Taxonomic assessment and enzymes production by yeasts isolated from marine and terrestrial Antarctic samples

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Understanding the Function of Conserved Variations in the Catalytic Loops of Fungal Glycoside Hydrolase Family 12

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Leaf-cutting ants: an unexpected microenvironment holding human opportunistic black fungi

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

Yeasts found on an ephemeral reproductive caste of the leaf-cutting ant Atta sexdens rubropilosa

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comparative study of toxicity of azo dye Procion Red MX-5B following biosorption and biodegradation treatments with the fungi Aspergillus niger and Aspergillus terreus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Transport of amino acids from milk whey by Caco-2 cell monolayer after hydrolytic action of gastrointestinal enzymes

The bioavailability of amino adds from milk whey protein hydrolysates was evaluated using diffusion of the substances through semi-permeable membranes (dialyzability) and transport by Caco-2 cell cultures. The hydrolysates with low degree of hydrolysis (LDH) and high degree of hydrolysis (HDH) were obtained after 120 min of reaction time at 50 degrees C after the initial addition of pepsin, followed by the addition of trypsin, chymotrypsin and carboxypeptidase-A. The proteins and hydrolysates were further subjected to in vitro digestion with pepsin plus pancreatin. HPLC was used to determine the concentrations of dialyzable amino adds (48.4% of the non-hydrolyzed proteins, 63.2% of the LDH sample and 58.3% of the HDH sample), demonstrating the greater dialyzability of the hydrolysates. The LDH and HDH whey protein hydrolysates prepared with pepsin, trypsin, chymotrypsin and carboxypeptidase-A showed only 14.7% and 20.8% of dialyzable small peptides and amino acids, respectively. The efficiency of absorption was demonstrated by the preferential transport of Ile, Lou and Arg through a layer of cells. In the LDH hydrolysate, Tyr was also transported. Prior high- and low-degree hydrolysis of the whey provided transport by 5.7% and 6.6%, respectively, in comparison with 23% for non-hydrolyzed proteins, considering the total amount of these amino adds that was applied to the cells. (C) 2014 Elsevier Ltd. All rights reserved.

Antifungal Activity of Decyl Gallate against Several Species of Pathogenic Fungi

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

Biodegradação de materiais poliméricos por fungos filamentosos

The use of polymeric materials has grown in recent years due to its high durability features, atoxicity, shaping versatility and environment resistance. However, while these features represent good advantages to several industry sectors, it results in one of the most serious environmental problems of contemporary society: the rising accumulation of these material, mainly due to the inadequate disposal of waste. Meanwhile, in order to minimize this problem, some mitigation techniques comes up (arises), among which the use of biodegradable polymers has been gaining attention. Because of their easily action of microorganisms, such material degrade more rapidly, becoming integrated to nature. Furthermore, due to the fact of biodegradation is a natural process, occurring through the action if micro-organisms in the environment itself, it is considered the “cleaner” alternative found so far to plastic components reincorporation in the nature. Among the micro-organisms capable of biodegradation process are the filamentous fungi. These micro-organisms have many advantages over the others, the major one being the capacity to produce a range of enzymes capable of degrading different materials. In this context, the present review made it possible to see the importance of this process as an agent of environmental preservation, suggest the use of blends to minimize the problems of cost and flexibility of biodegradable polymeric materials, as well as noting the lack of studies related to this subject nowadays

Phenotypic expression of homozygous hemoglobin S in relation to β-globin haplotypes, glutathione S-transferase polymorphisms and detoxification enzymes

Sickle cell anemia (SCA) shows a pathophysiology that involves multiple changes in sickle cell erythrocytes, vaso-occlusive episodes, hemolysis, activation of inflammatory mediators, endothelial cell dysfunction, and oxidative stress. These events complicate treatment and culminate in the development of manifestations such as anemia, pain crises and multiorgan dysfunction. The aim of this study was to evaluate, in SCA patients, oxidative stress and antioxidant capacity markers, correlating them to treatment with hydroxyurea (HU), β-globin haplotypes and glutathione S-transferase polymorphisms (GSTT1, GSTM1 and GSTP1), in comparison to a control group (CG). The study groups were composed of 48 individuals without hemoglobinopathies (CG), SCA patients treated with HU [AF (+HU), N = 13] and untreated SCA patients [AF (-HU), N = 15], after informed consent. The groups were analyzed using cytological, electrophoretic, chromatographic and molecular methods and information from medical records. The GSTM1 and GSTT1 polymorphisms were determined by multiplex PCR, while the GSTP1 polymorphism by PCR-RFLP. Biochemical parameters were measured using spectrophotometric methods [TBARS, TEAC and catalase (CAT) and GST activities] and a chromatographic method [glutathione (GSH)]. The fetal Hb (Hb F) levels observed in the SCA (+HU) group (10.9%) confirmed the already well-described pharmacological effect of HU, but the SCA (-HU) group also had high Hb F levels (6.1%), which may have been influenced by genetic factors not targeted in this study. We found a higher frequency of the Bantu haplotype (48.2%), followed by the Benin (32.1%) and also Cameroon haplotypes, rare in our population, and 19.7% of atypical haplotypes. The presence of Bantu haplotype was related to higher lipid peroxidation levels in patients, but also, it conferred a differential response to HU treatment, raising Hb F levels in 52.6% (P = 0.03). The protective effect of Hb F was confirmed, because the increase in their levels resulted in a 41.3% decrease in lipid peroxidation levels (r = -0.74, P = 0.0156). The genotypic frequency of the GST polymorphisms observed was similar to that of other studies in the Brazilian population, and its association with biochemical markers revealed a significant difference only for the GSTP1 polymorphism, where patients with genotype V/V showed higher GSH and TEAC levels (P = 0.04 and P = 0.03, respectively) compared to patients with genotype I/I. The TBARS levels were about five to eight times higher in the SCA (+HU) and SCA (-HU) groups, respectively, compared to controls, and HU produced a 35.2% decrease in lipid peroxidation levels in the SCA (+HU) group (P < 0.0001). Moreover, the SCA (+HU) group showed higher TEAC levels when compared to CG (P = 0.002). We did not find any significant difference in GST activity between the groups studied (P = 0.76), but CAT activity was about 17 and 30% lower in SCA (+HU) and SCA (-HU) groups, respectively (P < 0.00001). Plasma GSH levels were ~2 times higher in SCA patients than in the control group (P = 0.0005) and showed a positive correlation with TBARS levels, confirming its antioxidant function. HU treatment contributed to higher CAT activity and TEAC levels and lower lipid peroxidation, and its pharmacological effect showed a “haplotype-dependent” response. These findings may contribute to elucidating the potential of HU in ameliorating oxidative stress in SCA subjects.

Fungos derivados da Antártica: biodiversidade e produção de enzimas lignocelulóticas a baixas e médias temperaturas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Identification of quinolactin alkaloids from fungi associated to Brazilian red algae Dichotomaria marginata by LC-PDA-MS and LC-MSn

Fungi isolated from marine organisms have been shown to produce several interesting secondary metabolites with important biological activities. Such chemical diversity may be associated to environmental stress conditions and may represent an important source of NCE for bioprospection. Quinolactins belong to a rare fungi-alkaloid class with a unique N-methyl-quinolone moiety fused to a lactam ring and present several bioactivities1. Fungi strain Dm1 was isolated from red alga Dichotomaria marginata, collected from Brazil SE coast, and was grown in sterile rice solid media at 26oC 2, which was then extracted with MeOH. The MeCN fr. from the MeOH extract was chromatographed over Sephadex LH-20 and fr. 4 afforded quinolactin (QL) alkaloids B1, B2 and A, whereas fr. 5 afforded quinolactin D1 after purification by HPLC-DAD. Structural determination of pure compounds was based on HRMS, UV, and NMR spectral analyses, in addition to comparison with literature data and Antimarin® databank. UV data indicated the presence of similar chromophores with λmax at ca. 247 and 320nm. HRMS and tandem MS analyses using both negative and positive ion modes for the isolated compounds indicated their molecular formula and structural features, as for QL B1: C15H16O2N2 [M+H 257], which showed one fragment at m/z 214 [-CHNO]; QL B2: C15H16O3N2 [M+H 273], with product ions at m/z 230 [-CHNO.] and m/z 186 [-C4H9NO.]; for QL A: C16H18N2O2 [M+H 271], which presented one ion at m/z 214, due to loss of fragment (-C4H9) from the molecular ion; and for QL D1: C16H18N2O3 [M+H 287], with product ions at m/z 186 [-CHNO] and m/z 230 [-C4H9]. Such data suggested fragmentation proposals, e.g. for Quinolactin B1 (Fig. 1), which confirmed the structures of the isolated quinolactins, and may represent an important contribution for the sustainable exploration of marine biodiversity.

Production of active recombinant eIF5A: reconstitution in E. coli of eukaryotic hypusine modification of eIF5A by its coexpression with modifying enzymes

Eukaryotic translation initiation factor 5A (eIF5A) is the only cellular protein that contains the polyamine-modified lysine, hypusine [Nε-(4-amino-2-hydroxybutyl)lysine]. Hypusine occurs only in eukaryotes and certain archaea, but not in eubacteria. It is formed post-translationally by two consecutive enzymatic reactions catalyzed by deoxyhypusine synthase (DHS) and deoxyhypusine hydroxylase (DOHH). Hypusine modification is essential for the activity of eIF5A and for eukaryotic cell proliferation. eIF5A binds to the ribosome and stimulates translation in a hypusine-dependent manner, but its mode of action in translation is not well understood. Since quantities of highly pure hypusine-modified eIF5A is desired for structural studies as well as for determination of its binding sites on the ribosome, we have used a polycistronic vector, pST39, to express eIF5A alone, or to co-express human eIF5A-1 with DHS or with both DHS and DOHH in Escherichia coli cells, to engineer recombinant proteins, unmodified eIF5A, deoxyhypusine- or hypusine-modified eIF5A. We have accomplished production of three different forms of recombinant eIF5A in high quantity and purity. The recombinant hypusine-modified eIF5A was as active in methionyl-puromycin synthesis as the native, eIF5A (hypusine form) purified from mammalian tissue. The recombinant eIF5A proteins will be useful tools in future structure/function and the mechanism studies in translation.

Identificação e caracterização de fungos melanizados com potencial de degradação de tolueno

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Produção e caracterização das Tanases do fungo filamentoso Aspergillus carbonarius

Pós-graduação em Biotecnologia - IQ