Penicillium Crustosum Research Articles

Biosynthesis of Xylariolide D in Penicillium crustosum Implies a Chain Branching Reaction Catalyzed by a Highly Reducing Polyketide Synthase.

Fungi are important sources for the discovery of natural products. During the last decades, technological progress and the increasing number of sequenced genomes facilitated the exploration of new secondary metabolites. Among those, polyketides represent a structurally diverse group with manifold biological activities. In this study, we successfully used genome mining and genetic manipulation for functional proof of a polyketide biosynthetic gene cluster from the filamentous fungus Penicillium crustosum. Gene activation in the native host and heterologous expression in Aspergillus nidulans led to the identification of the xil cluster, being responsible for the formation of the 6-methyl-2-pyrone derivative xylariolide D. Feeding with 13C-labeled precursors supported the hypothesis of chain branching during the backbone formation catalyzed by a highly reducing fungal polyketide synthase. A cytochrome P450-catalyzed hydroxylation converts the PKS product to the final metabolite. This proved that just two enzymes are required for the biosynthesis of xylariolide D.

Preparation of Chitosan with High Antibacterial Efficiency from Penicillium crustosum TZ18

Preparation of Chitosan with High Antibacterial Efficiency from Penicillium crustosum TZ18

POTENTIAL OF Penicillium crustosum METABOLITES IN CONTROLLING PARTHENIUM WEED

Parthenium (Parthenium hysterophorus L.) ranks amongst the top ten most devastating weeds. It has spread on a large scale in various regions of Punjab and Khyber Pakhtunkhwa, Pakistan. In an attempt to search alternatives to chemical herbicides for control of this weed, metabolites of a fungal species Penicillium crustosum were evaluated for their herbicidal potential. The fungus was grown in two growth media viz. potato dextrose broth (PDB) and malt extract broth (MEB) for two weeks. After filtration, the culture filtrates from both the media were used in original (100%) as well as diluted (50%) forms. The experiment was carried out in 9-cm Petri plates, having 25 parthenium seeds in each plate. The effect of metabolites was assessed on germination and seedling growth. PDB metabolites proved more inhibitory against various studied parameters than the metabolites prepared in MEB. There was 62%, 88%, 91% and 93% suppression in germination, shoot length, root length and seedling dry biomass over control due to original metabolites prepared in PDB as compared to 33%, 74%, 94% and 81% reduction in these parameters, respectively, due to MEB metabolites.

Fungal Endophytes of Vitis vinifera-Plant Growth Promoters or Potentially Toxinogenic Agents?

Fungal endophytes occurring in grapevine (Vitis vinifera L.) are usually important sources of various compounds with biological activities with great potential for use in agriculture. Nevertheless, many species isolated from this plant belong to the genera Fusarium, Alternaria, or Aspergillus, all of which are well-known to produce mycotoxins. Our study is focused on the assessment of the toxinogenic potential of fungal endophytes isolated from vineyards in the Czech Republic. In total, 20 endophytic fungal species were cultivated in wine must, and 57 mycotoxins of different classes were analysed by liquid chromatography coupled with mass spectrometry. As a result, alternariol, tentoxin, meleagrin, roquefortine C, gliotoxin, and verruculogen were detected in the culture medium, of which verruculogen followed by gliotoxin were the most frequent (present in 90 and 40% of samples, respectively) and most concentrated (up to thousands ng/mL). The alternaria mycotoxins alternariol and tentoxin were detected not only in Alternaria sp. cultures, but traces of these mycotoxins were also quantified in the Diatripe and Epicoccum cultures. Meleagrin and roquefortine C were detected in Didymella sancta and Penicillium crustosum, gliotoxin was detected in Alternaria sp., Didymella sp., Aureobasidium pullulans, Cladosporium herbarum, Penicillium crustosum and Pleurophoma ossicola, and verruculogen was quantified in 99% of endophytic isolates investigated. The potential of endophytes to produce mycotoxins should be carefully checked, specifically in cases where they are intended for the purpose of V. vinifera growing.

Facile Synthesis and Size Distribution Analysis of Silver Nanoparticles of Fungus Penicillium crustosum, Isolated from Taxus baccata Linn.

The present study was aimed at the synthesis and size distribution analysis of silver nanoparticles (AgNPs), from a fungal endophyte Penicillium crustosum. The fungus was isolated from the needles of Taxus baccata Linn. and identified as Penicillium crustosum by molecular characterization. The fungal extract of P. crustosum was subjected to the synthesis of AgNPs. UV-vis-spectroscopy confirmed the formation of AgNPs by forming the specific peaks at 390-450 nm. The lyophilized AgNPs were subjected to size distribution analysis using tools like X-ray diffraction, energy dispersive spectroscopy (EDS), field emission scanning electron microscopy (FEG-SEM), and high-resolution transmission electron microscopy (HRTEM). The average crystalline size of AgNPs was calculated by Debye-Scherrer’s formula which showed AgNPs of 36.93 nm. These findings substantiated the results of FEG-SEM and HRTEM, where the average size was below 50 nm. Dynamic light scattering results showed Z-average of AgNPs as 352.0 rnm with 0.982 PdI. Zeta potential was -19.1 mV which showed the capacity of formed AgNPs to decline agglomeration. EDS analysis proved that the requisite phase of silver is present in the sample. The present study displayed the potential of P. crustosum as reducing agent which converted silver ions into AgNPs in eco-friendly and cost-effective manner.

Formation of 3-Orsellinoxypropanoic Acid in Penicillum crustosum is Catalyzed by a Bifunctional Nonreducing Polyketide Synthase.

The heterologous expression of a nonreducing polyketide synthase gene oesA from Penicillium crustosum led to the identification of orsellinoylpropanoic acid (1). Domain deletion and recombination proved that OesA catalyzes not only the formation of orsellinic acid but also its transfer to 3-hydroxypropanoic acid. Both ACP domains contribute independently and complementarily to the product formation. Feeding experiments provided evidence that only the orsellinyl residue is derived from acetate.

Linear polyketides produced by co-culture of Penicillium crustosum and Penicillium fellutanum.

The online version contains supplementary material available at 10.1007/s42995-021-00125-8.

Bioactivities of six plant essential oils against some isolated microbes from an archaeological limestone statue at the Saqqara excavation

The effect of microbial activity on the deterioration of archaeological stone items is a worldwide issue, and conserving them with low-toxicity, ecologically benign and naturally biocides is a difficult undertaking. Molecular identification of the microbial communities from a deteriorated archaeological object (Ptahshepses stone statue) located from the Saqqara excavation, Giza, Egypt was recorded. Six essential oils (EOs) (black cumin, clove, geranium, lavender, lemongrass, and thyme) were tested for antimicrobial activity against six degrading fungal and bacterial species. Alternaria alternate, Aspergillus flavus, A. niger, Cladosporium halotolerans, Penicillium crustosum, and Trichoderma viride and three bacterial species, Pseudomonas protegens, P. putida, and Serratia odorifera, were isolated. Of the most effective EOs, thyme showed the highest inhibition percentage (143.4%) against Serratia odorifera, followed by P. putida (135%), and Pseudomonas protegens (131.5%). Lemongrass and clove EOs had minimum inhibitory concentrations (MICs) ranging from 0.5 µL/mL to 2 µL/mL for all isolated deteriorated fungal and bacterial species, while the lowest efficiency EOs were lavender, geranium and black cumin. It can be concluded that thyme and lemongrass EOs have a potential use for protecting the Ptahshepses stone statue from microbial deterioration.

A mixed culture of Propionibacterium freudenreichii and Lactiplantibacillus plantarum as antifungal biopreservatives in bakery product

In this study, 15 strains of Lactic acid bacteria (Allata et al.) and 6 strains of propionic acid bacteria (PAB) were firstly tested for their antifungal activity against three spoilage fungi, Aspergillus niger, Penicillium crustosum and Aspergillus flavus. Two strains of LAB and PAB were selected and assessed, alone or paired, for their abilities to inhibit the most resistant mold, Aspergillus niger. A mixed culture of P. freudenreichii D6 and L. plantarum L9 was found to be the most active and their optimal inoculum was 1 × 108 and 1 × 106 CFU/mL, respectively. Furthermore, the in situ antifungal effect of the mixed culture was evaluated against bakery product spoilage fungi. It was found that the growth of airborne fungi was delayed for up to 7 days. The cell number of L. plantarum L9 was slightly increased by the presence of P. freudenreichii D6. Quantification of three main acids (propionic, lactic and acetic acid) produced by the protective cultures suggested that acetic acid was mainly responsible for the antifungal activity. In conclusion, the mixed culture of P. freudenreichii and L. plantarum may be exploited as biopreservatives in bakery products.

Biological potential and microencapsulation of Cinnamomum cassia essential oil as an alternative for pest control in stored maize

This study proposes a review of biological potential of cinnamon (Cinnamomum cassia) essential oil with a focus on microencapsulation as an alternative to control the occurrence of pests in stored maize grains. Due to the demands on corn productivity, there is the need to improve grain storage processes and conditions, since that in this stage there are quantitative and/or qualitative losses, mainly due to the maize weevils (Sitophilus zeamais) and the incidence of mycotoxin-producing fungi (Penicillium crustosum, Alternaria alternata and Aspergillus flavus). The control of these pests is usually carried out with chemical insecticides, which can leave toxic residues in the grain. Therefore, the microencapsulation of essential oils appears as a promising alternative, considering the volatility of aromatic compounds, which are largely responsible for the activity against pests.

Conversion of viridicatic acid to crustosic acid by cytochrome P450 enzyme-catalysed hydroxylation and spontaneous cyclisation

Cytochrome P450 monooxygenases (P450s) are considered nature’s most versatile catalysts and play a crucial role in regio- and stereoselective oxidation reactions on a broad range of organic molecules. The oxyfunctionalisation of unactivated carbon-hydrogen (C-H) bonds, in particular, represents a key step in the biosynthesis of many natural products as it provides substrates with increased reactivity for tailoring reactions. In this study, we investigated the function of the P450 enzyme TraB in the terrestric acid biosynthetic pathway. We firstly deleted the gene coding for the DNA repair subunit protein Ku70 by using split marker-based deletion plasmids for convenient recycling of the selection marker to improve gene targeting in Penicillium crustosum. Hereby, we reduced ectopic DNA integration and facilitated genetic manipulation in P. crustosum. Afterward, gene deletion in the Δku70 mutant of the native producer P. crustosum and heterologous expression in Aspergillus nidulans with precursor feeding proved the involvement of TraB in the formation of crustosic acid by catalysing the essential hydroxylation reaction of viridicatic acid.Key points•Deletion of Ku70 by using split marker approach for selection marker recycling.•Functional identification of the cytochrome P450 enzyme TraB.•Fulfilling the reaction steps in the terrestric acid biosynthesis.

Penicillium and Talaromyces Species as Postharvest Pathogens of Pear Fruit (Pyrus communis) in Serbia.

Pears are one of the oldest and the third most important fruit species grown in temperate regions. They are consumed because of their nutritional and health benefits, in fresh form or as various processed products. This article resolves the etiology of the Penicillium-like mold symptoms on pear fruits in Serbia. Samples of pear fruits with blue mold and other Penicillium-like mold symptoms were collected in Serbia from 2016 to 2019, from four storages. The recovered isolates were identified and characterized according to a polyphasic approach. Morphological and physiological analyses were performed on three media and five temperatures, respectively. Four loci (internal transcribed spacer, beta-tubulin, calmodulin, and DNA-dependent RNA polymerase II second largest subunit) were used for sequencing, genetic identification, and phylogenetic analyses. The results of the identification by conventional and molecular methods were in agreement, and they revealed that the obtained isolates belong to five species: Penicillium crustosum, P. expansum, P. italicum, Talaromyces minioluteus, and T. rugulosus. In a pathogenicity test, P. crustosum, P. expansum, T. minioluteus, and T. rugulosus produced decay on artificially inoculated pear fruits, and P. italicum induced tissue response lesions. The results of this study are the first reports of T. minioluteus and T. rugulosus as postharvest pear pathogens. Also, these are the first world records of T. minioluteus, T. rugulosus, and P. italicum on fruits of European pear. Furthermore, this is the first finding of P. crustosum, P. expansum, P. italicum, T. minioluteus, and T. rugulosus on pear fruit in Serbia.

New derivatives of the iridoid specioside from fungal biotransformation.

Iridoids are widely found from species of Bignoniaceae family and exhibit several biological activities, such as anti-inflammatory, antimicrobial, antioxidant, and antitumor. Specioside is an iridoid found from Tabebuia species, mainly in Tabebuia aurea. Thus, here fungus-mediated biotransformation of the iridoid specioside was investigated by seven fungi. The fungus-mediated biotransformation reactions resulted in a total of nineteen different analogs by fungus Aspergillus niger, Aspergillus flavus, Aspergillus japonicus, Aspergillus terreus, Aspergillus niveus, Penicillium crustosum, and Thermoascus aurantiacus. Non-glycosylated specioside was the main metabolite observed. The other analogs were yielded from ester hydrolysis, hydroxylation, methylation, and hydrogenation reactions. The non-glycosylated specioside and coumaric acid were yielded by all fungi-mediated biotransformation. Thus, fungus applied in this study showed the ability to perform hydroxylation and glycosidic, as well as ester hydrolysis reactions from glycosylated iridoid. KEY POINTS: • The biotransformation of specioside by seven fungi yielded nineteen analogs. • The non-glycosylated specioside was the main analog obtained. • Ester hydrolysis, hydroxylation, methylation, and hydrogenation reactions were observe.

Bioremediation of an agricultural saline soil contaminated with endosulfan and Escherichia coli by an active surface agent induced in a Penicillium crustosum culture

This study evaluates the production of a biological active surface agent (BASA) through its surface tension (ST) and emulsifying activity (E24) for endosulfan degradation (ED) and Escherichia coli growth inhibition (EcGI) in an agricultural saline soil. The fungus, identified as Penicillium crustosum was isolated from the Citrus sinensis peel (CsP), then the surface properties were evaluated in 9 culture media through a Taguchi L9 experimental design. The culture conditions included: stirring speed, pH, carbon (C) and nitrogen (N) sources; being glucose, NH4N03, 120 rpm and pH of 5, the most significant parameters in the BASA production. The BASA identified as a lipopeptide type, showed a ST = 38 mN m−1 and E24=71%. Both properties were stable at 80 °C, while ST presented stability in the pH range of 2 – 12, and a saline concentration of 200 g L−1; E24 was also stable at a pH between 8–12. Further application of BASA and fungal inoculum to a contaminated agricultural saline soil presented an EcGI of 99.8% on the 8th day, and ED of 92.9 ± 4.7% in 30 days, respectively; being the first report that uses this fungus for pesticide and bacteria elimination from an agricultural saline soil.

Mycoremediation of an agricultural salty soil contaminated with endosulfan by Penicillium crustosum: and agronomic bioassays with Phaseolus leptostachyus

The fungus Penicillium crustosum was employed for endosulfan biodegradation, finding that sulphate endosulfan and mono alcohol endosulfan were the main compounds produced; therefore, an oxidative degradation pathway was suggested. A 93 ± 4.7% of Endosulfan degradation after one month of treatment of a highly salty agricultural soil was obtained, where ΔST was up to 17 ± 0.58 mN m−1, (related to the water value of 72 mNm−1), that was induced by the fungus during soil mycoremediation Additionally, an improvement in soil quality (reduction of clay proportion and salinity, as well as an increase of soluble phosphorus, carbon content and organic matter) was observed during the mycoremediation treatment. The phytotoxicity of the pesticide on Phaseolus leptostachyus was evaluated in the soil without the fungus addition (control), where the pesticide was translocated in the crop, presenting a negative effect in germination index, root length and weight, aerial weight, humidity, and proline content. This contrasted with the effect on the crop grown in the soil treated with P. crustosum, which had better agronomic characteristics. This is first report in which the effect of this property allows the pesticide biodegradation, due to a combined Endosulfan bioavailability and fungal biodegradation.

Precursor Supply Increases the Accumulation of 4-Hydroxy-6-(4-hydroxyphenyl)-α-pyrone after NRPS-PKS Gene Expression.

Expression of a nonribosomal peptide synthetase-nonreducing polyketide synthase hybrid gene pcr10109 from Penicillium crustosum PRB-2 in Aspergillus nidulans led to the accumulation of 4-hydroxy-6-(4-hydroxyphenyl)-α-pyrone (1). Adding para-hydroxybenzoic acid into the medium in which the overexpressing mutant is growing increased the product yield up to 5-fold. This strategy could be helpful for heterologous gene expression experiments requiring special substrates for product formation.

Thymus serpyllum Essential Oil and Its Biological Activity as a Modern Food Preserver.

The aim of this study was to analyze the chemical composition and biological and antibiofilm activity of the essential oil (EO) of Thymus serpyllum with the use of a MALDI-TOF MS Biotyper. The main compounds of the EO were thymol, 18.8%; carvacrol, 17.4%; o-cymene, 15.4%; and geraniol, 10.7%. It was found that free-radical scavenging activity was high. The highest antimicrobial activity was observed against Pseudomonas aeruginosa, Salmonella enteritidis, and biofilm-forming bacteria. The changes in the biofilm structure after T. serpyllum EO application confirmed the inhibitory action and the most pronounced effect was observed on Bacillus subtilis biofilm. The antifungal activity of the vapor phase was the most effective against Penicillium crustosum. T. serpyllum should be a suitable alternative to synthetic antioxidants as well as antimicrobials. The EO of T. serpyllum can be used in the vapor phase in the storage of root vegetables as well as a growth inhibitor of Penicillium on bread.

Eco-friendly bioprocess using agar plug immobilized Penicillium crustosum PWWS-6 biomass for treatment of wastewater contaminated with toxic Congo red dye for use in agriculture

Recycling and valorization of noxious dye contaminated industrial wastewater minimize prevailing agricultural water dearth. In this context, the current manuscript investigates and fabricates a rapid, ecofriendly, and cost effective treatment system using a potent dye degrading microbial agent. An agar plug immobilized Penicillium crustosum PWWS-6, a filamentous fungus isolated from veterinary clinical waste dump, for the first time reported to decolorize (81.86 ± 0.03 %) hazardous diazo dye, (Congo red, 100 ppm) in 24 h. Lignin peroxidase, azo reductase, veratryl alcohol oxidase, and NADH-DCIP reductase enzymes were induced significantly during mycoremediation bioprocess which is affected by incubation time, temperature, pH, inoculum volume, and dye concentration of aqueous medium. The one-factor-at-a-time (OFAT) and response surface methodology (RSM) were adopted to optimize these cultural parameters and generated optimized conditions (incubation time, 16 h; temperature, 27 °C; and inoculum volume, 6 fungal discs), for maximum degradation (99.85 %) of 100 ppm CR at pH 7.0. Further, using OFAT approach, P. crustosum also showed 72.14 % degradation of CR at much higher concentration of 1000 ppm in 72 h. UV–vis spectra verified complete disappearance of dye from colored broth. Fourier Transform Infrared (FTIR) spectroscopy along with Field Emission Scanning Electron Microscopy (FESEM) subsequently confirmed (–N=N–) bond destabilization following degradation. Furthermore, the non-toxic degraded metabolites showed improved percent germination, radical and plumule length of Cicer arietinum and Hordeum vulgare seeds compared with untreated CR contaminated wastewater.

Effect of the Pesticide Endosulfan and Two Different Biostimulants on the Stress Responses of Phaseolus leptostachyus Plants Grown in a Saline Soil

Soil salinity and the indiscriminate use of agrochemicals has significantly reduced the productivity of the ‘Chinampas’ agroecosystem in Mexico City. Crop improvement under these stressful conditions may be achieved by soil bioremediation. In this study, we checked the effects of the organochlorine pesticide endosulfan and bioremediation with Penicillium crustosum or a citric waste on the growth of Phaseolus leptostachyus plants in saline soil from the Chinampas area. Biochemical markers associated with specific stress responses were also determined after one month of growth in the different substrates. Plant growth was stimulated by bioremediation of the soil. Both biostimulants reduced the degree of stress affecting the plants, as shown by the increase in photosynthetic pigments and the reduction of proline, malondialdehyde (MDA), and H2O2 contents, and the activation of antioxidant systems. However, the biostimulants appeared to mitigate oxidative stress through different mechanisms. Endosulfan contamination inhibited seed germination—which was reverted to control values in the presence of the biostimulants—and further decreased plant growth. No clear patterns of variation of biochemical stress markers were observed combining endosulfan and the biostimulants. In any case, bioremediation with P. crustosum and/or citric waste is recommended to improve the germination and growth of P. leptostachyus plants.

Mycobiota dynamics and mycotoxin detection in PGI Salame Piemonte

The complex mycobiota that colonizes traditional fermented sausages plays an important role in the organoleptic properties of such products. The aim of the present study was to investigate fungal diversity and mycotoxin production during maturation of PGI Salame Piemonte. Casing and meat samples were collected at five sampling times from three different batches produced in the same factory and analysed using culture-dependent and independent approaches. Penicillium nalgiovense, which was deliberately inoculated, and Debaryomyces hansenii were the most dominant taxa in casings. Several other fungi mainly belonging to Penicillium crustosum, Penicillium glabrum, Penicillium nordicum, Cladosporium spp., Candida sake, Candida zeylanoides and Yarrowia divulgata were also identified. The casing mycobiota was compared to that of the meat using a metataxonomic approach and a higher fungal diversity was observed in meat as compared to casings. Mycotoxins and penicillin G were monitored using QTOF LC-MS and only trace amounts of roquefortine C were detected in two batches. The present study highlighted the diversity of Salame Piemonte mycobiota and the important contribution of autochthonous fungi to its diversity. The absence of mycotoxins and penicillin G confirmed the high hygienic quality of the studied product regarding fungal and mycotoxin contamination. For the first time, this study provides insights about Salame Piemonte mycobiota, which together with the bacterial microbiota and Salame Piemonte process specifications, are responsible for the product organoleptic properties.