Easily handling penicillin G acylase magnetic cross-linked enzymes aggregates: Catalytic and morphological studies

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

The effect of exogenous fibrolytic enzymes and a ferulic acid esterase-producing inoculant on the fibre degradability, chemical composition and conservation characteristics of alfalfa silage

The objective of this study was to determine the effect of applying fibrolytic enzymes at ensiling, either alone or in combination with a ferulic acid esterase-producing bacterial silage inoculant, on the silage conservation characteristics and nutritive value of alfalfa (Medicago sativa L). Second-cut alfalfa (340 g DM/kg fresh crop) was harvested, wilted, chopped and sub-sampled into 24 batches. Samples were randomly allocated in triplicate to one of four enzyme product treatments supplying endoglucanases and xylanases: none (control), EN1, EN2, EN3; applied alone or in combination with a ferulic acid esterase-producing silage inoculant (FAEI). Treatments were arranged in a 4 x 2 factorial design. All enzyme treatments were applied at 2 ml enzyme product/kg herbage DM, and inoculant was applied at 1 x 10(5) cfu/g fresh herbage. Samples were packed into laboratory-scale silos and stored for 7, 27 or 70 days, and analysed for dry matter (DM) losses, aerobic stability, chemical composition and in vitro ruminal degradability. The use of enzymes did not affect (P>0.05) ensilage DM losses or lactic or acetic acid concentrations after 70 days of ensilage, compared to the control silage. Silage produced using EN1 had lesser neutral detergent fibre (aNDF, P=0.046) and acid detergent fibre (ADF; P=0.006) concentrations than control silage. However, no difference (P>0.05) was observed between the control silage and silage produced with EN1 for aNDF or ADF degradability (NDFD, ADFD). Silages produced with FAEI had greater DM losses (P=0.017) and pH (P<0.001) and lesser NDFD (P=0.019), ADFD (P=0.010) and proportion of lactic acid in the total fermentation products (P=0.006) after 70 days of ensilage, compared to uninoculated silages. The use of fibrolytic enzymes did not have a major effect on the ensilage fermentation of alfalfa, either ensiled alone or with an inoculant. No advantage in ruminal DM or fibre degradability was observed for silages produced with fibrolytic enzymes. The use of a ferulic acid esterase-producing inoculant alone did not improve the nutritive value of alfalfa silage, and did not promote any incremental effects when applied in combination with fibrolytic enzyme products. Crown Copyright (C) 2014 Published by Elsevier B.V. All rights reserved.

Electrochemical studies based on local interfacial ph changes of gold nanoparticles immobilized on polystyrene brushes

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

Effect of initial pH in levan production by Zymomonas mobilis immobilized in sodium alginate

Zymomonas mobilis was immobilized using a cell suspension fixed to 8.6 x 10(7) CFU mL(-1) by spectrophotometry. This biomass was suspended in sodium alginate solution (3%) that was dropped with a hypodermic syringe into 0.2 M calcium chloride solution. Was test two initial pH of fermentation medium (4 and 5) and different sucrose concentrations 15, 20, 25, 30 and 35% at 30 degrees C, without stirring for 24, 48, 72 and 96 hours. The levan production to pH 4 was high in sucrose 25% for 24 (16.51 g L-1) and 48 (15.31 g L-1) hours. The best values obtained to pH 5 was in sucrose 35% during 48 (22.39 g L-1) and 96 (23.5 g L-1) hours, respectively. The maximum levan yield was 40.8% and 22.47% in sucrose 15% to pH 4 and 5, respectively. Substrate consumption to pH 4 was bigger in sucrose 15 (56.4%) and 20% (59.4%) and to pH 5 was in 25 (68.85%) and 35% (64.64%). In relation to immobilization efficiency, Zymomonas mobilis showed high adhesion and colonization in support, indicated by cell growth increased from 107 to 10(9) CFU mL(-1) during fermentation time.

Vitamin B6-Dependent Enzymes in the Human Malaria Parasite Plasmodium falciparum: A Druggable Target?

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

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

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Electrochemical Properties of the Oxo-Manganese-Phenanthroline Complex Immobilized on Ion-Exchange Polymeric Film and Its Application as Biomimetic Sensor for Sulfite Ions

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

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.

Speciation analysis of inorganic arsenic in river water by Amberlite IRA 910 resin immobilized in a polyacrylamide gel as a selective binding agent for As(v) in diffusive gradient thin film technique

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

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.

Assessment of fungi in soils of sugarcane crops and their potential for production of biomass-degrading enzymes

Soil management practices are konwn to affect the biomass and enzyme activities of microbial soil communities. To assess whether burning of sugarcane prior to harvesting affects the community of soilborne fungi, we collected soil simples in two sites: burned sugarcane culture prior harvesting (BS) and non-burned sugarcane culture (NBS). A total of 75 filamentous fungal isolates were recovered from soils in both sites. Trichoderma was the most prevalent genus in both sites, followed by Fusarium, Cunninghamella and Aspergillus. The Sorensen's index (0.60) suggested a slight difference in fungi associated with both areas, with high number of fungal isolates found on BB soil. The abundance of Trichoderma isolates in NBS soil was higher than BS soil; however, the abundance of Fusarium, Aspergillus and Cunninghamella was higher in the latter type of soil. In addition, fungi isolated from BS soil showed the highest production of xylanase and laccase in comparision with fungi isolated form NBS soil. Our results indicate that the different types of sugarcane harvesting apparently did not interfere with the diversity of fungal communnities as revealed by culture-dependent methods. In addition, our data indicates the potencial of fungi from soils of sugarcane crops to produce relevant enzymes related to biomass conversion.

Modulation of the activity and selectivity of the immobilized lipases by surfactants and solvents

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