ of COD, 14–56 mg·L⁻¹ of TN, and 4.5–18 mg·L⁻¹ of TP) using the microalga Tetradesmus obliquus alone (C_M1 and C_M2) and in a consortium with the fungus Cuninghamella echinulata (C_MF1 and C_MF2), evaluating the residual values of COD, TN, and TP and removal efficiency. The experiments were carried out in an open system with a volumetric replacement ratio (VRR) of 40 and 60%. The C_M treatment showed residual values of COD in the range of 190–410 mg·L⁻¹ (removal efficiency: 57–68%), TN in the range of 6–24 mg·L⁻¹ (removal efficiency: 29–35%), and TP in the range of 0.90–3.0 mg·L⁻¹ (removal efficiency: 65–68%), after 7 days of volumetric replacement time (VRT) in a semicontinuous mode. In contrast, the consortium (C_MF) showed greater stability and efficiency in contaminant removal compared to the treatment system containing only the microalga, showing residual values of COD in the range of 61–226 mg·L⁻¹ (removal efficiency: 75–77%), TN in the range of 1.8–9.5 mg·L⁻¹ (removal efficiency: 70–74%), and TP in the range of 0.6–3.5 mg·L⁻¹ (removal efficiency: 66–70%), applying a lower VRT of 3 days and reaching the legislation standard for discharge to C_MF1 (VRR: 40 and 60%) and C_MF2 (VRR: 40%). The cell dry weight of 290–850 mg·L⁻¹ was obtained (microalga and microalga–fungus cultivation), which can be a valuable biomass for biotechnological applications. Finally, during microalga–fungus co-cultivation, there was greater system buffering (with less pH variation), ensuring a better system stability. Full article

The Cunninghamella genus has been utilized for the production of PUFA-rich lipids. Therefore, we investigate the impact of plant oil supplementation in the culture medium (soybean oil, rice bran oil, and perilla oil), selected based on their different fatty acid predominant, on lipid production and fatty acid composition in C. elegans (TISTR 3370). All oils significantly boosted fungal growth, each influencing distinct patterns of lipid accumulation within the cells. The cells exhibited distinct patterns of lipid accumulation, forming intracellular lipid bodies, influenced by the different oils. Monounsaturated fatty acids (MUFAs) were found to be the most abundant, followed by polyunsaturated fatty acids (PUFAs) and saturated fatty acids (SFAs) in the fungal lipid cultures. Oleic acid was identified as the primary MUFA, while palmitic acid was the predominant SFA in perilla oil supplements. Remarkably, perilla oil supplement provided the highest total lipid production with arachidonic acid being exclusively detected. The percentage of PUFAs ranged from 12% in the control to 33% in soybean oil, 32% in rice bran oil, and 61% in perilla oil supplements. These findings offer valuable opportunities for advancing biotechnological applications in lipid production and customization, with implications for food and nutrition as well as pharmaceuticals and cosmetics. Full article

Cunninghamella elegans NRRL-1393 is an oleaginous fungus able to synthesize and accumulate unsaturated fatty acids, amongst which the bioactive gamma-linolenic acid (GLA) has potential anti-cancer activities. C. elegans was cultured in shake-flask nitrogen-limited media with either glycerol or glucose (both at ≈60 g/L) employed as the sole substrate. The assimilation rate of both substrates was similar, as the total biomass production reached 13.0–13.5 g/L, c. 350 h after inoculation (for both instances, c. 27–29 g/L of substrate were consumed). Lipid production was slightly higher on glycerol-based media, compared to the growth on glucose (≈8.4 g/L vs. ≈7.0 g/L). Lipids from C. elegans grown on glycerol, containing c. 9.5% w/w of GLA, were transformed into fatty acid lithium salts (FALS), and their effects were assessed on both human normal and cancerous cell lines. The FALS exhibited cytotoxic effects within a 48 h interval with an IC50 of about 60 μg/mL. Additionally, a suppression of migration was shown, as a significant elevation of oxidative stress levels, and the induction of cell death. Elementary differences between normal and cancer cells were not shown, indicating a generic mode of action; however, oxidative stress level augmentation may increase susceptibility to anticancer drugs, improving chemotherapy effectiveness. Full article

The synthetic cytokinin forchlorfenuron (FCF), while seemingly presenting relatively low toxicity for mammalian organisms, has been the subject of renewed scrutiny in the past few years due to its increasing use in fruit crops and potential for bioaccumulation. Despite many toxicological properties of FCF being known, little research has been conducted on the toxicological effects of its secondary metabolites. Given this critical gap in the existing literature, understanding the formation of relevant FCF secondary metabolites and their association with mammalian metabolism is essential. To investigate the formation of FCF metabolites in sufficient quantities for toxicological studies, a panel of four fungi were screened for their ability to catalyze the biotransformation of FCF. Of the organisms screened, Cunninghamella elegans (ATCC 9245), a filamentous fungus, was found to convert FCF to 4-hydroxyphenyl-forchlorfenuron, the major FCF secondary metabolite identified in mammals, after 26 days. Following the optimization of biotransformation conditions using a solid support system, media screening, and inoculation with a solid pre-formed fungal mass of C. elegans, this conversion time was significantly reduced to 7 days—representing a 73% reduction in total reaction time as deduced from the biotransformation products and confirmed by LC-MS, NMR spectroscopic data, as well as a comparison with synthetically prepared metabolites. Our study provides the first report of the metabolism of FCF by C. elegans. These findings suggest that C. elegans can produce FCF secondary metabolites consistent with those produced via mammalian metabolism and could be used as a more efficient, cost-effective, and ethical alternative for producing those metabolites in useful quantities for toxicological studies. Full article

Mucorales are a group of non-septated filamentous fungi widely distributed in nature, frequently associated with human infections, and are intrinsically resistant to many antifungal drugs. For these reasons, there is an urgent need to improve the clinical management of mucormycosis. Miltefosine, which is a phospholipid analogue of alkylphosphocholine, has been considered a promising repurposing drug to be used to treat fungal infections. In the present study, miltefosine displayed antifungal activity against a variety of Mucorales species, and it was also active against biofilms formed by these fungi. Treatment with miltefosine revealed modifications of cell wall components, neutral lipids, mitochondrial membrane potential, cell morphology, and the induction of oxidative stress. Treated Mucorales cells also presented an increased susceptibility to SDS. Purified ergosterol and glucosylceramide added to the culture medium increased miltefosine MIC, suggesting its interaction with fungal lipids. These data contribute to elucidating the effect of a promising drug repurposed to act against some relevant fungal pathogens that significantly impact public health. Full article

Pimavanserin is an approved selective 5-HT_2A receptor inverse agonist for treating Parkinson’s disease psychosis. However, few studies on its metabolism in vitro have been investigated. In this research, eight strains of fungi are used to study the pimavanserin metabolism profiles in vitro and six of them demonstrated positive transformation results. Factors influencing the transformation rate, like substrate concentration, culture time, initial media pH value, culture temperature, and shaking speed, were evaluated and optimized. Cunninghamella blakesleeana AS3.970 provided the best transformation rate of 30.31%, and 10 unreported metabolites were screened by LC-MS/MS. Among these metabolites, M1 is the major one and identified as 1-(4-fluorobenzyl)-3-(4-(2-hydroxy-2-methylpropoxy)benzyl)-1-(1-methylpiperidin-4-yl)urea, which is a hydroxylation product of the pimavanserin. A preliminary molecular docking simulation was performed, which indicated that M1 exhibits similar binding properties with pimavanserin and may become a potential candidate for Parkinson’s disease treatment. Full article

This study aimed to evaluate the microbial oil production by three selected strains, Mortierella isabellina, Cunninghamella echinulata, and Thamnidium elegans, after fermentation of an enzymatic hydrolysate from alkali pre-treated giant reed biomass, in comparison to a semi-synthetic medium, at three increasing nitrogen supplementation levels (0.14, 0.25, and 0.47 g/L). M. isabellina showed the fastest sugar consumption, the highest final cell and oil concentrations (10.9 and 5.6 g/L, respectively), as well as the highest cellular oil content, oil yield/g sugar consumed, and oil productivity (63.4%, 0.19 g/g, and 1 g/L/day, respectively) in the giant reed hydrolysate. The oil yield tended to decrease with an increasing nitrogen content in the cultures. Oleic acid was the most copious fatty acid in the oil for all the fungi. On the contrary, T. elegans exhibited the poorest performances. In particular, M. isabellina showed, respectively, the highest and lowest values of oleic and γ-linolenic acid (52.2 and 3.1%, on average). In comparison, C. echinulata and T. elegans showed much higher γ-linolenic acid content (15.3 and 21.6%, on average). Notably, the C. echinulata cultures showed by far the highest γ-linolenic acid concentration in both substrates (345 and 595 g/L in the giant reed hydrolysate and in the synthetic medium, respectively). Finally, the estimated biodiesel properties of all the oils fell within the limits of the U.S. standards, while the oil of M. isabellina only respected the tighter limits fixed by the E.U. regulations. Full article

Bioconversion of biosynthetic heterocyclic compounds has been utilized to produce new semisynthetic pharmaceuticals and study the metabolites of bioactive drugs used systemically. In this investigation, the biotransformation of natural heterocyclic alkaloid papaverine via filamentous fungi was explored. Molecular docking simulations, using protein tyrosine phosphatase 1B (PTP1B), α-glucosidase and pancreatic lipase (PL) as target enzymes, were performed to investigate the antidiabetic potential of papaverine and its metabolites in silico. The metabolites were isolated from biotransformation of papaverine with Cunninghamella elegans NRRL 2310, Rhodotorula rubra NRRL y1592, Penicillium chrysogeneum ATCC 10002 and Cunninghamella blackesleeana NRRL 1369 via reduction, demethylation, N-oxidation, oxidation and hydroxylation reactions. Seven metabolites were isolated: namely, 3,4-dihydropapaverine (metabolite 1), papaveroline (metabolite 2), 7-demethyl papaverine (metabolite 3), 6,4′-didemethyl papaverine (metabolite 4), papaverine-3-ol (metabolite 5), papaverinol (metabolite 6) and papaverinol N-oxide (metabolite 7). The structural elucidation of the metabolites was investigated with 1D and 2D NMR and mass spectroscopy (EI and ESI). The molecular docking studies showed that metabolite 7 exhibited better binding interactions with the target enzymes PTP1B, α-glucosidase and PL than did papaverine. Furthermore, papaverinol-N-oxide (7) also displayed inhibition of α-glucosidase and lipase enzymes comparable to that of their ligands (acarbose and orlistat, respectively), as unveiled with an in silico ADMET profile, molecular docking and molecular dynamics studies. In conclusion, this study provides evidence for enhanced inhibition of PTP1B, α-glucosidase and PL via some papaverine fungal transformation products and, therefore, potentially better antidiabetic and antiobesity effects than those of papaverine and other known therapeutic agents. Full article

Paeoniflorin is a glycoside compound found in Paeonia lactiflora Pall that is used in traditional herbal medicine and shows various protective effects on the cardio-cerebral vascular system. It has been reported that the pharmacological effects of paeoniflorin might be generated by its metabolites. However, the bioavailability of paeoniflorin by oral administration is low, which greatly limits its clinical application. In this paper, a paeoniflorin-converting enzyme gene (G6046, GenBank accession numbers: OP856858) from Cunninghamella blakesleeana (AS 3.970) was identified by comparative analysis between MS analysis and transcriptomics. The expression, purification, enzyme activity, and structure of the conversion products produced by this paeoniflorin-converting enzyme were studied. The optimal conditions for the enzymatic activity were found to be pH 9, 45 °C, resulting in a specific enzyme activity of 14.56 U/mg. The products were separated and purified by high-performance counter-current chromatography (HPCCC). Two main components were isolated and identified, 2-amino-2-p-hydroxymethyl-methyl alcohol-benzoate (tirs-benzoate) and 1-benzoyloxy-2,3-propanediol (1-benzoyloxypropane-2,3-diol), via UPLC-Q-TOF-MS and NMR. Additionally, paeoniflorin demonstrated the ability to metabolize into benzoic acid via G6046 enzyme, which might exert antidepressant effects through the blood–brain barrier into the brain. Full article

To explore the potential metabolites from rabeprazole sulfide, seven strains of filamentous fungi were screened for their biotransformation abilities. Among these strains, Cunninghamella blakesleeana 3.970 exhibited the best result. Four different culture media were screened in order to identify the most optimal for subsequent research. Single factors such as the initial pH of culture media, culture time, inoculation volume, and media volume were individually investigated to provide the optimum biotransformation conditions. Then, an orthogonal optimization process using a five-factor, four-level L₁₆(4⁵) experiment was designed and performed. Finally, when the substrate concentration is 3 g/L, one major metabolite was detected with a transformation rate of 72.4%. Isolated by semipreparative HPLC, this metabolite was further detected by ESI-MS and NMR. The final data analysis indicated that the metabolite is O-demethylation rabeprazole sulfide. Full article

Renewable and biodegradable materials have attracted broad attention as alternatives to existing conventional plastics, which have caused serious environmental problems. Collagen is a potential material for developing versatile film due to its biosafety, renewability, and biodegradability. However, it is still critical to overcome the low mechanical, antibacterial and antioxidant properties of the collagen film for food packaging applications. To address these limitations, we developed a new technology to prepare composite film by using collagen and fungal-modified APL (alkali pretreatment liquor). In this study, five edible and medical fungi, Cunninghamella echinulata FR3, Pleurotus ostreatus BP3, Ganoderma lucidum EN2, Schizophyllum commune DS1 and Xylariaceae sp. XY were used to modify the APL, and that showed that the modified APL significantly improved the mechanical, antibacterial and antioxidant properties of APL/Collagen composite films. Particularly, the APL modified by BP3, EN2 and XY showed preferable performance in enhancing the properties of the composite films. The tensile strength of the film was increased by 1.5-fold in the presence of the APL modified by EN2. To further understand the effect of fungal-biomodified APL on the properties of the composite films, a correlation analysis between the components of APL and the properties of composite films was conducted and indicated that the content of aromatic functional groups and lignin had a positive correlation with the enhanced mechanical and antioxidant properties of the composite films. In summary, composite films prepared from collagen and fungal biomodified APL showed elevated mechanical, antibacterial and antioxidant properties, and the herein-reported novel technology prospectively possesses great potential application in the food packaging industry. Full article

Natural nitrogen heterocycles biotransformation has been extensively used to prepare synthetic drugs and explore the fate of therapeutic agents inside the body. Herein, the ability of filamentous fungi to biotransform boldine and berberine was investigated. Docking simulation studies of boldine, berberine and their metabolites on the target enzymes: telomerase (TERT) and human protein tyrosine phosphatase 1B (PTP-1B) were also performed to investigate the anticancer and antidiabetic potentials of compounds in silico. The biotransformation of boldine and berberine with Cunninghamella elegans NRRL 2310, Rhodotorula rubra NRRL y1592, Penicillium chrysogeneum ATCC 10002, Cunninghamella blackesleeana MR198 and Cunninghamella blackesleeana NRRL 1369 via demethylation, N- oxidation, glucosidation, oxidation and hydroxylation reactions produced seven metabolites, namely: 1,10-didesmethyl-boldine (1), laurolitsine (2), 1,10-didesmethyl-norboldine (3), boldine-9-O-β-D-glucoside (4), tridesmethyl berberine (5), demethylene berberine (6), and lambertine (7). Primarily, the structures of the metabolites were established by one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) analyses and mass spectrometry. In silico molecular docking simulation of the metabolites of boldine and berberine to the proteins TERT and PTP-1B, respectively, revealed good binding MolDock scores comparable to boldine and berberine and favorable interactions with the catalytic sites of the proteins. In conclusion, this study presented promising biologically prepared nitrogen scaffolds (isoquinolines) of boldine and berberine. Full article