Determination of mycotoxins,alkaloids, phytochemicals,antioxidants and cytotoxicity in Asiatic ginseng (Ashwagandha,Dong quai, <Emphasis Type="Italic">Panax ginseng</Emphasis>)

Mycotoxins and selected hazardous alkaloids in the medicinal plants (Panax ginseng, Angelica sinensis, and Withania somnifera) and dietary supplements were determined. Purine alkaloids were found in majority of samples; however, isoquinoline alkaloids were less abundant than indole. The predominant alkaloids appear to be caffeine (purine group), harman (indole group) and berberine (isoquinoline). Examined medicinal plants and dietary supplements were contaminated by mycotoxins (especially ochratoxin A 1.72–5.83 µg kg^?1), and many species of mold (e.g. Cladosporium, Eurotium, Aspergillus, Rhizopus, Penicillium). MTT cytotoxicity tests revealed that plant and supplements extracts exhibited medium or high cytotoxicity (only Dong quai—low). Moreover, antioxidant activity, total phenolics content and selected phytochemicals were analyzed by spectrophotometric and chromatographic methods. Quercetin and rutin were predominant flavonols (1.94-9.51 and 2.20–7.28 mg 100 g^?1, respectively). Analysis of phenolic acids revealed—gallic acid, as the most abundant, except Panax ginseng, where ferulic acid was prevailing. The results were analyzed by chemometric methods (cluster analysis, ANOVA).     

Use of Mesophilic Fungal Amylases Produced by Solid-state Fermentation in the Cold Hydrolysis of Raw Babassu Cake Starch

Amylases constitute one of the most important groups of industrial enzymes, presenting several applications, such as in the food, textile, and ethanol manufacturing. In this work, a starchy residue from the Brazilian agroindustry, namely babassu cake, was used for the production of amylases by solid-state fermentation and for obtaining sugar hydrolysates, which can be used as building blocks for future bioconversions. Eight filamentous fungi from the genera Aspergillus and Penicillium were screened. Regarding amylase production, A. awamori strains showed well-balanced endoamylase and exoamylase activities, while A. wentii produced an amylolytic complex much richer in the endo-acting enzymes. Simultaneous liquefaction and saccharification using the crude enzyme extracts from the four most promising fungal strains was then investigated applying DOE techniques. The highest total reducing sugar content (24.70 g L^?1) was obtained by the crude extract from A. awamori IOC-3914, corresponding to a hydrolysis yield of 52% of total starch in the cake, while A. awamori IOC-3915 produced the most appropriate extract in terms of glucose release (maximum of 5.52 g L^?1). Multivariate analysis of the DOE studies indicated that these extracts showed their best performance at 50–57 °C under acid conditions (pH 3.6–4.5), but were also able to act satisfactorily under milder conditions (36 °C and pH 5.0), when TRS and glucose released were about 65% of the maximum values obtained. These data confirm the high potential of the enzyme extracts under study for cold hydrolysis of starch.     

Surface enhanced Raman spectroscopy and cultural heritage biodeterioration: Fungi identification in earthen architecture from Paraíba Valley (São Paulo,Brazil)

In this work, Surface Enhanced Raman Spectroscopy (SERS) was employed in the taxonomic identification of fungi found in biofilms formed on earthen architecture walls (adobe, wattle and daub, and rammed earth) of historical buildings in the region known as Paraíba Valley (or São Paulo Historical Valley), which are representative of the first phase of the Brazilian coffee cycle (1820–1880). Very few studies are reported in the literature where SERS-based techniques are used in fungi identification, most of them focused on clinical diagnosis. In the present investigation, pure colonies isolated from biofilms on earthen walls previously identified by classic taxonomy and molecular biology were selected. The genera were Trichoderma, Cladosporium, Aspergillus, Neurospora, Fusarium and Penicillium. The fungi were cultured on solid potato dextrose agar, extracted with ethyl acetate and the extracts were applied on dried Au nanoparticles. The SERS spectra exhibited bands in the 600–1800 cm⁻¹ region which are characteristic of each genus, except Penicillium, as revealed by PCA statistical analysis. This work reports the use of a facile to prepare SERS-active substrate in the identification of microbial communities on earthen architecture walls and is the first step of an investigation aiming at the fast identification of fungi species from biofilms formed on earthen architecture buildings without the need of isolating the pure cultures.     

Inactivation of mildew in rough rice and wheat by gamma irradiation

Rough rice and wheat were irradiated by gamma ray (⁶⁰Co) with different doses and the mildew inactivation efficacy was investigated after 0, 6 and 12 month storage. Five genera of mildew in rough rice and wheat were detected, including Alternaria, Fusarium, Aspergillus, Penicillium and Rhizopus. For Aspergillus, four genera of mold were detected, including Aspergillus Kawachii, Aspergillus glaucus, Aspergillus niger, Aspergillus flavus. Detection rates of the five genera of mildew and four genera of Aspergillus were all reduced with increasing dose after 0, 6 and 12 months storage. The detection rates of the other four genera of mildew had no significant change during storage.     

Overview of fungal lipase: a review

Lipases (triacylglycerolacyl hydrolases, EC3.1.1.3) are class of enzymes which catalyze the hydrolysis of long-chain triglycerides. In this review paper, an overview regarding the fungal lipase production, purification, and application is discussed. The review describes various industrial applications of lipase in pulp and paper, food, detergent, and textile industries. Some important lipase-producing fungal genera include Aspergillus, Penicillium, Rhizopus, Candida, etc. Current fermentation process techniques such as batch, fed-batch, and continuous mode of lipase production in submerged and solid-state fermentations are discussed in details. The purification of lipase by hydrophobic interaction chromatography is also discussed. The development of mathematical models applied to lipase production is discussed with special emphasis on lipase engineering.     

Antimicrobial activity of new bicyclic lactones with three or four methyl groups obtained both synthetically and biosynthetically

Ten new derivatives of isophorone were obtained through a five-step synthesis. Among the products were several unsaturated, bicyclic lactones with three or four methyl groups. These lactones were used as the substrates for biotransformation mediated by selected fungal strains (Fusarium species, Syncephalastrum racemosum, Cunninghamella japonica, Penicillium species, Absidia species, and Pleurotus ostreatus). Four new hydroxylactones were obtained as a result of biotransformation. Because the unsaturated lactone with four methyl groups was a diastereoisomeric mixture, a structural analysis was conducted. The hydroxylactones were also included in this analysis. Both the unsaturated lactones and hydroxylactones were examined for their antimicrobial activity. It was found that some of these compounds exhibited growth inhibition against pathogenic strains of bacteria (Staphylococcus aureus, Pseudomonas fluorescens), yeasts (Candida albicans) and filamentous fungi (Alternaria sp., Penicillium sp.). All obtained compounds were also subjected to scent analysis.     

Rapid and Sensitive Method for Citrinin Determination Using High-Performance Liquid Chromatography with Fluorescence Detection

Citrinin is a toxic product of secondary metabolism of fungi, such as certain Aspergillus, Penicillium, and Monascus species that are usually contaminating cereals. A new sensitive liquid chromatographic method with fluorescence detection was developed, validated, and applied for citrinin determination. The method is based on reversed-phase separation at pH 2.5, where citrinin exhibits the highest fluorescence quantum yield. In this setup, no derivatization step is needed. The method shows linearity in the range between 0.2 μg/mL and 0.1 mg/mL. The detection limit reached is 90 ng/mL (3.6 × 10^?7 M). Validated method was successfully applied on analysis of spiked and real cereal samples.     

Cyclic voltammetry as a screening tool for the fungal degradation of 2,4,6-trinitrotoluene in aqueous media

The use of an electrochemical technique based on cyclic voltammetry (CV) that can be implemented to monitor the degradation of the important nitroaromatic explosive 2,4,6-trinitrotoluene (TNT) in aqueous media when in contact with a microorganism is presented. The microorganism used in this study was an isolated fungus belonging to the Aspergillus genus. In the degradation experiments, during the first 21 days of exposure of the fungal growth to a 68 mg L^?1 TNT aqueous stock solution, the analyte concentration decreased by 44% to 38 mg L^?1 before plateauing through the 58th day of the study. Statistical figures of merit of the technique included linearity in the 1–92 mg L^?1 range and correlation coefficients above 98% for the three reduction peaks of TNT, a limit of detection (LOD) of 3 mg L^?1, a limit of quantification of 10 mg L^?1 and a method precision of 3.8% relative standard deviation (%RSD). Day-to-day and week-to-week repeatability were low at 5.1%RSD and 5.8%RSD, respectively. The results herein exhibit first-order kinetics for the ‘ortho’ nitro group. A clear to yellow colour transition in the control solution and fungi samples suggests the appearance of a TNT metabolite. UV-Vis spectrophotometry supports the presence of a possible derivative of TNT in the fungi samples.     

Solid-State Fermentation of Rapeseed Meal with the White-Rot Fungi Trametes versicolor and Pleurotus ostreatus

Rapeseed meal is valuable high-protein forage, but its nutritional value is significantly reduced by the presence of a number of antinutrients, including phenolic compounds. Solid-state fermentation with white-rot fungi was used to decrease the sinapic acid concentration of rapeseed meal. After 7 days of growth of Trametes versicolor and Pleurotus ostreatus, the sinapic acid content of rapeseed meal was reduced by 59.9 and 74.5 %, respectively. At the end of the experiment, sinapic acid concentration of T. versicolor cultures decreased by 93 % of the initial value; in the case of cultures of P. ostreatus, 93.2 % reduction was observed. Moreover, cultivation of white-rot fungi on rapeseed meal resulted in the intensive production of extracellular laccase, particularly strong during the late phases of growth of T. versicolor. The obtained results confirm that both fungal species may effectively be used to decompose antinutritional phenolics of rapeseed meal. Rapeseed meal may also find use as an inexpensive and efficient substrate for a biotechnological production of laccase by white-rot fungi.     

Data analysis of pyrolysis—chromatograms by means of simca pattern recognition

The variability between repetitive pyrolysis—chromatograms of the same type of samples is not totally random, but can in part be modelled by a principal components (PC) model. This makes it possible to use efficiently repetitive pyrolysis-chromatograms of samples of known types to obtain separate PC models for each type. Samples of unknown origin can then be classified according to which of the PC models their pyrolysis-chromatograms are most similar.The methodology is illustrated using pyrolysis—gas chromatograms of two species of the fungal genus Penicillium.     

Isolation,Purification, and Characterization of Antimicrobial Compound 6-[1,2-dimethyl-6-(2-methyl-allyloxy)-hexyl]-3-(2-methoxy-phenyl)-chromen-4-one from Penicillium sp. HT-28

A fungal culture (Penicillium sp., HT-28), isolated from soil has been evaluated for its bioactivity, which showed broad spectrum antimicrobial activity and was effective against methicillin-resistant Staphylococcus aureus (MRSA) also. Statistical optimization of the medium by response surface methodology (RSM) enhanced the antimicrobial activity up to 1.8-fold. Column chromatography was used to isolate the active compound (A), which was characterized to be 6-[1,2-dimethyl-6-(2-methyl-allyloxy)-hexyl]-3-(2-methoxy-phenyl)-chromen-4-one by various spectroscopic techniques such as infrared (IR), ¹H and ¹³C nuclear magnetic resonance (NMR) spectra, and mass spectroscopy. Minimum inhibitory concentration (MIC) of the active compound (A) ranged from 0.5 to 15 μg/mL. Viable cell count studies of the active compound (A) showed S. aureus, Escherichia coli, Staphylococcus epidermidis, and Salmonella typhimurium 1 to be the most sensitive. The compound retained its bioactivity after treating it at 100 °C for 1 h. Furthermore, the compound (A) when tested for its biosafety was found neither to be cytotoxic nor mutagenic. The study demonstrated that an apparently novel compound isolated from Penicillium sp. (HT-28) seems to be a stable and potent antimicrobial.     

Abiotic and biotic degradation of oxo-biodegradable polyethylenes

Conventional polymeric materials accumulate in the environment due to their low biodegradability. However, an increase in the biodegradation rate of these polymers may be obtained with the addition of pro-degrading substances. This study aimed to evaluate abiotic and biotic degradation of polyethylenes (PEs) using plastic bags of high-density polyethylene (HDPE) and linear low-density polyethylene (LLDPE) formulated with pro-oxidant additives as test materials. These packaging materials were exposed to natural weathering and periodically analyzed with respect to changes in mechanical and structural properties. After a year of exposure, residue samples of the bags were incubated in substrates (compost of urban solid waste, perlite and soil) at 58 °C and at 50% humidity. The biodegradation of the materials was estimated by their mineralization to CO₂. The molar mass of the pro-oxidant-activated PE decreased and oxygen incorporation into the chains increased significantly during natural weathering. These samples showed a mineralization level of 12.4% after three months of incubation with compost. Higher extents of mineralization were obtained for saturated humidity than for natural humidity. The growth of fungi of the genera Aspergillus and Penicillium was observed on PE films containing pro-oxidant additives exposed to natural weathering for one year or longer. Conventional PE films exposed to natural weathering showed small biodegradation.     

Arsenic species in terrestrial fungi and lichens from Yellowknife,NWT, Canada

Levels of total arsenic and arsenic species were determined in fungi collected from Yellowknife, NWT, Canada, an area that has been affected by past mining activities and elevated arsenic levels. Lichens (belonging to Cladonia and Cladina genera), as well as the mushrooms Coprinus comatus, Paxillus involutus, Psathyrella candolleana and Leccinum scabrum, were studied for the first time. Most of the fungi contained elevated arsenic levels with respect to data found in the literature for background levels. Minor amounts of arsenobetaine were found in all lichen samples. The major water‐soluble arsenic species in the fungi were inorganic arsenic for lichens and Psathyrella candolleana, arsenobetaine for Lycoperdon pyriforme and Coprinus comatus, and dimethylarsenate for Paxillus involutus and Leccinum scabrum. A large proportion of water‐soluble arsenic in Paxillus involutus occurred as an unknown compound, which did not co‐chromatograph with any of the available standard arsenic compounds. Low proportions of water‐soluble arsenic species (made evident by low extraction efficiencies) were observed in the majority of fungi studied. Arsenic that is not extracted may be bound to lipids, cell components or proteins, or might exist on the surface of the fungus as minerals. Copyright © 2000 John Wiley & Sons, Ltd.     

Degradation study of carbamide cellulose-based macroporous resin

Degradation of macroporous resin of TDI cross-linked cellulose named TCC prepared by crosslinking cellulose with TDI was investigated. The life span for 70 % weight loss, which was estimated based on the changes in dynamic parameters obtained from thermogravimetric analysis, was 74,507 h. Three bacteria including Trametes gallic, Gloeophyllum trabeum, Aspergillus, which were separated and purified from nature, were used to decompose macroporous resin solely or in combination. The life span was reduced to 720 h when combination of Trametes gallic, Gloeophyllum trabeum, Aspergillus was used to decompose TCC at 25 °C, relative humidity of 70 % and pH = 5. Also, the influence of carbon content, nitrogen content and pH value on the biodegradation rate of TCC was discussed. Degradation rate of TCC was highest when mass ratio of glucose to TCC was 0.04, mass ratio of nitrogen to TCC was 0.04 and pH value was 5.     

Chemical constituents of the fermentative extracts of marine fungi Phoma sp. CZD-F11 and Aspergillus sp. CZD-F18 from Zhoushan Archipelago,China

A new diphenyl ether (1) as well as 20 other compounds were identified from the fermentative extracts of marine-derived fungi Phoma sp. CZD-F11 (Compounds 1–8) and Aspergillus sp. CZD-F18(Compounds 9–21). Their structures were elucidated on the basis of extensive spectroscopic analysis. The broth extracts of the fungi exhibited very good anticancer activity against H1975 cells with 5.62 and 25.8% viability at concentration of 10 μg/mL for Phoma sp. CZD-F11 and Aspergillus sp. CZD-F18, respectively. The inhibitory activity of all compounds against PC-3 cell lines, BRD4 and aromatase were evaluated. The results showed compound 7 exhibited moderate anticancer activity with 66.1% inhibition against PC-3 cell lines at the concentration of 10 μg/mL. Compound 7 and 8 exhibited favourable BRD4 inhibitory activity with 78.5 and 76.4% inhibition at the concentration of 10 μg/mL.     

Species-specific optical genosensors for the detection of mycotoxigenic Fusarium fungi in food samples

Plant-pathogenic Fusarium species, Fusarium verticillioides and Fusarium proliferatum, are the major producers of fumonisins which are one of the most common mycotoxins found in maize. Herein, we report the development of specific and sensitive genosensors for detecting these two closely related Fusarium species in food samples. The sensors are based on species-specific capture and detection probes, which bind to the intergenic spacer region of rDNA (IGS). Oligonucleotide functionalized magnetic microbeads are used to capture the target DNA which is then detected using biotinylated detection probes and a streptavidin-coupled label. The developed genosensors had detection limits of 1.8 pM and 3.0 pM for F. proliferatum and F. verticillioides, respectively, using synthetic DNA targets. Furthermore, the biosensors were used to analyze natural fungal contamination of commercial maize samples. After amplification of the genomic DNA the sensors detected the presence of the fungi, in accordance with previous results obtained with PCR. No cross-reactivity between F. verticillioides and F. proliferatum, or other fungi species tested, was observed. The developed biosensors can provide a valuable tool to evaluate the potential for mycotoxin contamination in conditions where detection of mycotoxins directly is challenging.     

A straight forward and first total synthesis of Penilumamides B–D

A simple and straight forward first total synthesis of rare lumazine peptides, Penilumamide B, C and D isolated from the marine-derived fungi Aspergillus and Penicillium sp. is described from a common starting material, 1,3-dimethyllumazine-6-carboxylic acid. Penilumamide C was prepared from Penilumamide B by oxidation of the methionine residue to the corresponding sulphone.     

Synergized Antimicrobial Activity of Eugenol Incorporated Polyhydroxybutyrate Films Against Food Spoilage Microorganisms in Conjunction with Pediocin

Biopolymers and biopreservatives produced by microorganisms play an essential role in food technology. Polyhydroxyalkanoates and bacteriocins produced by bacteria are promising components to safeguard the environment and for food preservation applications. Polyhydroxybutyrate (PHB)-based antimicrobial films were prepared incorporating eugenol, from 10 to 200 μg/g of PHB. The films were evaluated for antimicrobial activity against foodborne pathogens, spoilage bacteria, and fungi such as Staphylococcus aureus, Escherichia coli, Salmonella typhimurium, Bacillus cereus, Aspergillus flavus, Aspergillus niger, Penicillium sp., and Rhizopus sp. The synergistic antimicrobial activity of the films in the presence of crude pediocin was also investigated. The broth system containing pediocin (soluble form) as well as antimicrobial PHB film demonstrated an extended lag phase and a significant growth reduction at the end of 24 h against the bacteria. Crude pediocin alone could not elicit antifungal activity, while inhibition of growth and sporulation were observed in the presence of antimicrobial PHB film containing eugenol (80 μg/g) until 7 days in the case of molds, i.e., A. niger, A. flavus, Penicillium sp., and Rhizopus sp. in potato dextrose broth. In the present study, we identified that use of pediocin containing broth in conjunction with eugenol incorporated PHB film could function in synergized form, providing effective hurdle toward food contaminating microorganisms. Furthermore, tensile strength, percent crystallinity, melting point, percent elongation to break, glass transition temperature, and seal strength of the PHB film with and without eugenol incorporation were investigated. The migration of eugenol on exposure to different liquid food simulants was also analyzed using Fourier transform infrared spectroscopy. The study is expected to provide applications for pediocin in conjunction with eugenol containing PHB film to enhance the shelf life of foods in the food industry.     

Protease Produced by Endophytic Fungi: A Systematic Review

The purpose of this systematic review was to identify the available literature of production, purification, and characterization of proteases by endophytic fungi. There are few complete studies that entirely exhibit the production, characterization, and purification of proteases from endophytic fungi. This study followed the PRISMA, and the search was conducted on five databases: PubMed, PMC, Science Direct, Scopus Articles, and Web of Science up until 18 May 2021, with no time or language restrictions. The methodology of the selected studies was evaluated using GRADE. Protease production, optimization, purification, and characterization were the main evaluated outcomes. Of the 5540 initially gathered studies, 15 met the inclusion criteria after a two-step selection process. Only two studies optimized the protease production using statistical design and two reported enzyme purification and characterization. The genus Penicillium and Aspergillus were the most cited among the eleven different genera of endophytic fungi evaluated in the selected articles. Six studies proved the ability of some endophytic fungi to produce fibrinolytic proteases, demonstrating that endophytic fungi can be exploited for the further production of agents used in thrombolytic therapy. However, further characterization and physicochemical studies are required to evaluate the real potential of endophytic fungi as sources of industrial enzymes.     

Isolation and Screening of Microorganisms for R-(+)-Limonene and (−)-β-Pinene Biotransformation

This work is focused on the biotransformation of R-(+)-limonene and (?)-β-pinene to bioflavor production. To carry out the present study, 405 microorganisms were tested for their ability to bioconvert the substrates. From the isolated microorganisms, 193 were selected in the prescreening using mineral medium for limonene degradation. At the screening step, eight strains were able to convert R-(+)-limonene and 15 to transform (?)-β-pinene, both in α-terpineol. The highest concentration in α-terpineol from R-(+)-limonene was about 3,450 mg/L for Penicillium sp. isolated from eucalyptus steam. From (?)-β-pinene, the highest product concentration of 675.5 mg/L was achieved using an Aspergillus sp. strain isolated from orange tree stem.