Collagenolytic activity in several species of deuteromycetes under various storage conditions

The ability of deuteromycetes of the genera Penicillium, Aspergillus, and Botrytis to retain collagenolytic activity was studied after both 2 and 10 years of storage on a Czapek medium under a layer of mineral oil at 4°C, as well as in silica gel granules at 20 and ?60°C. The enzymatic activity of several species, including Botrytis terrestris, Penicillium janthinellum, Penicillium chrysogenum, and Penicillium citrinum, was retained under both conditions of storage. Aspergillus repens retained enzymatic activity only if stored under a layer of mineral oil. The viability of conidia and the collagenolytic activity of Botrytis terrestris, P. janthinellum, P. chrysogenum, and Penicillium citrinum, maintained on silica gel for 10 years, depended on the storage temperature. The viability of the test strains improved after storage on a silica gel at ?60°C. A strain of Aspergillus repens lost its ability to dissolve collagen at various storage tempeatures on the silica gel. The index of lysis for three strains of Penicillium deuteromycetes (Penicillium janthinellum, Penicillium chrysogenum, and Penicillium citrinum) increased after a 10-year storage on silica gel at ?60°C.     

Hydrolysis of PET and bis-(benzoyloxyethyl) terephthalate with a new polyesterase from Penicillium citrinum

A polyethylene terephthalate (PET) model substrate, bis-(benzoyloxyethyl)terephthalate (3PET), was used to screen for micro-organisms producing enzymes hydrolyzing PET. From this screen, a strain growing on 3PET was isolated and identified as Penicillium citrinum. The polyesterase responsible for 3PET and PET hydrolysis was purified to electrophoretic homogeneity. The polyesterase had a molecular weight of 14.1 kDa, and the K_m and K_cat values on 4-nitrophenyl butyrate were 0.57?mM and 0.21?s^?1, respectively. Highest enzyme activities were obtained when P. citrinum was grown on a medium containing cutin, which was hydrolyzed by the polyesterase. Surface hydrolysis of PET with the enzyme lead to an increase in hydrophilicity based on rising height (+5.1?cm) and drop dissipation measurements (55?s). Both from PET and 3PET bis-(2-hydroxyethyl)terephthalate and mono-(2-hydroxyethyl)terephthalate were released, while only low amounts of terephthalic acid were liberated.     

Use of the Weibull model to describe inactivation of dry‐harvested conidia of different Penicillium species by ethanol vapours

Aims: This study aimed at modelling the effect of ethanol vapours, in the range 0·7–7·5 kPa, on the inactivation of dry‐harvested conidia of Penicillium chrysogenum, Penicillium digitatum and Penicillium italicum. Methods and Results: Survival curves were modelled by a Weibull model: log (N/N₀) = ?1/2·303 (t/α)^β. The shape parameter β was different from one in all cases, indicating that the classical first‐order kinetics approach is the exception rather than the rule. Survival curves exhibited upward concavity (β < 1) with the notable exception of P. chrysogenum at ethanol vapour pressures 0·7 and 1·5 kPa. The scale parameter α (h) varied greatly depending on the ethanol vapour pressure and on the species. Conclusions: For safety reasons, it is recommended not to exceed an ethanol vapour pressure of 3·3 kPa. At 2·8 kPa, more than 4 log₁₀ reductions in viable conidia were achieved for all the species after 24‐h exposure. Significance and Impact of the Study: Ethanol has GRAS status in the USA and represents an interesting alternative to fungicides. The effectiveness of ethanol vapours to inactivate dry‐harvested conidia of some Penicillium was demonstrated in this study.     

Fungal rock phosphate solubilization using sugarcane bagasse

The effects of different doses of rock phosphate (RP), sucrose, and (NH₄)₂SO₄ on the solubilization of RP from Araxá and Catal?o (Brazil) by Aspergillus niger, Penicillium canescens, Eupenicillium ludwigii, and Penicillium islandicum were evaluated in a solid-state fermentation (SSF) system with sugarcane bagasse. The factors evaluated were combined following a 2³?+?1 factorial design to determine their optimum concentrations. The fitted response surfaces showed that higher doses of RP promoted higher phosphorus (P) solubilization. The addition of sucrose did not have effects on P solubilization in most treatments due to the presence of soluble sugars in the bagasse. Except for A. niger, all the fungi required high (NH₄)₂SO₄ doses to achieve the highest level of P solubilization. Inversely, addition of (NH₄)₂SO₄ was inhibitory to P solubilization by A. niger. Among the fungi tested, A. niger stood out, showing the highest solubilization capacity and for not requiring sucrose or (NH₄)₂SO₄ supplementation. An additional experiment with A. niger showed that the content of soluble P can be increased by adding higher RP doses in the medium. However, P yield decreases with increasing RP doses. In this experiment, the maximal P yield (approximately 60?%) was achieved with the lower RP dose (3?g?L^?1). Our results show that SSF can be used to obtain a low cost biofertilizer rich in P combining RP, sugarcane bagasse, and A. niger. Moreover, sugarcane bagasse is a suitable substrate for SSF aiming at RP solubilization, since this residue can supply the C and N necessary for the metabolism of A. niger within a range that favors RP solubilization.     

Control of green mould of citrus by using Trichoderma harzianum,humic acid or garlic

Penicillium digitatum, an aggressive fungus causes post-harvest decay of mandarin sweet orange and Washington navel. In vitro Trichoderma harzianum or humic acid (HA) or powdered cloves of garlic caused inhibition of fungal growth of isolates P₁ and P₂. Under storage conditions, the fruit citrus is protected by using T. harzianum with standard volume 2.0?ml (9.6?×?10⁶?conidia/ml) and application 24?h before inoculation reduces disease incidence and disease severity after seven?days from inoculation with P. digitatum spore suspension (1.0?×?10⁶?spores/ml) compared to control. Spraying the fruit citrus by standard volume of 2.0?ml of either HA or powder cloves of garlic 1% on each fruit 24?h before inoculation reduces disease incidence and disease severity after seven?days from inoculation with P. digitatum (1.0?×?10⁶ spores/ml) compared to control. The lowest percentage of disease incidence and disease severity were associated with powder of cloves garlic and followed by HA and T. harzianum during two growing seasons compared with the untreated and control.     

Penicillium strains isolated from Slovak grape berries taxonomy assessment by secondary metabolite profile

The secondary metabolite profiles of microfungi of the genus Penicillium isolated from samples of grape berries collected in two different phases during two vegetative seasons in Slovakia is described to assess the taxonomy. Three Slovak vine regions have been selected for this study, based on their climatic differences and national economic importance. Cultures of microfungi isolated from berries were incubated on different selective media for macro and micromorphology identification. The species Penicillium brevicompactum, Penicillium crustosum, Penicillium chrysogenum, Penicillium expansum, Penicillium palitans and Penicillium polonicum were identified according to growth and morphology. The related strains were found to produce a broad spectrum of fungal metabolites, including roquefortine C, chaetoglobosin A, penitrem A, cyclopeptin, cyclopenin, viridicatin, methylviridicatin, verrucofortine, secalonic acid D, cyclopiazonic acid, fumigaclavine and mycophenolic acid. Chemotaxonomy was performed using high-performance liquid chromatography (HPLC) and mass spectrometry (MS). Dried grape berries were also analyzed allowing to assess the presence of patulin, roquefortine C and penicillic acid; this last one has been identified in dried berries but not in vitro.     

Vanadate inhibits the production and/or release of secondary metabolites without impairing growth in several fungal species

Vanadate (NaVO₃) in concentrations between 0.1–3.0 mmol/L inhibited the production of secondary metabolites (SMs) of strains of the following species:Trichoderma viride, Penicillium purpurogenum, Penicillium citrinum, Talaromyces avellaneus, andVerticillium psalliotœ. Growth was either not affected by NaVO₃, or the inhibition of the SM production occurred at lower NaVO₃, concentrations than that of the growth. Thus, at some NaVO₃ concentration the SM production was inhibited but the growth remained unaffected. The results suggest that NaVO₃ exerts a specific action either on the SM biosynthetic pathway(s) or on the export of SMs from cells.     

Oxidation of Polyamines by Fungal Enzymes

Polyamine oxidase was found in mycelia of fungi belonging to the genera of Aspergillus, Mucor, Penicillium, Rhizopus, Cylindrocarpon, Fusarium and Gibberella when they were grown in medium containing spermine or spermidine as the sole source of nitrogen. The maximal formation of the enzymes of Penicillium chrysogenum and Aspergillus terreus was observed in early stationary phase of growth, and thereafter, the enzymes disappeared with consumption of substrate. The oxidation products of spermine and spermidine by the two enzymes were identified as putrescine, 3-aminopropionaldehyde and H₂O₂. Therefore, the enzymes were characterized as a type of polyamine oxidase of rat liver.     

Purification of a Penicillium citrinum lipase by chromatographic processes

A lipase from a wild strain of Penicillium citrinum was purified by ammonium sulphate precipitation, gel filtration chromatography on a Superose 6 column and hydrophobic interaction chromatography (HIC) on a Phenyl Superose column. The yield and purification factor were 15.2% and 379 fold, respectively. The gel filtration step was efficiently scaled-up in a Superose 6 preparative grade column and after this step, the lipase was recovered in the form of a high molecular weight aggregate. The partial disaggregation of the complex was achieved by HIC and elution with 1.0% (w/v) CHAPS. The lipase produced by Penicillium citrinum forms a dimmer of 63?000 Da, as determined by SDS-PAGE, and it accumulates in the fermentation broth as high molecular weight aggregates (>2?00?000 Da). The analysis of the dimmer showed two subunits with similar molecular weights (31?000–33?000 Da) and isoelectric points (4.8–5.0).     

Isolation,Identification and Screening of Manganese Solubilizing Fungi From Low-Grade Manganese Ore Deposits

The present investigation reports the isolation, molecular identification and screening of manganese (Mn) solubilizing fungal strains from low-grade Mn mine tailings. Six morphologically distinct Mn solubilizing fungal strains were isolated on MnO₂-supplemented agar plates with Mn concentration of 0.1% (w/v). The biochemical characterization of the isolated fungal strains was carried out. The molecular identification by internal transcribed spacer (ITS) sequencing identified the strains as Aspergillus terreus, Aspergillus oryzae, Penicillium sp., Penicillium sp., Penicillium daleae and Penicillium sp. with GenBank accession numbers KP309809, KP309810, KP309811, KP309812, KP309813 and KP309814, respectively. The ability of the isolated fungal strains to tolerate and solubilize Mn was investigated by subculturing them on Mn-supplemented plates with concentration ranging from 0.1 to 0.5% (w/v). Mn solubilizing ability of the fungal isolates is possibly due to the mycelia production of biogenerated organic acids such as oxalic acid, citric acid, maleic acid and gluconic acid as revealed by ion chromatography. Our investigation signifies the role of fungi in biotransformation of insoluble Mn oxide.     

The cellulolytic activity of the culture filtrates of egyptian mould fungi

Summary The cellulolytic activity of the culture filtrates of two strong cellulose decomposers namely Penicillium oxalicum and Helminthosporium cyclops was studied. The culture age influenced markedly the cellulolytic activity and two weeks of growth were found to establish the highest activity. The highest activity was recorded at p_H 4 and p_H 5 in case of Penicillium oxalicum and Helminthosporium cyclops respectively. In both experimental fungi, 40° C was the most suitable temperature for the cellulolytic action. The cellulase activity of the filtrate was found to be comparatively stable for a long time. The cellulase enzymes of both organisms were found to be strongly adaptive.     

Production and characterization of Penicillium brasilianum pectinases with regard to industrial application

The objective of this study was to evaluate the production of pectinase by an isolated strain of Penicillium brasilianum in a bioreactor and to consider its potential for industrial applications (i.e. fruit juice). The optimization of production was achieved through experimental design. The maximum exo-polygalacturonase (Exo-PG) production in the bioreactor was 53.8?U mL^?1 under the conditions of 180?rpm, an aeration rate of 1.5 vvm, 30?°C, pH_initial of 5.5, 5?×?10⁶ spores mL^?1, 32?g L^?1 pectin, 10?g L^?1 of yeast extract and 0.5?g L^?1 magnesium sulfate and bioproduction for 36?h. The production of Exo-PG in the bioreactor was 1.3 times higher than that obtained in shake flasks, with aeration (1.5 vvm) and agitation (180?rpm) control. The crude enzyme complex, beyond the pectinolytic activity of Exo-PG (53.8?U mL^?1), also contained activity pectin methylesterase (6.0?U mL^?1) and pectin lyase (6.61?U mL^?1). At a crude enzyme complex with a concentration of 0.5% (v/v), viscosity of peach juice was reduced by 11.66%, turbidity was reduced by 13.71% and clarification was increased by 26.92%. Based on the present results, we can conclude that the new strain of isolated P. brasilianum produced high amounts of pectinases in a bioreactor with mechanical agitation, and has the potential to be applied to in the clarification of juices.     

Purification and partial characterization of exopolygalacturonase I from Penicillium frequentans

A polygalacturonase with a molecular mass of 74 kDa, an isoelectric point around pH 4.2 and pH – and temperature optima of 3.9 and 50°C, respectively, was purified from a culture fluid of Penicillium frequentans. The enzyme was characterized as an exo-α-1,4-polygalacturonase (exo-PG I). K_m and V_max for sodium polypectate hydrolysis were 0.68 g/l and 596.8 U × mg⁻¹, respectively. The enzyme, a glycoprotein with a carbohydrate content of 81%, is probably the main pectinase of Penicillium frequentans responsible for cleaving monomer units from the non-reducing end of pectin.     

Nutritional requirements of antagonists to peach twig blight,Monilinia laxa,in relation to biocontrol

Different carbon and nitrogen sources and accessory substances were tested to determine their effect on the growth and sporulation of the peach twig blight pathogen,Monilinia laxa, and of three of its antagonists (Penicillium frequentans, Penicillium purpurogenum andEpicoccum nigrum), since the success in twig blight biological control by treatments with the fungal antagonists depends on the type of nutrients added to the antagonist formulation. Combinations of sucrose-ammonium tartrate, glucose-(NH₄)₃PO₄-folic acid and lactose-KNO₃ were selected from these laboratory experiments because they enhanced the growth and sporulation ofP. frequentans, P. purpurogenum andE. nigrum, respectively, but notM. laxa. In glasshouse experiments, twig blight was reduced following the application of mixtures of antagonists with the corresponding enhancing nutrients.     

Studies on the ability of mycotoxins to diffuse in bread

Summary The moulds Aspergillus parasiticus (aflatoxins B₁, B₂, G₁, G₂, and M₁), A. ochraceus (ochratoxin A) and Penicillium chrysogenum (citrinin) were grown on whole wheat bread either in the presence or absence of oxygen. In the presence of oxygen, both A. parasiticus and A. ochraceus developed dense colonies and formed considerable amounts of mycotoxins whereas Penicillium chrysogenum only grew and produced citrinin on the surface of the bread. In the absence of oxygen fungal growth did not occur and most of the toxins were undetectable even in regions of bread immediately adjacent to the moulds although a very slight diffusion of the aflatoxins B₁ and G₁ through 1 cm was observed. It is concluded that diffusion of the tested mycotoxins from hyphae into bread is not a problem for food safety.     

Investigation of submerged production of the antibiotic cyanein

В ходе глубинной ферментации Penicillium cyaneum исследовали зависимость выхода цианеина от состава ферментационной среды. Было установлено, что биосинтез антибиотика осуществляется и в простой синтетической среде с прибавлением микроэлэментов. При использовании сахарозы в качестве источника C, а KNO₃—в качестве источника N максимальная продукция 230 μг/мл была получена у неселекционированного штамма S-11 через 168 час.     

Isolation and Molecular Identification of Fungi with Magnesite Enrichment Potential from KÜMAŞ Quarries in Turkey

Abstract

Magnesite is an important raw material used in various industrial applications, especially the production of high-temperature resistant materials. Due to its high reactant nature, magnesite ore is not found in pure form and it contains a great variety of pollutants such as calcium compounds, which restrict its use when exceeding 1% of the ore. Thus, the development of efficient strategies for the removal of pollutants remains a crucial step for magnesite utilization. In this regard, our present work was conducted to isolate and identify active fungal strains that remove calcium pollutants without changing the main magnesium content of the ore. For this aim, magnesite ore samples were collected from two quarries (Turanoca?? and Ortaocak) of KÜMA? Magnesite Inc. and fungal isolation studies were done by using the ore’s flora. Active isolates were chosen according to their CaCO₃ and MgCO₃ dissolving capabilities and identified by using conventional light microscopy and molecular characterization techniques. 71 fungal isolates were obtained from the isolation step and 14 of them were chosen as active isolates that solve calcium compounds while not affecting the magnesium component. The data of the microscopic examination and 18S rDNA gene sequence analysis showed that 14 active strains with magnesite enrichment potential grouped in Aspergillus alliaceus (3), Aspergillus flavus (2), Aspergillus leporis (1), Aspergillus nomius (1), Fusarium tricinctum (2), Penicillium chrysogenum (1) and Penicillium sp. (4).     

A New Enzyme,Agmatine Oxidase,from Fungi

A new enzyme, agmatine oxidase, was found in Penicillium chrysogenum. The oxidation products of agmatine with the enzyme were identified as γ-guanidinobutyraldehyde, NH₃ and H₂O₂. The enzyme rapidly oxidized agmatine, and slightly oxidized histamine, putrescine, 1,3-diaminopropane and cadaverine. Monoamines, polyamines and guanidyl derivatives were not oxidized by the enzyme. Maximal formation of the enzyme of P. chrysogenum was observed in the early stationary phase of growth, and thereafter the enzyme disappeared with consumption of substrate. In addition to agmatine, spermine, spermidine and putrescine were also effective as nitrogen sources. Agmatine oxidase was found in mycelia of fungi belonging to the genera of Aspergillus, Penicillium, Absidia, Fusarium, Mucor, Gibberella, Cylindrocarpon and Monascus when they were grown in agmatine-containing medium.     

Phytochemical evaluation and in vitro antifungal activities of Melaleuca styphelioides leaves: comparison between volatile and non-volatile extracts

Abstract

Melaleuca styphelioides is considered as medicinal plant. This study was carried out to evaluate for the first time the phytochemical composition and to compare the antifungal activities of essential oil (EOs), methanol and aqueous extracts of M. styphelioides Sm. leaves against three fungi (Aspergillus niger, Rhizopus nigricans and Penicillium digitatum). A total of 10 components of the EO were identified, with the principal compound being methyl eugenol (87.2%). Results of the phytochemical analysis of leaves extract exhibited the presence of different phytoconstituents (phenolic compounds, flavonoids, tannins and anthocyanins). Volatile and non-volatile extracts were found to express dose-dependent inhibition against all tested fungi. Indeed, the EO oil showed significant inhibition of fungal growth and the IC₅₀ was 2.08?µL/mL for A. niger indicating that M. styphelioides leaf EO was particularly effective against this pathogen. The most susceptible species for the aqueous extract was P. digitatum (IC₅₀= 9.54?mg/mL) whereas R. nigricans was found to be more susceptible to the methanolic extract (IC₅₀= 8.31?mg/mL). Thus, the EO and aqueous as well as methanol extracts of M. styphelioides leaves possess antifungal activity and hence, it can be suggested for use in the food or pharmaceutical industries as an alternative to chemical preservatives.     

COMPARATIVE STUDIES ON RESPIRATION : XII. A COMPARISON OF THE PRODUCTION OF CARBON DIOXIDE BY PENICILLIUM AND BY A SOLUTION OF DEXTROSE AND HYDROGEN PEROXIDE.

A neutral solution of dextrose and hydrogen peroxide acts like Penicillium chrysogenum in producing an increased amount of CO₂ upon the addition of acid, but not upon the addition of alkali.