Penicillium Cyclopium Research Articles

Assessment of lead tolerance in gamma exposed Aspergillus niger van Tieghem & Penicillium cyclopium Westling

Purpose: Present study deals with the role of gamma irradiation in modulating lead (Pb) tolerance of Aspergillus niger van Tieghem. and Penicillium cyclopium Westling.Materials and methods: After being exposed to gamma absorbed doses those fungal strains were subjected to heavy metal uptake efficacies and anti-oxidative study. Fourier Transform Infrared (FTIR) spectra and Scanning Electron Microscopic (SEM) studies were also evaluated.Result: Gamma exposed A. niger & P. cyclopium showed enhanced growth in terms of colony forming unit (CFU) and more Pb uptake efficacies compared to their un-irradiated counterparts. FTIR spectra illustrated the involvement of functional groups in Pb biosorption. SEM photographs revealed the structural deformities in both the fungal strains after being exposed to Pb and gamma. Upregulated anti-oxidative defense system (super oxide dismutase, catalase, total glutathione) in gamma exposed fungal groups are accountable for enhanced Pb tolerance and removal than that of their un-irradiated counterparts.Conclusion: The outcomes of this study exhibit a light towards a new step of heavy metal bioremediation.

Biosynthetic study of conidiation-inducing factor conidiogenone: heterologous production and cyclization mechanism of a key bifunctional diterpene synthase.

Conidiogenone, a diterpene with a unique structure, is known to induce the conidiation of Penicillium cyclopium. The biosynthetic pathway of (-)-conidiogenone has been fully elucidated by the heterologous expression of biosynthetic genes in Aspergillus oryzae and by in vitro enzyme assay with 13C-labeled substrates. After construction of deoxyconidiogenol by the action of bifunctional terpene synthase, one cytochrome P450 catalyzes two rounds of oxidation to furnish conidiogenone. Notably, similar biosynthetic genes are conserved among more than 10 Penicillium sp., suggesting that conidiogenone is a common conidiation inducer in this genus. The cyclization mechanism catalyzed by terpene synthase, which involves successive 1,2-alkyl shifts, was fully elucidated using 13C-labeled geranylgeranyl pyrophosphate (GGPP) as substrate. During the structural analysis of deoxyconidiogenol, we observed broadening of some of the 13C signals measured at room temperature, which has not been observed with other structurally related compounds. Careful examination using techniques including 13C NMR studies at -80 °C, conformational analysis and prediction of the 13C chemical shifts using density functional theory gave insights into this intriguing phenomenon.

Development of effective and safe compound disinfectant for space cabins

Space-enclosed cabins such as space stations provide a suitable living environment for astronauts, meanwhile they also facilitate various microorganisms to breed and corrode the materials' surfaces. To effectively prevent and control the microbial contamination in space-closed cabins and ensure the health of the astronauts, it is necessary to develop effective and safe disinfectants. In this study, organosilicon quaternary ammonium salt(OQAS), sodium dehydroacetate (SD) and polyhexamethylene biguanidine hydrochloride(PHMB) were selected as the target disinfectants and tested on four kinds of corrosive fungi, including Cladosporium cladosporioides, Aspergillus niger, Penicillium aurantiogriseum and Cladosporium herbarum. Firstly, the mycelial growth inhibition effects of these three disinfectants on the four fungi were tested separately, and the effective concentration range of the three disinfectants were determined. Then we combined these three disinfectants to get a new highly effective and safe compound bactericides, consisting of OQAS (0.37 mg/ml), SD (0.7 mg/ml) and PHMB (0.5 mg/ml), in which the total concentrations of bactericides were 1.57 mg/ml. This study provided a new perspective for the development of safe microbial bacteriostatic agents in spacecraft.

Penicillium cyclopium 2-11 штамындағы пектинлиаза ферменттерiнiң биосинтезiндегi фосфордың ролi

Penicillium cyclopium 2-11 штамындағы пектинлиаза ферменттерiнiң биосинтезiндегi фосфордың ролi

Effect of high temperature with Litsea cubeba Pers. to control mold growth on bamboo food packaging and its possible modes of action

This study examined the effect of high temperature and Litsea cubeba oil with its main components against molds (Aspergillus niger, Aspergillus flavus, Penicillium sp., Penicillium cyclopium, Rhizopus sp., Fusarium sp., and Cladosporium sp.) on bamboo food packaging. Response surface methodology (RSM) with X1 (concentration of L. cubeba oil at 100, 300, and 500 mg g-1), X2 (temperature at 60, 80, and 100 °C), and X3 (time at 12, 14, and 16 h) was used to find the inhibitory periods of natural mold on packaging plates. The physical properties and the change of chemical components on the bamboo packaging plate before and after temperature treatment were determined to find the mode of action using gas chromatography–mass spectrometry (GC-MS). High temperature (at 100 °C) was a good inhibitor of all mold growth with the MIC (minimum inhibitory concentration) of 300 mg g-1; without heat treatment, no MIC was found. In addition, it was found that by using 300 mg g-1 of L. cubeba oil at 100 °C with an exposure time of 12 h, spore germination on the bamboo surfaces was completely inhibited for at least 290 days. After using high temperature, citral was detected on the surface of the packaging plates. Therefore, these components could be the key factors for inhibiting molds.

Volatile compounds other than CO2 emitted by different microorganisms promote distinct posttranscriptionally regulated responses in plants.

A "box-in-box" cocultivation system was used to investigate plant responses to microbial volatile compounds (VCs) and to evaluate the contributions of organic and inorganic VCs (VOCs and VICs, respectively) to these responses. Arabidopsis plants were exposed to VCs emitted by adjacent Alternaria alternata and Penicillium aurantiogriseum cultures, with and without charcoal filtration. No VOCs were detected in the headspace of growth chambers containing fungal cultures with charcoal filters. However, these growth chambers exhibited elevated CO2 and bioactive CO and NO headspace concentrations. Independently of charcoal filtration, VCs from both fungal phytopathogens promoted growth and distinct developmental changes. Plants cultured at CO2 levels observed in growth boxes containing fungal cultures were identical to those cultured at ambient CO2 . Plants exposed to charcoal-filtered fungal VCs, nonfiltered VCs, or superelevated CO2 levels exhibited transcriptional changes resembling those induced by increased irradiance. Thus, in the "box-in-box" system, (a) fungal VICs other than CO2 and/or VOCs not detected by our analytical systems strongly influence the plants' responses to fungal VCs, (b) different microorganisms release VCs with distinct action potentials, (c) transcriptional changes in VC-exposed plants are mainly due to enhanced photosynthesis signaling, and (d) regulation of some plant responses to fungal VCs is primarily posttranscriptional.

Mycoviruses mediate mycotoxin regulation in Aspergillus ochraceus.

To date, no demonstration of a direct correlation between the presence of mycoviruses and the quantitative or qualitative modulation of mycotoxins has been shown. In our study, we transfected a virus-free ochratoxin A (OTA)-producing isolate of Aspergillus ochraceus with purified mycoviruses from a different A. ochraceus isolate and from Penicillium aurantiogriseum. Among the mycoviruses tested, only Aspergillus ochraceus virus (AoV), a partitivirus widespread in A. ochraceus, caused a specific interaction that led to an overproduction of OTA, which is regulated by the European Commission and is the second most important contaminant of food and feed commodities. Gene expression analysis failed to reveal a specific viral upregulation of the mRNA of genes considered to play a role in the OTA biosynthetic pathway. Furthermore, AoOTApks1, a polyketide synthase gene considered essential for OTA production, is surprisingly absent in the genome of our OTA-producing isolate. The possible biological and evolutionary implications of the mycoviral regulation of mycotoxin production are discussed.

MORPHOLOGICAL IDENTITY AND POPULATION STRUCTURE OF HEMIBIOTROPHIC FUNGUS Colletotrichum coccodes COLONIZING PEPPER PLANTS

Colletotrichum coccodes (Wallr.) Hughes has been recognized as one of casual agents of anthracnose roots pepper in south-eastern Poland. During 2007–2012 the species was isolated from roots of sweet pepper (Capsicum annuum L.) cultivated in the field. The purpose of the study was morphological characterization and biotic activity of C. coccodes isolates. Five randomly chosen isolates from fungus population and one reference isolate obtained from Bank of Plant Pathogens and Investigation of their Biodiversity of IPP-NRI in Poznan in Poland were studied. The character of culture like growth rate, the colour of averse and reverse and the formation of morphological structures of the fungus such as acervuli, conidia, sclerotia were studied. Ultrastructural observations of morphological structures were made using light and scanning electron microscopy. Biotic activity of C. coccodes was conducted using the method of biotic series on PDA. Seven species of test fungi were used in the study: Alternaria alternata, Aureobasidium pullulans, Gibberella avenacea, G. intricans, Fusarium oxysporum, Penicillium aurantiogriseum, Trichoderma harzianum. The biotic activity test showed that C. coccodes is a weak competitor, and its development in the rhizosphere of sweet pepper may be limited by numerous antagonists.

Skład chemiczny etanolowego ekstraktu z propolisu i jego aktywność biologiczna wobec grzybów pleśniowych

Introduction. Propolis is a natural material whose extracts indicate numerous biological activities, including antifungal, antibacterial, antioxidant and anticancer. Phenolic compounds (flavonoids as well as phenolic acids and their esters) are one of the most frequently mentioned group of propolis components responsible for the biological activity of propolis extracts. Another group of components identified in propolis is elements, including macro- and microelements as well as heavy metals. Aim. The aim of the study was to determine the biological activity of the ethanolic propolis extract originated form Warmian-Masurian Voivodeship against moulds and to analyze concentration of selected flavonoids and elements present in the extract. Material and methods. The biological activity of the propolis extract obtained from a raw propolis originated from Warmian-Masurian Voivodeship against moulds was determined by dilution in nutrient solution. The content of selected flavonoids (apigenin, pinocembrin, kaempferol and galangin) in the propolis extract was determined using ultra-performance liquid chromatography photodiode array detection tandem mass spectrometric method. The concentration of elements in the propolis extract was analyzed by flame atomic absorption spectrometry. Results. The propolis extract was characterized by activity against all tested fungal strains. It showed the highest activity towards Chaetomium globosum and Penicillium cyclopium. Among analyzed flavonoids, the highest concentration in the propolis extract was determined for pinocembrin. In addition, high concentrations of Na, Mg, K, Ca and Fe were found in the propolis extract, and no toxic heavy metals were found. Conclusions. The ethanolic propolis extract of national origin shows biological activity against tested moulds and it is a valuable source of flavonoids, as well as macro- and microelements.

Resistance of fungal growth on Scots pine treated with caffeine

Alkaloids like caffeine are well-known compounds of natural origin, economically suitable and commercial available, which could facilitate their future use for wood protection. The aim of our study was to determine the resistance of Scots pine (Pinus sylvestris L.) treated with various concentrations of caffeine water solutions (25–0.4 mg mL−1) against common mold fungi and four wood-decaying fungi (Coniophora puteana, Poria placenta, Gloeophyllum trabeum, and Trametes versicolor). Half of the treated specimens before fungal exposition were subjected to leaching procedure. Mass loss from brown rotting fungi was significantly inhibited by all preparations. Moreover, caffeine imparted high resistance in Scots pine to the white-rot fungus T. versicolor. In this study we have shown that to effectively protect wood against all tested decay fungi the concentration of the caffeine solution should be 10 mg mL−1. Complete inhibition of the growth of Aspergillus niger, A. terreus, Chaetomium globosum, Cladosporium herbarum, Paecilomyces variotii, Penicillium cyclopium, P. funiculosum, Trichoderma viride, except A. versicolor and Phoma violacea has been achieved by applying a higher concentration of 25 mg mL−1 of caffeine. Mold and decay resistance of wood treated caffeine is probably related to fact that caffeine has been shown to inhibit chitinases activity, which results in inhibition of fungal growth.

Short communication: Cheese supplemented with Thymus algeriensis oil, a potential natural food preservative

The essential oil of Thymus algeriensis was analyzed as a potential preservative in soft cheese. We developed a novel method to test the preserving properties of essential oil in soft cheese. Contamination incidence of Penicillium aurantiogriseum was absent after 30 d of storage at 4°C with 25 µL of essential oil added. The antimicrobial activity was tested against 8 bacteria and 8 fungi. Thymus algeriensis oil showed inhibitory activity against tested bacteria at 0.03 to 0.09 mg/mL, and bactericidal activity was achieved at 0.05 to 0.15 mg/mL. For antifungal activity, minimum inhibitory concentrations ranged between 0.01 and 0.04 mg/mL and minimum fungicidal concentrations between 0.01 and 0.04 mg/mL. Furthermore, the oil was also screened for antiradical activity using the 1,1-diphenyl-2-picrylhydrazyl assay. The results showed that the oil was active and achieved half-maximal inhibitory activity at 0.132 mg/mL. We used gas chromatography, gas chromatography-mass spectrometry, and nuclear magnetic resonance spectrometry to investigate the volatile compounds from the oil. Carvacrol was identified as the main compound in the oil, represented by 80.9% of the total constituents, followed by p-cymene (7.7%).

Single step purification via magnetic nanoparticles of new broad pH active protease from Penicillium aurantiogriseum

A new set of applications can be achieved when using high stability proteases. Industrially, high costs can be related to production medium and purification process. Magnetic nanoparticles have been successfully used for rapid and scalable purification. In this work, azocasein were immobilized on magnetite nanoparticles and applied in a single step purification of protease produced by Penicillium aurantiogriseum using soybean flour medium, and the new purified enzyme was characterized. Glutaraldehyde activated nanoparticles were used in azocasein immobilization and then incubated with dialyzed 60–80% saline precipitation fraction of crude extract for purification. Adsorbents were washed 7 times (0.1 M NaCl solution) and eluted 3 times (1 M NaCl solution), these final elutions contained the purified protease. This protease was purified 55.68-fold, retaining 46% of its original activity. Presented approximately 40 kDa on SDS-PAGE and optimum activity at 45 °C and pH 9.0. Maintained over 60% of activity from pH 6.0 to 11.0. Kept more than 50% activity from 15 to 55 °C, did not lose any activity over 48 h at 25 °C. Inhibitors assay suggested a serine protease with aspartic residues on its active site. Results report a successful application of an alternative purification method and novel broad pH tolerant protease.

Invertase Production by Fungi, Characterization of Enzyme Activity and Kinetic Parameters

The present paper presents the results of research carried out on 14 fungal isolates of various origins aiming to select new efficient sources for invertase production for further biotechnological application. Aspergillus flavus presented the highest protein content and Aspergillus niger, the most intense invertase activity. The relationship between enzyme concentration and enzymatic activity at 0.25 mM mL-1sucrose as substrate assayed for successive decimal dilutions of Aspergillus niger enzyme ranging from 0.1 to 1mlLrevealed a linear correspondence between 0.1 and 0.5mL. The kinetic parameters Michaelis-Menten constant (Km) and maximal velocity (Vmax) for invertase activity of Aspergillus niger, Penicillium aurantiogriseum, Aspergillus wentii and Rhizopus stolonifer were calculated. The determination coefficients R2 calculated from Lineweaver-Burk plots presented values very close or equal to 1.

Transmission of Penicillium aurantiogriseum partiti-like virus 1 to a new fungal host (Cryphonectria parasitica) confers higher resistance to salinity and reveals adaptive genomic changes.

We attempted to transfect six recently characterized virus species to protoplasts of Penicillium janczewskii and Chryphonectria parasitica. None of the recovered P. janczewskii colonies was positive for the transfected viruses, but Penicillium aurantiogriseum partiti-like virus 1 (PaPLV1) was detected in three distinct regenerated C. parasitica colonies. We screened the phenotype of the infected strains in up to 45 different conditions combining different media, salinity and temperatures: our results show that the infected strains grow slower than the virus- free in most of the tested conditions with the exception of halophilic stress in a specific nutrient combination media. We proceeded to characterize molecularly the population of distinct isolates of PaPLV1 infected C. parasitica through RNAseq: comparison to the viral population present in the original host - P. auratiogriseum - showed that two isolates accumulated non-synonymous mutations suggesting adaptation to the new host. RNAseq analyses identified a second genomic RNA segment and northern blot of RNA extracted from purified virus suspensions allowed establishing that PaPLV1 is at least bipartite in nature and that it forms isometric virions of circa 36-38nm in diameter. In light of these new acquisitions, we discuss the taxonomic placement of PaPLV1 inside the Partitiviridae.

Optimization of Penicillium aurantiogriseum protease immobilization on magnetic nanoparticles for antioxidant peptides’ obtainment

ABSTRACTThis work reports an optimization of protease from Penicillium aurantiogriseum immobilization on polyaniline-coated magnetic nanoparticles for antioxidant peptides’ obtainment derived from bovine casein. Immobilization process was optimized using a full two-level factorial design (24) followed by a response surface methodology. Using the derivative, casein was hydrolyzed uncovering its peptides that were sequenced and had antioxidant properties tested through (2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) (ABTS) radical scavenging and hydrogen peroxide scavenging assays. Optimal conditions for immobilization were 2 hr of immobilization, offered protein amount of 200 µg/mL, immobilization pH of 6.3 and 7.3 hr of activation. Derivative keeps over 74% of its original activity after reused five times. Free and immobilized enzyme casein hydrolysates presented similar peptide mass fingerprints, and prevalent peptides could be sequenced. Hydrolysates presented more than 2.5× higher ROS scavenging activity than nonhydrolyzed casein, which validates the immobilized protease capacity to develop casein-derived natural ingredients with potential for functional foods.

Mycoviruses of an endophytic fungus can replicate in plant cells: evolutionary implications

So far there is no record of a specific virus able to infect both fungal and plant hosts in nature. However, experimental evidence shows that some plant virus RdRPs are able to perform replication in trans of genomic or DI RNAs in the yeast Saccharomyces cerevisiae. Furthermore, tobacco mosaic virus was recently shown to replicate in a filamentous ascomycetous fungus. Thus, at least experimentally, some plant viruses can infect some fungi. Endophytic fungi have been reported from many plants and several of these fungi have been shown to contain viruses. Here we tested if mycoviruses derived from a marine plant endophyte can replicate in plant cells. For this purpose, we used partially purified viral particles from isolate MUT4330 of Penicillium aurantiogriseum var. viridicatum which harbors six virus species, some having dsRNA and some positive-strand ssRNA genomes. These were transfected into three distinct plant protoplast cell systems. Time-course analysis of absolute RNA accumulation provided for the first time evidence that viruses of two species belonging to the Partitiviridae and Totiviridae families, can replicate in plant cells without evidence of host adaptation, i.e, changes in their nucleotide sequence.

Raw and waste plant materials as sources of fungi with epoxide hydrolase activity. Application to the kinetic resolution of aryl and alkyl glycidyl ethers

The by-products of olive oil production can be used as sources of microbial strains. Penicillium sp., Aspergillus terreus, Penicillium aurantiogriseum, Aspergillus tubingensis and Aspergillus niger were selected on the basis of their epoxide-hydrolyzing activity towards racemic rac-glycidyl phenyl ether. We studied the effect on enzymatic activity of adding styrene oxide to the growth medium. It induced the biosynthesis of epoxide hydrolases and reduced cell growth. The resolution capacity of the five fungi was tested on rac-glycidyl phenyl ether, rac-benzyl glycidyl ether, rac-1,2-epoxyhexane and rac-1,2-epoxyoctane. The resolution of rac-glycidyl phenyl ether by A. niger, rac-benzyl glycidyl ether by P. aurantiogriseum and A. terreus, rac-1,2-epoxyhexane by A. tubingensis and rac-1,2-epoxyoctane by A. terreus provided (S)-3-phenoxy-1,2-propanediol (45.1% yield, 51.4% ee), (R)-3-benzyloxy-1,2-propanediol (40.8% yield, 43.3% ee), (S)-3-benzyloxy-1,2-propanediol (45.4% yield, 45.6% ee), (R)-1,2-hexanediol (70.4% yield, 24.4% ee) and (R)-1,2-octanediol (21.4% yield, 27.5% ee), respectively. The (R)-enantiopreference of the epoxide hydrolases from P. aurantiogriseum is unprecedented.

Anti-fungal activity, mechanism studies on \u03b1-Phellandrene and Nonanal against Penicillium cyclopium

BackgroundEssential oils from plants have been reported to have wide spread antimicrobial activity against various bacterial and fungal pathogens, and these include α-Phellandrene, Nonanal and other volatile substances. However, biological activities of α-Phellandrene and Nonanal have been reported only in a few publications. Further investigations are necessary to determine the antimicrobial activity of these compounds, especially for individual application, to establish the possible mechanism of action of the most active compound.ResultsThe results are shown that α-Phellandrene and Nonanal have a dose-dependent inhibition on the mycelial growth of Penicillium cyclopium. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) are 1.7 and 1.8 mL/L for α-Phellandrene, 0.3 and 0.4 mL/L for Nonanal, respectively. The volatile compounds altered the morphology of P. cyclopium hyphae by causing loss of cytoplasmic material and distortion of the mycelia. The membrane permeability of P. cyclopium increased with increasing concentrations of the two volatile compounds, as evidenced by cell constituent release, extracellular conductivity and induced efflux of K+. Moreover, the two volatile compounds induced a decrease in pH and in the total lipid content of P. cyclopium, which suggested that cell membrane integrity had been compromised.ConclusionsThe results demonstrated that α-Phellandrene and Nonanal could significantly inhibit the mycelia growth of P. cyclopium by severely disrupting the integrity of the fungal cell membrane, leading to the leakage of cell constituents and potassium ions, and triggering an increase of the total lipid content, extracellular pH and membrane permeability. Our present study suggests that α-Phellandrene and Nonanal might be a biological fungicide for the control of P. cyclopium in postharvest tomato fruits.

Improvement in thermostability of an alkaline lipase I from Penicillium cyclopium by directed evolution

A novel alkaline-stable lipase I from Penicillium cyclopium with improved thermostability was prepared by molecular modification.

Statistical optimization of L-asparaginase production by using Fusarium solani

Background and objective The enzyme l-asparaginase is important as a therapeutic agent in the treatment of acute lymphocytic leukemia. It has been observed that microorganisms such as yeast and filamentous fungi such as Aspergillus spp., Penicillium spp., and Fusarium spp. are commonly produced asparaginases with fewer adverse effects. Fusarium solani was selected to be the most potent microbial isolate for l-asparaginase production. The factors controlling l-asparaginase production, such as containing media, were implemented to increase the yield of l-asparaginase. Statistical experimental design such as the Plackett–Burman enables finding out the most effective factors that increase the yield of l-asparaginase. Materials and methods Aspergillus rubber, Aspergillus terreus, Epico niger, Penicillium cyclopium, and F. solani were chosen for screening the l-asparaginase activity using phenol red – a pH indicator. l-Asparagine and the modified Czapek Dox media were tested for the production of l-asparaginase from F. solani under different incubation times. To optimize l-asparaginase production by using F. solani, the combined effect of seven variables (l-asparagine, sucrose, NaNo3, K2HPO4, MgSO4, KCl, and time) were assessed using the central composite design. Results and conclusion All the four fungal strains screened for l-asparaginase activity showed positive results in the rapid plate assay method. The medium employed contained asparagine with phenol red, and after incubation, pink zones around the colonies were observed. F. solani was a potential source for a high yield of l-asparaginase enzyme and high substrate specificity. Modified Czapek medium II was the most suitable for l-asparaginase, showing an activity of 121 U/ml. The result on l-asparaginase activity according to the screening Plackett–Burman experiments gives a medium composed of the following (g/l): sucrose (1); l-asparagine (0.5); KH2PO4. (0.75), MgSO4 (0.7); H2O (0.72); KCl (0.72); (NH4)2SO4 (11.7) at an incubation time of 3 days. For further investigation, x1 K2HPO4, x2 sucrose, and x3 time were found to be the most significant variables affecting l-asparaginase activity. The second optimization step was to identify optimal values of the three factors that bring about maximum l-asparaginase activity, using the central composite designed experiment. The results showed that K2HPO4 (2.5 g/l), sucrose (4 g/l), and the time (8 days) were critical in the production of l-asparaginase.