Antimicrobial activity and acetylcholinesterase inhibition by extracts from chromatin modulated fungi

ABSTRACT Major health challenges as the increasing number of cases of infections by antibiotic multiresistant microorganisms and cases of Alzheimer's disease have led to searching new control drugs. The present study aims to verify a new way of obtaining bioactive extracts from filamentous fungi with potential antimicrobial and acetylcholinesterase inhibitory activities, using epigenetic modulation to promote the expression of genes commonly silenced. For such finality, five filamentous fungal species (Talaromyces funiculosus, Talaromyces islandicus, Talaromyces minioluteus, Talaromyces pinophilus, Penicillium janthinellum) were grown or not with DNA methyltransferases inhibitors (procainamide or hydralazine) and/or a histone deacetylase inhibitor (suberohydroxamic acid). Extracts from T. islandicus cultured or not with hydralazine inhibited Listeria monocytogenes growth in 57.66 ± 5.98% and 15.38 ± 1.99%, respectively. Increment in inhibition of acetylcholinesterase activity was observed for the extract from P. janthinellum grown with procainamide (100%), when compared to the control extract (39.62 ± 3.76%). Similarly, inhibition of acetylcholinesterase activity increased from 20.91 ± 3.90% (control) to 92.20 ± 3.72% when the tested extract was obtained from T. pinophilus under a combination of suberohydroxamic acid and procainamide. Concluding, increases in antimicrobial activity and acetylcholinesterase inhibition were observed when fungal extracts in the presence of DNA methyltransferases and/or histone deacetylase modulators were tested.

Pectin lyase overproduction by Penicillium griseoroseum mutants resistant to catabolite repression

Abstract Expression of pectinolytic genes is regulated by catabolic repression limiting the production of pectin lyase (PL) if the natural inducer, pectin, is missing from the growth medium. Here, we report the isolation of Penicillium griseoroseum mutants resistant to 2-deoxy-d-glucose (DG) that show resistance to catabolite repression and overproduce PL. Three spontaneous and nine UV-induced mutants were obtained. Some mutants produced sectors (segments morphologically different) that were also studied. The mutants were analyzed for pectinases production on pectinase-agar plates and five mutants and two sectors showing larger clearing zones than the wild type were selected for quantitative assay. Although PL production higher than the wild type has been found, phenotype instability was observed for most of the mutants and, after transfers to nonselective medium, the DG resistance was no longer present. Only mutants M03 and M04 were stable maintaining the DG-resistance phenotype. When growing for 120 h in liquid medium containing glucose with or without pectin, both mutants showed higher PL production. In the presence of glucose as sole carbon source, the mutant M03 produced 7.8-fold more PL than the wild type. Due its phenotypic stability and PL overproduction, the mutant M03 presents potential for industrial applications.

Biodegradation of kerosene: Study of growth optimization and metabolic fate of P. janthinellum SDX7

Penicillum janthinellum SDX7 was isolated from aged petroleum hydrocarbon-affected soil at the site of Anand, Gujarat, India, and was tested for different pH, temperature, agitation and concentrations for optimal growth of the isolate that was capable of degrading upto 95%, 63% and 58% of 1%, 3% and 5% kerosene, respectively, after a period of 16 days, at optimal growth conditions of pH 6.0, 30 °C and 180 rpm agitation. The GC/MS chromatograms revealed that then-alkane fractions are easily degraded; however, the rate might be lower for branched alkanes, n-alkylaromatics, cyclic alkanes and polynuclear aromatics. The test doses caused a concentration-dependent depletion of carbohydrates of P. janthinellum SDX7 by 3% to 80%, proteins by 4% to 81% and amino acids by 8% to 95% upto 16 days of treatment. The optimal concentration of 3% kerosene resulted in the least reduction of the metabolites of P. janthinellum such as carbohydrates, proteins and amino acids with optimal growth compared to 5% and 1% (v/v) kerosene doses on the 12th and 16th day of exposure. Phenols were found to be mounted by 43% to 66% at lower and higher concentrations during the experimental period. Fungal isolate P. janthinellum SDX7 was also tested for growth on various xenobiotic compounds.

Sclerotiorin is non-mutagenic and inhibits human PMNL 5-lipoxygenase and platelet aggregation.

Sclerotiorin, isolated from the fermented broth of Penicillium frequentans, exhibited potent inhibition against human polymorphonuclear leukocytes 5-lipoxygenase and human platelet aggregation with a half maximal value 36 µM and 250 µM, respectively. Further, the Ames test has demonstrated the sclerotiorin to be non-mutagenic.

Microbial mediated preparation, characterization and optimization of gold nanoparticles

The need for eco-friendly and cost effective methods for nanoparticles synthesis is developing interest in biological approaches which are free from the use of toxic chemicals as byproducts. This study aimed to biosynthesize and optimize the size of gold nanoparticles which produced by biotechnological method using Penicillium crustosum isolated from soil. Initially, Penicillium crustosum was grown in fluid czapek dox broth on shaker at 28 ºC and 200 rpm for ten days and then the supernatant was separated from the mycelia to convert AuCl4 solution into gold nanoparticles. The synthesized nanoparticles in the optimum conditions were formed with fairly well-defined dimensions and good monodispersity. The characterizations were done by using different methods (UV-Visible Spectroscopy, Fluorescence, FT-IR, AFM (Atomic Force Microscopy) and DLS (Dynamic Light Scattering). The bioconversion was optimized by Box-Behnken experimental design. The results show that the effective factors in this process were concentration of AuCl4, pH of medium and temperature of shaker incubator. The R² value was calculated to be 0.9999 indicating the accuracy and ability of the polynomial model. It can be concluded that the use of multivariate analysis facilitated to find out the optimum conditions for the biosynthesis of gold nanoparticles induced by Penicillium crustosum in a time and cost effective process. The current approach suggested that rapid synthesis of gold nanoparticles would be suitable for developing a biological process for mass scale production of formulations.

Improvement of submerged culture conditions to produce colorants by Penicillium purpurogenum

Safety issues related to the employment of synthetic colorants in different industrial segments have increased the interest in the production of colorants from natural sources, such as microorganisms. Improved cultivation technologies have allowed the use of microorganisms as an alternative source of natural colorants. The objective of this work was to evaluate the influence of some factors on natural colorants production by a recently isolated from Amazon Forest, Penicillium purpurogenum DPUA 1275 employing statistical tools. To this purpose the following variables: orbital stirring speed, pH, temperature, sucrose and yeast extract concentrations and incubation time were studied through two fractional factorial, one full factorial and a central composite factorial designs. The regression analysis pointed out that sucrose and yeast extract concentrations were the variables that influenced more in colorants production. Under the best conditions (yeast extract concentration around 10 g/L and sucrose concentration of 50 g/L) an increase of 10, 33 and 23% respectively to yellow, orange and red colorants absorbance was achieved. These results show that P. purpurogenum is an alternative colorants producer and the production of these biocompounds can be improved employing statistical tool.

Penicillium species present in Uruguayan salami / Especies de Penicillium presentes en salamines uruguayos

The surface coverage of certain dry fermented sausages such as Italian salami by some species of Penicillium provides their characteristic flavor and other beneficial properties. One of them is the protective effect by means of a uniform film of white mold against undesirable microorganisms. The aim of this work was to identify and to isolate the fungal species present in mature Italian type of salami and to evaluate if it is possible to obtain some of them as starters. In addition, the effects of temperature (14 °C and 25 °C), water activity (a w) (0.90, 0.95 and 0.995) and 2.5 % sodium chloride (NaCl) on fungal growth were determined. Similarly, the proteolytic and lipolytic activity and the ability to produce toxic secondary metabolites were evaluated in order to characterize some possible starter strain. All species found belong to the genus Penicillium, including a performing starter as Penicillium nalgiovense and some potentially toxicogenic species. All the strains showed a higher growth rate at 25 °C. The production of extracellular proteases and lipases was significantly higher at 25 °C than at 14 °C with and without sodium chloride. Only Penicillium expansum produced patulin. On the other hand, Penicillium griseofulvum was the only species that produced ciclopiazonic acid but none of the strains produced penicillin. The species present on salami, Penicillium nalgiovense, Penicillium minioluteum, Penicillium brevicompactum and Penicillium puberulum were unable to produce any of the evaluated toxins. These findings suggest that some fungal isolates from the surface of salami such as P. nalgiovense are potentially useful as starters in sausage manufacture.

La cobertura de la superficie de los embutidos fermentados secos -como el salamín tipo italiano- por algunas especies de Penicillium les proporciona un sabor característico y otras propiedades beneficiosas. Una de ellas es el efecto de protección contra microorganismos indeseables, al formarse una película blanca uniforme de mohos. El objetivo de este trabajo fue aislar e identificar los hongos filamentosos encontrados en la superficie de salamines tipo italiano y evaluar la posibilidad de obtener especies para utilizarse como cultivos iniciadores. Se determinó el efecto de la temperatura, la actividad de agua y del cloruro de sodio sobre el crecimiento fúngico. La actividad proteolítica y lipolítica y la capacidad de producir metabolitos secundarios tóxicos fueron evaluadas con el fin de caracterizar algunos posibles cultivos iniciadores. Todas las cepas fúngicas aisladas e identificadas correspondieron a especies del género Penicillium, algunas benéficas, como Penicillium nalgiovense, y otras potencialmente toxicogénicas. Estas cepas tuvieron diferentes tasas de crecimiento en respuesta a las diferentes condiciones de cultivo. Todas las cepas mostraron mayor crecimiento a 25 °C. La producción de proteasas y lipasas extracelulares fue significativamente mayor a 25 °C que a 14 °C. Penicillium expansum fue la única especie que produjo patulina y Penicillium griseofulvum fue la única que produjo ácido ciclopiazónico. Ninguna de las especies produjo penicilina. Penicillium nalgiovense, Penicillium minioluteum, Penicillium brevicompactum y Penicillium puberulum no produjeron ninguna de las toxinas evaluadas. Estos resultados sugieren que algunos aislamientos fúngicos, como P. nalgiovense, son potencialmente útiles como cultivos iniciadores en la fabricación de estos productos.

Novel antimicrobial secondary metabolites from a Penicillium sp. isolated from Brazilian cerrado soil / Nuevos metabolitos antimicrobianos secundarios de Penicillium sp. aislada en suelo del cerrado de Brasil

The morphological features of a Penicillium, isolated from Brazilian cerrado soil, were characterized and showed to be distinctly different from all well-defined Penicillium species. Chemical and biological investigation on the ethyl acetate extract of this Penicillium isolate resulted in the isolation of three new naphthalenoids: a major metabolite, methyl 6-acetyl-4-methoxy-5,7,8-trihydroxynaphthalene-2-carboxylate and two minor ones, methyl 6-acetyl-4-methoxy-7,8-dihydroxynaphthalene-2-carboxylate and methyl 6-acetyl-4-methoxy-5,8-dihydroxynaphthalene-2-carboxylate. Their structures were determined based on their mono and bidimensional nuclear magnetic resonance data. Acetyl, allyl and methoxyl derivatives of the major metabolite were prepared in order to establish structure-activity relation. Antimicrobial activity of the major natural product and its semi-synthetic derivatives was screened by macro dilution methodology and the corresponding minimum inhibitory concentrations were determined. Natural secondary metabolite methyl 6-acetyl-4-methoxy-5,7,8-trihydroxynaphthalene-2-carboxylate, isolated in a very high yield (0.3175 mg.L-1) showed to be the most active compound, possessing expressive activity against Candida albicans (minimum inhibitory concentration (MIC) 32 ug/mL), Listeria monocitogenes and Bacillus cereus (MIC 64 µg/mL for both).

Decolorization of anthraquinone dye with immobilized Penicillium jensenii.

Cells of Penicillium jensenii were immobilized by entrapment in natural and synthetic polymeric matrices. The decolorization of Reactive Brilliant Blue KN-R by immobilized cells has been studied. It was found that CA-immobilized cells could effectively decolorize reactive brilliant blue KN-R. Many factors affecting the decolorization process were studied, including: pH, temperature, dye concentration, shaker speed and culture time. The reusability of the immobilized cells was evaluated with repeated-batch decolorization experiments. The optimum pH, temperature, shaker speed and culture time of decolorization with CA-, CGN-, and PAA- immobilized cells are 4.0 and 30 degrees C and 150r/min and 48hr respectively, dye concentration could have some effects on decolorization. After four repeated experiments, the decolorization rate of CA-, CGN-, and PAA- immobilized cells could still remain 73.6%, 60.8%, 50.5%, respectively.

Bioleaching of copper from chalcopyrite ore by fungi.

Microorganisms have been geologically active in mineral formation, mineral diagenesis and sedimentation via direct action of their enzymes or indirectly through chemical action of their metabolic products. This property of microorganisms is being harnessed during the recent years for extraction of metals from their ores, especially from low-grade ores. In the present study bioleaching of copper from its low-grade chalcopyrite ore using 26 isolates of acidophilic fungi is reported. Most of these fungal strains belonged to the genera Aspergillus, Penicillium and Rhizopus. The leaching experiments were conducted in Czepek Dox minimal medium containing 1% (100 mesh) ore with shaking at room temperature for 20 days. Out of these, 4 isolates exhibited significant bioleaching activities. Maximum leaching of copper (78 mg/L) was observed with Aspergillus flavus (DSF-8) and Aspergillus niger (DOF-1). Nutritional and environmental conditions for optimum bioleaching were standardized. Present study indicates the usefulness of acidophilic fungi in bioleaching of copper from its low-grade ores.

Sugar-cane juice induces pectin lyase and polygalacturonase in "Penicillium griseoroseum"

The use of other inducers as substitutes for pectin was studied aiming to reduce the production costs of pectin enzymes. The effects of sugar-cane juice on the production of pectin lyase (PL) and polygalacturonase (PG) by "Penicillium griseoroseum" were investigatedd. The Fungus was cultured in a mineral medium (ph 6.3) in a rotary shaker (150 rpm) for 48 h at 25§C. Culture media were supplemented with yeast extract and sucrose or sugar-cane juice. Sugar-cane juice added singly to the medium promoted higher PL activity and mycelical dry weight when compared to pectin and the use of sugar-cane juice and yeast extract or pectin. The results indicated that were similar to those obtained with sucrose-yeast extractor pectin. The results indicated that, even at low concentrations, sugar-cane juice was capable of inducing pectin lyase and polygalacturonase with no cellulase activity in "P. griseoroseum".

Biotransformation of lapachol by Penicillium citreonigrum dierckx: characterization of Lomatiol

Testou-se a biotransformaçäo do lapachol [2-hidroxi-3-(3-metil-2-butenil)-1,4-naftoquinona] por 63 cepas de microorganismos. Com um fungo filamentoso, isolado do solo e identificado como Penicillium citreonigrum Dierckx, observou-se a formaçäo de um metabólito majoritário. Este fungo foi avaliado mais intensamente como biocatalisador para a obtençäo de quantidades suficientes de produtos tranformados para elucidaçäo estrutural. Um produto foi isolado e caracterizado como lomatiol [2-hidroxi-3-(4-hidroxi-3-metil-2-butenil)-1,4-naftoquinona], por análises espectrofotométricas(UV, IV, RMN1H e massa)

Caracterizacao da dose letal minima por irradiacao gama para Pnicillium citrinum / Characterization of minimum lethal dosis of gama irradiation to Penicillium citrinum

O uso das radiacoes ionizantes na destruicao de microrganismos responsaveis pela deterioracao de alimentos ou causadores de infeccoes ou toxinfeccoes alimentares, constituiu-se aplicacao da energia nuclear, para fins verdadeiramente pacificos. Penicillium citrinum e um fungo produtor de micotoxinas, responsaveis por intoxicacoes em humanos e animais que se utilizam de alimentos contaminados. Ha escassez de informacoes sobre a resistencia de P. citrinum a irradiacao gama; assim esta pesquisa objetivou determinar a dose letal por irradiacao gama para esse microrganismo....

Characterization of an aggregated polymer from Penicillium sp. (PB-73 strain)

1. We describe a polymeric of magnesium ammonium phosphate associated with linoleic acid and a basic protein produced by Penicillium sp (strain PB-73). 2. Chemical and physical data suggest a structure characterized as an aggregated polymeric form of protein magnesium ammonium phospholinoleate anhydride. 3. The aggregated polymer has an approximate molecular weight of 316 kDa and a phosphate/magnesium ratio of 2.34

Effect of various organic compounds on synthesis of glucoamylase by an isolated strain Penicillium italicum.

Yeast extract (0.5%) stimulates the production of glucoamylase and cell synthesis while methylene blue (0.1 mM) activates the synthesis of glucoamylase. Studies on the metabolic changes during fermentation of glucoamylase in a selected medium by P. italicum show that the rate of production of glucoamylase and cellular growth are greatly accelerated between 48 and 168 hr. Rapid growth of cells during this period may account for enhanced utilization of maltose and NH4NO3 from the medium. The acid production remains constant from 48 to 144 hr. Different forms of nitrogen decrease steadily. Although methylene blue stimulates the production of glucoamylase in the broth it has practically no effect on the rate of utilization of amino and total nitrogen from the broth.