Bioactive Metabolites Produced by Penicillium sp.1 and sp.2, Two Endophytes Associated with Alibertia macrophylla (Rubiaceae)

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

Penicillium expansum versus antagonist yeasts and patulin degradation in vitro

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeito de produtos alternativos para o controle do bolor verde (Penicillium digitatum) em pós-colheita de citros

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Production of transgalactosylated oligosaccharides (TOS) by galactosyltransferase activity from Penicillium simplicissimum

Ingestion of transgalactosylated oligosaccharides (TOS) and other non-digestible oligosaccharides (NDOs) induces a significant increase in Bifidobacterium, Lactobacillus and some desirable species of Streptococcus populations in the gut of human and other animals (prebiotic effect). This change in the intestinal flora is responsible for several beneficial physiological effects such as a decrease of putrefactive products in the feces, lower blood cholesterol content, higher Ca2+ absorption, a smaller loss of bone tissue in ovariotomized female rats and a lower incidence of colon cancer. beta-Galactosidase from Penicillium simplicissimum, a strain isolated from soil, showed high galactosyltransferase activity when incubated with a highly concentrated lactose solution. Optimum pH and temperature ranges for hydrolytic activity were 4.0-4.6 and 55-60 degrees C, respectively, for a lactose concentration of 5.0% (w/v). Maximal galactosyltransferase activity was obtained at pH 6.5 and 50 degrees C and TOS synthesis was positively associated with lactose concentration in the reaction medium. Thus, when 50 ml of a 60% (w/v) lactose solution was incubated with 26.6 U of beta-galactosidase under the best pH and temperature conditions for transferase activity, a final product with 30.5% TOS (183 mg ml(-1)), 27.5% residual lactose and 42.0% monosaccharides was obtained. (C) 1999 Elsevier B.V. Ltd. All rights reserved.

Purification of an exopolygalacturonase from Penicillium viridicatum RFC3 produced in submerged fermentation

An exo-PG obtained from Penicillium viridicatum in submerged fermentation was purified to homogeneity. The apparent molecular weight of the enzyme was 92 kDa, optimum pH and temperature for activity were pH 5 and 50-55°C. The exo-PG showed a profile of an exo-polygalacturonase, releasing galacturonic acid by hydrolysis of pectin with a high degree of esterification (D.E.). Ions Ca 2+ enhanced the stability of enzyme and its activity by 30%. The K m was 1.30 in absence of Ca 2+ and 1.16mg mL -1 in presence of this ion. In relation to the Vmax the presence of this ion increased from 1.76 to 2.07 μmol min -1mg -1. Copyright © 2009 Eleni Gomes et al.

Production of pectate lyase by penicillium viridicatum RFC3 in solid-state and submerged fermentation

Pectate lyase (PL) was produced by the filamentous fungus Penicillium viridicatum RFC3 in solid-state cultures of a mixture of orange bagasse and wheat bran (1 : 1 w/w), or orange bagasse, wheat bran and sugarcane bagasse (1 : 1 : 0.5 w/w), and in a submerged liquid culture with orange bagasse and wheat bran (3%) as the carbon source. PL production was highest (1,500U mL -1 or 300Ug -1 of substrate) in solid-state fermentation (SSF) on wheat bran and orange bagasse at 96 hours. PL production in submerged fermentation (SmF) was influenced by the initial pH of the medium. With the initial pH adjusted to 4.5, 5.0, and 5.5, the peak activity was observed after 72, 48, and 24 hours of fermentation, respectively, when the pH of the medium reached the value 5.0. PL from SSF and SmF were loaded on Sephadex-G75 columns and six activity peaks were obtained from crude enzyme fromSSF and designated PL I, II, III, IV, V, andVI, while five peaks were obtained fromcrude enzyme fromSmF and labeled PL I', II', III', IV', and VII'. Crude enzyme and fraction III from each fermentative process were tested further. The optimum pH for crude PL from either process was 5.5, while that for PL III was 8.0. The maximum activity of enzymes from SSF was observed at 35°C, but crude enzyme was more thermotolerant than PL III, maintaining its maximum activity up to 45°C. Crude enzyme from SmF and PL III' showed thermophilic profiles of activity, with maximum activity at 60 and 55°C, respectively. In the absence of substrate, the crude enzyme from SSF was stable over the pH range 3.0-10.0 and PL III was most stable in the pH range 4.0-7.0. Crude enzyme from SmF retained 70%-80% of its maximum activity in the acid-neutral pH range (4.0-7.0), but PIII showed high stability at alkaline pH (7.5-9.5). PL from SSF was more thermolabile than that from SmF. The latter maintained 60% of its initial activity after 1 h at 55°C. The differing behavior of the enzymes with respect to pH and temperature suggests that they are different isozymes. Copyright © 2010 Viviani Ferreira et al.

Avaliação do potencial antagônico de leveduras, visando biocontrole de deterioração por penicillium expansum

Considerable losses during apple fruit storage occur due to microbiological diseases, mainly caused by Penicillium expansum, which in addition to fruit pulp deterioration produces patulin, a mycotoxin with carcinogenic and teratogenic activity. Biological control of post-harvest disease by antagonist yeasts focused on killer toxins is an appreciable alternative to the chemical fungicides, due to the low possibility of toxic residues demonstrated during fermentative processes. Twenty out of 44 yeasts (16 isolated from fruits, 10 from corn silage and 18 from laboratory anthill), showed antagonism against spores of P. expansum. The assay in solid medium pointed the strongest nutrient competition antagonism by D. hansenii strain C1 (31 mm inhibition diameter), while D. hansenii strain C7 (15 mm) showed higher antibiosis and parasitism pattern. In the following step the extracellular activity was tested performing the assay with culture supernatant in Yeast Medium agar, where C. guilliermondii P3 was more effective against conidia germination (inhibition rate of 58.15%) while P. ohmeri showed better inhibition on micelial growth (66.17%). The antibiosis showed by both yeasts could suggest probable mechanism associated with killer phenomenon, once both strains were killer positive against sensitive reference strains (S. cerevisiae NCYC 1006 and P. kluyveri CAY-15). In order to enhance the production of antifungal substance, these yeasts were cultivated with P. expansum, but the difference between culture supernatant obtained from yeasts cultivated alone and with mould was not significant (P > 0.05). The results demonstrated that the yeasts application constitute a promising tool, enhancing the biological control of P. expansum in post-harvest diseases of apple fruit.

Purification, characterization, and specificity determination of a new serine protease secreted by penicillium waksmanii

The purpose of this work was to purify a protease from Penicillium waksmanii and to determine its biochemical characteristics and specificity. The extracellular protease isolated that was produced by P. waksmanii is a serine protease that is essential for the reproduction and growth of the fungus. The protease isolated showed 32 kDa, and has optimal activity at pH 8.0 and 35 C towards the substrate Abz-KLRSSKQ-EDDnp. The protease is active in the presence of CaCl2, KCl, and BaCl, and partially inhibited by CuCl2, CoCl2 and totally inhibited by AlCl3 and LiCl. In the presence of 1 M urea, the protease remains 50 % active. The activity of the protease increases 60 % when it is exposed to 0.4 % nonionic surfactant-Triton X-100 and loses 10 % activity in the presence of 0.4 % Tween-80. Using fluorescence resonance energy transfer analysis, the protease showed the most specificity for the peptide Abz-KIRSSKQ-EDDnp with k cat/K m of 10,666 mM-1 s-1, followed by the peptide Abz-GLRSSKQ-EDDnp with a k cat/K m of 7,500 mM -1 s-1. Basic and acidic side chain-containing amino acids performed best at subsite S1. Subsites S2, S3, S′ 2, and S′ 1, S ′ 3 showed a preference for binding for amino acids with hydrophobic and basic amino acid side chain, respectively. High values of k cat/K m were observed for the subsites S2, S3, and S′ 2. The sequence of the N-terminus (ANVVQSNVPSWGLARLSSKKTGTTDYTYD) showed high similarity to the fungi Penicillium citrinum and Penicillium chrysogenum, with 89 % of identity at the amino acid level. © 2012 Springer Science+Business Media New York.

Xylanase and β-Xylosidase from Penicillium janczewskii: Production, Physico-Chemical properties, and application of the crude extract to pulp biobleaching

Extracellular xylanase and β-xylosidase production by a Penicillium janczewskii strain were investigated in liquid cultures with xylan from oat spelts under different physical and chemical conditions. The selected conditions for optimized production of xylanase and β-xylosidase were 7 days, pH 6.5, at 30 °C and 8 days, pH 5.0, at 25 °C, respectively. The xylanase exhibited optimal activity in pH 5.0 at 50 °C and the β- xylosidase in pH 4.0 at 75 °C. The xylanase was more stable at pH 6.0 to 9.5, while the β-xylosidase remained stable at pH ranging from 1.6 to 5.5. The xylanase half-life (T50) at 40, 50, and 60 °C was 183, 15, and 3 min, respectively. β-xylosidase half-life was 144, 8, and 4 min at 50, 65, and 75 °C, respectively. When applied to the biobleaching of Eucalyptus kraft pulp, xylanase dosages of 2 and 4 U/g dried pulp reduced, respectively, kappa number by 3.0 and 3.3 units after 1 h treatment, demonstrating that the use of P. janczewskii xylanases in this process is quite promising. The pulp viscosity was not altered, confirming the absence of cellulolytic enzymes in the fungal extract.

Controle em pós-colheita de Penicillium digitatum em laranja-pera com microrganismos e tratamento térmico

The consumer market demands food without pesticide residues. Therefore, this study focused on evaluating the control of green mold in Pera orange trees with biocontrol agents (Bacillus subtilis, Bacillus licheniformis and Bacillus subtilis (QST 713)), associated or not with heat treatment. The fruit was obtained in packinghouse before processing, being washed and disinfected with the use of Sodium Hypochlorite. Fruits submitted to these treatments were stored from 11 to 28 days at temperature of 10 °C and RH 90%±5 or for eight days at 20 °C and 90%±5. In general, the heat treatment reduced the disease severity determine by the area under the disease progress curve in the fruit and the incidence of natural postharvest disease in Pera oranges. On the other hand, biocontrol agents did not control the disease, showing that the organisms tested did not present curative activity against the green mold.

Avaliação de microrganismos antagônicos, Saccharomyces cerevisiae e Bacillus subtilis para o controle de Penicillium Digitatum

Citrus fruits are affected by diverse diseases, mainly the fungal infections, which affect productivity and quality, especially when it targets the market of fresh fruit. Among the fungal diseases that occur in postharvest, there is the green mold caused by Penicillium digitatum. The control measures are based mainly in the treatment of fruits with different combinations of fungicides in packing-house. Due to restrictions on the presence of residues of fungicides in citrus fruits and the increasing development of resistant strains of pathogens to the fungicide used, it is necessary to search for control alternatives such as biological control. Therefore, this study aimed to: (i) verify the antagonistic effect of biological control agents (BCA), being 13 isolates of Bacillus subtilis and 06 isolates of Saccharomyces cerevisiae against P. digitatum, (ii) study in vitro interactions between pathogen and BCA (iii) determine the effect of integration of antagonists with sodium bicarbonate and carnauba wax in the control of green mold. The results showed that the majority of the isolates, and all yeast isolates inhibited the mycelial growth of the phytopathogen. Only one isolate of B. subtilis (ACB-84) was able to inhibit the germination of P. digitatum (72% of inhibition), whereas ACB-K1 and ACB-CR1 (S. cerevisiae) were the most effective with inhibition from 78 and 85.7% respectively. The addition of sucrose (0.5%) favored the inhibition of conidia germination by the yeast isolates. The results from the in vivo control showed the viability of S. cerevisiae ACB-K1 and ACB-CR1 to control P. digitatum in 'Tahiti' lime fruits and orange 'Hamlin' fruits, respectively. The combination of sodium bicarbonate and biocontrol agents did not result in improvements in the curative control of the green mold. Carnauba wax (18% of TSS) favored the antagonistic activity of S. cerevisiae, and this effect depended on the variety of fruits in the study and of the yeast isolate used for the biocontrol.

Production, purification, and characterization of a major penicillium glabrum xylanase using brewer's spent grain as substrate

In recent decades, xylanases have been used in many processing industries. This study describes the xylanase production by Penicillium glabrum using brewer's spent grain as substrate. Additionally, this is the first work that reports the purification and characterization of a xylanase using this agroindustrial waste. Optimal production was obtained when P. glabrum was grown in liquid medium in pH 5.5, at 25 °C, under stationary condition for six days. The xylanase from P. glabrum was purified to homogeneity by a rapid and inexpensive procedure, using ammonium sulfate fractionation and molecular exclusion chromatography. SDS-PAGE analysis revealed one band with estimated molecular mass of 18.36 kDa. The optimum activity was observed at 60 °C, in pH 3.0. The enzyme was very stable at 50 °C, and high pH stability was verified from pH 2.5 to 5.0. The ion Mn2+ and the reducing agents β-mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg2+, Zn2+, and Cu2+ as well as the detergent SDS were strong inhibitors of the enzyme. The use of brewer's spent grain as substrate for xylanase production cannot only add value and decrease the amount of this waste but also reduce the xylanase production cost. © 2013 Adriana Knob et al.

Produção de enzimas celulolíticas pelos fungos thermoascus aurantiacus CBMAI 756, thermomyces lanuginosus, Trichoderma reesei QM9414 e Penicillium viridicatum RFC3 e aplicação na sacarificação do bagaço de cana de açucar com diferentes pré-tratamentos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Potencial anti-inflamatório de Pyrenocine A isolada do Penicillium paxilli: abordagem profilática e terapêutica

Pós-graduação em Biociências e Biotecnologia Aplicadas à Farmácia - FCFAR

Produção de biossurfactantes pelos fungos Aspergillus ochraceus e Penicillium expansum em fermentação semi-sólida utilizando resíduos agroindustriais como substrato

Pós-graduação em Microbiologia - IBILCE