Potent Anti-Inflammatory Activity of Pyrenocine A Isolated from the Marine-Derived Fungus Penicillium paxilli Ma(G) K

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Produção e caracterização de a-L-arabinofuranosidase de Penicillium janczewskii

A xilana, o segundo principal componente da parede celular vegetal, pertence ao grupo das hemiceluloses, sendo composta por diferentes carboidratos, principalmente xilose e arabinose. Na natureza, devido a sua heterogeneidade estrutural, este complexo polissacarídeo é completamente hidrolisado pela ação sinergística de diferentes enzimas, incluindo xilanases e β-xilosidases, responsáveis pela degradação da sua cadeia principal e outras enzimas chamadas auxiliares ou desramificantes, importantes para remoção dos grupos laterais. Dentre estas últimas, destacam-se as α-L-arabinofuranosidases, enzimas responsáveis pela remoção de resíduos L-arabinofuranosil do polímero. As arabinofuranosidases podem ser produzidas por micro-organismos, como bactérias e fungos, sendo essencialmente enzimas extracelulares. Fungos, especialmente os de solo e madeira, têm sido utilizados para produção de enzimas xilanolíticas, sendo particularmente interessantes do ponto de vista industrial, pelo fato de secretarem suas enzimas diretamente no meio em que se encontram não necessitando de ruptura celular para a liberação das mesmas. Além disso, apresentam níveis de produção mais elevados que os obtidos em culturas bacterianas ou de leveduras. A crescente preocupação com a escassez dos recursos naturais e com a degradação ambiental tem levado à busca por tecnologias mais eficientes, mais competitivas e menos poluentes. Atualmente, muitos processos industriais empregam enzimas microbianas, apresentando inúmeras vantagens em relação às técnicas convencionais. As α-L-arabinofuranosidases podem ser utilizadas individualmente ou em combinação com outras enzimas, representando uma ferramenta promissora para aplicação em diversos processos biotecnológicos como no branqueamento da polpa celulósica, na síntese de oligossacarídeos, na produção de etanol de segunda geração, ou ainda...(Resumo completo, clicar acesso eletrônico abaixo)

Influência de parâmetros nutricionais e físicos sobre a produção de Lipase por Penicillium janthinellum

As lipases (triacilglicerol acil hidrolases, E.C. 3.1.1.3) constituem uma classe de enzimas que catalisam a hidrólise de ligações ésteres dos triacilgliceróis de cadeia longa, operando na interface óleo/água. Em meios orgânicos, dependendo da quantidade de água, essas enzimas podem catalisar reações de esterificação e transesterificação, atuando como biocatalisadores da síntese de óleos e gorduras. As lipases destacam-se em processos industriais, nos quais são utilizadas como aditivos na produção de detergentes, de papel e celulose, na indústria de laticínios, de cosméticos, entre outros. As lipases microbianas são de maior interesse para aplicação em biotecnologia e em química orgânica, sendo necessária a prospecção de novas fontes de lipases com propriedades distintas. O presente trabalho avaliou a influência de diferentes meios de cultura, do tempo de cultivo, de fontes de carbono e sua concentração, de fontes de nitrogênio, agitação, pH e temperatura sobre a produção de lipase extracelular pelo fungo filamentoso Penicillium janthinellum, isolado de solo de região de Mata Atlântica. A avaliação das melhores condições de cultivo foi realizada através da análise da atividade enzimática, acompanhada pela determinação do íon ρ-nitrofenol liberado na hidrólise do substrato sintético ρ- nitrofenil palmitato. Os experimentos foram realizados em frascos de Erlenmeyer de 125mL contendo 25mL de meio de cultivo, inoculados com 1mL de suspensão contendo 107 conídios, e incubados a 28°C com agitação de 160 rpm e pH 5,5. Entre os quatro meios de cultura testados, o melhor resultado (0,476 ± 0,04 U/mL), com 5 dias de cultivo, foi verificado com o meio 2 composto por (g/L): bacto-peptona 5,0, extrato de levedura 1,0, NaNO3 0,5, KCl 0,5, MgSO4·7H2O 0,5, KH2PO4 2,0 e azeite de oliva 10,0)... (Resumo completo, clicar acesso eletrônico abaixo)

Improvement of submerged culture conditions to produce colorants by Penicillium purpurogenum

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Extraction of natural red colorants from the fermented broth of Penicillium purpurogenum using aqueous two-phase polymer systems

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Purificação, imobilização e caracterização bioquímica de lipase produzida por Penicillium sect. Gracilenta CBMAI 1583 em cultivo submerso

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Partitioning and extraction of collagenase from Penicillium aurantiogriseum in poly(ethylene glycol)/phosphate aqueous two-phase system

Purification of collagenase produced by Penicillium aurantiogriseum URM4622 was carried using a PEG/phosphate aqueous two-phase system (ATPS). A 2(3)-full experimental design was used to investigate the influence of PEG molar mass, PEG concentration and phosphate concentration on the selected responses, namely partition coefficient, activity yield and purification factor. The ATPS was composed of PEG (molar mass of 550, 1500 and 4000 g/mol) at concentrations of 15.0, 17.5 and 20.0% (w/w) and phosphate at concentrations of 12.5, 15.0 and 17.5% (w/w). The best results of one-step extraction of collagenase from the fermentation broth (partition coefficient of 1.01, activity yield of 242% and purification factor of 23.5) were obtained at pH 6.0 using 20.0% (w/w) PEG 550 and 17.5% (w/w) phosphate. The results of this preliminary study demonstrate that the selected ATPS is satisfactorily selective for the extraction of such a collagenase. (C) 2012 Elsevier B.V. All rights reserved.

Preliminary Characterization of Novel Extra-cellular Lipase from Penicillium crustosum Under Solid-State Fermentation and its Potential Application for Triglycerides Hydrolysis

Current studies about lipase production involve the use of agro-industrial residues and newly isolated microorganisms aimed at increasing economic attractiveness of the process. Based on these aspects, the main objective of this work is to perform the partial characterization of enzymatic extracts produced by a newly isolated Penicillium crustosum in solid-state fermentation. Lipase extract presented optimal temperature and pH of 37 A degrees C and 9-10, respectively. The concentrated enzymatic extract showed more stability at 25 A degrees C and pH 7. The enzymes kept 100% of their enzymatic activity until 60 days of storage at 4 and -10 A degrees C. The stability under calcium salts indicated that the hydrolytic activity presented decay with the increase of calcium concentration. The specificity under several substrates indicated good enzyme activities in triglycerides from C4 to C18.

Biosynthesis of Two Dihydropyrrole-Polyketides from a Marine-Derived Penicillium citrinum

Feeding experiments with C-13-labeled precursors were performed in order to establish the biosynthesis of two N-acylated dihydropyrroles, (8E)-1-(2,3-dihydro-1H-pyrrol-1-yl)-2- methyldec-8-ene-1,3-dione (1) and 1-(2,3-dihydro-1H-pyrrol-1-yl)-2- methyldecane-1,3-dione (2), isolated from the cultures of a marine-derived Penicillium citrinum. The biosynthesis of both, 1 and 2, involves the incorporation of acetate, methionine and ornithine.

Metabolic engineering of beta-oxidation in Penicillium chrysogenum for improved semi-synthetic cephalosporin biosynthesis

Industrial production of semi-synthetic cephalosporins by Penicillium chrysogenum requires supplementation of the growth media with the side-chain precursor adipic acid. In glucose-limited chemostat cultures of P. chrysogenum, up to 88% of the consumed adipic acid was not recovered in cephalosporinrelated products, but used as an additional carbon and energy source for growth. This low efficiency of side-chain precursor incorporation provides an economic incentive for studying and engineering the metabolism of adipic acid in P. cluysogenum. Chemostat-based transcriptome analysis in the presence and absence of adipic acid confirmed that adipic acid metabolism in this fungus occurs via beta-oxidation. A set of 52 adipate-responsive genes included six putative genes for acyl-CoA oxidases and dehydrogenases, enzymes responsible for the first step of beta-oxidation. Subcellular localization of the differentially expressed acyl-CoA oxidases and dehydrogenases revealed that the oxidases were exclusively targeted to peroxisomes, while the dehydrogenases were found either in peroxisomes or in mitochondria. Deletion of the genes encoding the peroxisomal acyl-CoA oxidase Pc20g01800 and the mitochondrial acyl-CoA dehydrogenase Pc20g07920 resulted in a 1.6- and 3.7-fold increase in the production of the semi-synthetic cephalosporin intermediate adipoyl-6-APA, respectively. The deletion strains also showed reduced adipate consumption compared to the reference strain, indicating that engineering of the first step of beta-oxidation successfully redirected a larger fraction of adipic acid towards cephalosporin biosynthesis. (C) 2012 Elsevier Inc. All rights reserved.

Biosynthesis of two dihydropyrrole-polyketides from a marine-derived Penicillium citrinum

Feeding experiments with 13C-labeled precursors were performed in order to establish the biosynthesis of two N-acylated dihydropyrroles, (8E)-1-(2,3-dihydro-1H-pyrrol-1-yl)-2-methyldec8-ene-1,3-dione (1) and 1-(2,3-dihydro-1H-pyrrol-1-yl)-2-methyldecane-1,3-dione (2), isolated from the cultures of a marine-derived Penicillium citrinum. The biosynthesis of both, 1 and 2, involves the incorporation of acetate, methionine and ornithine.

Productos naturales de Schizochytrium aggregatum, Paecilomyces variotii y Penicillium roqueforti, hongos de interés acuícola

[ES]El estudio químico de un organismo, y la elucidación de las estructuras moleculares de sus componentes, puede dar lugar a descubrimientos que aconsejen una posterior producción industrial del mismo. Con este propósito, se seleccionaron tres microorganismos para ser investigadas químicamente, siendo estos: un hongo zoospórico, Schizochytrium aggregatum, y dos hifomicetos, Paecilomyces variotii y Penicillium roqueforti, estos últimos fueron aislados del medio marino del litoral canario. Los resultados demuestran que los tres hongos estudiados no son, en nuestras condiciones de cultivo, productores de micotoxinas; al contrario, producen importantes componentes nutricionales, lo cual supone que se pueden utilizar directamente para la nutrición animal en acuicultura. Por otra parte, también se pueden usar para producir industrialmente productos de alto valor añadido como oleína, peróxido de ergosterol, (9,11)-dehidroperóxido del ergosterol, p-hidroxi-benzaldehido, D- (-)-manitol y 2-deoxi-2-fosfamino-?-D-glucopyranosa. Esta última es nueva en la bibliografía

Enzimi, acidi organici ed altri metaboliti coinvolti nella patogenesi di Penicillium spp

Blue mould caused by Penicillium expansum Link is one of the most destructive rot of pome fruit in all growing areas (Snowdon, 1990; Jones and Aldwinckle, 1991; Tonini,1996) In the past, Penicillium rot has been controlled by fungicide postharvest treatment mainly by thiabendazole (TBZ) and benomyl (Hardenburg and Spalding, 1972), but their intense use produced the appearance of resistant strains with a great reduction of their activity The aims of the present study were to characterize the isolates of Pencillium sp causing blue mold on pear in Italy by physiological and biochemical parameters. In particular differencing also the behavior of isolates to relationship with sensitivity or resistance to TBZ treatments. We have examined the early stage of infection in relation to enzyme activity, local modulation of pH, production of organic acids, and to secondary metabolism of pathogen. The results described here confirm that the majority of P. expansum isolates from pears packing houses are resistant to TBZ, Among the TBZ-resistant isolates scored in this work, different isolates (RR) showed higher percentage of conidial germination on TBZ-amended medium compared to non amended medium. This may indicate a stimulatory effect of TBZ on conidial germination. Therefore TBZ treatments are not only ineffective for controlling P. expansum, but they may also increase the severity of blue mould on fruits. In the absence of fungicide, isolates showed a significant difference for infection severity, R and RR isolates are characterized by higher pathogenic fitness on fruits, producing larger lesions than S isolates. These data are supported by the study with laboratory-induced resistant isolates, which shows the lack of correlation between TBZ resistance and osmotic sensitivity, and highlights the association between TBZ resistance and infection severity (Baraldi et al 2003). Enzymatic screening gave a positive reaction to esterase, urease, pectinase activity, in addition, the pathogen is able to synthesize a complex enzyme act to degrade the main components of the cell wall especially pectin and cellulose. Isolated sensitive and resistant are characterized by a good activity of pectinase, especially from poligactoronase, which, as already reported by several studies (D'hallewin et al, 2004; Prusky et al, 2004), are the basis of degradative process of cell wall. Also, although the measure was minor also highlighted some activities of cellulase, but even note in the production of this kind of cellulase and hemicellulase P. Expansum were not targeted, studies have found no other source of information in this regard. Twenty isolates of Penicillium expansum, were tested in vitro ad in vivo for acid production ability and pH drop. We have found that modulation of pH and the organic acids extrusion were influence to various parameter:  Initial pH: in general, the greatest reduction of pH was observed in isolates grown at pH 7, except for four isolates that maintained the pH of the medium close to 7, the others significantly decreased the pH, ranging from 5.5 to 4.1.. In extreme acid condition (pH 3,0) growth and modulation of pH is most lower respect optimal condition (pH 5,0). Also isolates R and RR have showed a greater adaptation to environmental condition more than isolates S.  Time: although the acidification continues for some days, PH modulation is strongest in early hours (48-72 hours)of inoculation process. Time also affects the quality of organic acids, for example in vitro results showed an initial abundant production of succinc acid, followed to important production of galacturoinc acid.  Substrates: there are many differences for the type of acids produced in vitro and in vivo. Results showed in vivo an abundant production of galacturonic, malic, and citric acids and some unknown organic acids in smaller concentrations. Secondary metabolite analysis revealed intra-specific differences, and patulin was found in all isolates, but most significant reduction was observed between in vitro and in vivo samples. There was no correlation between the concentration of patulin, and the percentage of infected fruits, but sample with a lower infection severity of rotten area than the others, showed a significantly lower mycotoxin concentration than samples with a higher lesion diameter of rotten area. Beyond of patulin was detected the presence of another secondary metabolite, penitrem A.

Study of thiabendazole resistance and volatile organic compounds production of Penicillium expansum strains

The years of excessive use of thiabendazole to control Penicillium expansum has induced the development of resistance. Sensitivity of fourty eight strains collected from orchards and packinghouses in Emilia Romagna to pure and commercial TBZ was determined in vitro on TBZ amended medium (400μg/mL). Out of 48 strains, 35 were thiabendazole-sensitive (S) and 13 were thiabendazole-resistant (R). Microtiter assay adapted to P. expansum, showed EC50 values ranging from 54 to 320 μg/mL for ten TBZ-resistant strains. At the highest dose (50 μg/mL), resistant strains growth was not inhibited and the reported MICs value were >1000 μg/mL. Therefore, preliminary screening combined with microtiter assay, can be a good strategy to test susceptibility to TBZ. Mutations in the β-tubulin gene were studied on amino acid sequences from residue 167 to residue 357 of 10 P. expansum strains. Mutation at codon 198 was associated with TBZ-resistance. However, its absence in 3 resistant strains can be explained by the involvement of other mechanisms. Moreover, a P. expansum strain LB8/99 showed good antifungal effect against some fungal pathogens through double petri dish assay. It inhibited both mycelium growth and conidia germination of B. cinerea, C. acutatum, and M. laxa, and reduced significantly by 53% and 18% respectively P. expansum. Three major VOCS: geosmin, phenethyl alcolhol (PEA) and an unknown substance were identified by GC-MS analysis. Consistent fumigation of fungal pathogens with PEA (1230 mg/mL), inhibited both conidia germination and mycelium growth of all pathogens, except conidia germination of P. expansum that was reduced by 90% with respect to control. While, the concentration of PEA produced naturally by LB8/99 was ineffective in controlling the pathogens and seemed to have a synergic or additive effect with the other VOCS. Investigations to study the biofumigant effect of LB8/99 on other commodities like seeds and seedlings are in progress.