Imobilização multipontual covalente de xilanases: seleção de derivados ativos e estabilizados

Pós-graduação em Biotecnologia - IQ

Bioprospecção de bactérias endofíticas como agentes de biocontrole da mancha de Exserohilum turcicum e como promotoras do crescimento de plantas de milho (Zea mays L.)

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Controle da mancha preta dos frutos cítricos em cultivo orgânico e convencional e do bolor verde em pós-colheita

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Estudo da interação da natamicina com membranas fosfolipídicas incorporadas de esteróis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Rhizospheric streptomycetes as potential biocontrol agents of Fusarium and Armillaria pine rot and as PGPR for Pinus taeda

Pinus taeda is one of the main timber trees in Brazil, occupying 1.8 million ha with an annual productivity of 25-30 m(3) ha(-1). Another important species is Araucaria angustifolia, belonging to the fragile Rainforest biome, which for decades has been a major source of timber in Brazil. Some diseases that affect the roots and/or the stem of these trees and cause "damping-off" of the seedlings, with economic and environmental losses for the forest sector, are caused by the plant pathogenic fungi Fusarium sp. or Armillaria sp. This research project intended to isolate actinobacteria from the Araucaria rhizosphere, which present an antagonistic effect against these fungi. After the selection of the best pathogen inhibitors, morphologic characteristics, enzyme production, and their effect on the growth of Pinus taeda were studied. The actinobacteria were tested for their antagonistic capacity against Fusarium sp. in Petri plates with PDA as substrate. The inhibition zone was measured after 3, 5, 7, and 10 days. Of all the isolates tested, only two of them maintained inhibition zones up to 4 mm for 10 days. The inhibition of Armillaria sp. was tested in liquid medium and also in Petri dishes through the evaluation of the number of the fungal rhizomorphs in dual culture with the actinobacteria. It was found that all five isolates were able to inhibit the rhizomorph production, with the best performance of the isolate A43, which was capable of inhibiting both fungi, Fusarium and Armillaria. In a greenhouse experiment, the effect of five isolates on the growth of Pinus taeda seedlings was tested. Plant height, stem diameter, root and shoot dry matter were determined. The Streptomyces isolate A43 doubled plant growth. These results may lead to the development of new technologies in the identification of still unknown bacterial metabolites and new management techniques to control forest plant diseases.

Antagonismo de actinobactérias isoladas de Trachymyrmex frente ao fungo parasita Escovopsis

As formigas da tribo Attini mantêm uma associação com actinobactérias encontradas nos biofilmes presentes no integumento desses insetos. Dentre as actinobactérias associadas às formigas Attini, os gêneros Pseudonocardia e Streptomyces são considerados prevalentes. Estudos indicam que essas bactérias auxiliam na defesa do jardim contra patógenos através da secreção de compostos antimicrobianos. Recentemente foi sugerido que tais bactérias podem ser importantes na defesa do próprio inseto, especialmente contra entomopatógenos. Duas teorias principais foram propostas para explicar as interações entre actinobactérias, formigas Attini e o microfungo parasita Escovopsis. Uma delas defende a coevolução entre esses organismos e a outra rejeita a existência da mesma, argumentando que a interação formiga-actinobactéria é uma simbiose aberta. Neste trabalho, avaliamos a interação entre actinobactérias e Escovopsis em experimentos in vitro. Um total de 14 linhagens de actinobactérias, compreendendo 12 de Pseudonocardia, uma de Streptomyces e uma de Actinoplanes, isoladas de Trachymyrmex (um gênero de formiga Attini não cortadeira) foram testadas frente a quatro linhagens de Escovopsis. Os Escovopsis foram isolados tanto de jardins de fungos de formigas Trachymyrmex quanto de cortadeiras de folhas (Acromyrmex e Atta). O crescimento micelial de Escovopsis (em cm²) foi medido na presença e na ausência (controle) das actinobactérias. Os resultados demonstraram que todas as actinobactérias testadas inibiram Escovopsis, mas as taxas de inibição foram variáveis dependendo da linhagem da actinobactéria (p < 0,05). A linhagem de Streptomyces foi a mais efetiva, corroborando dados da literatura que indicam que outros micro-organismos, além de Pseudonocardia, também apresentam ação inibitória frente à Escovopsis... (Resumo completo, clicar acesso eletrônico abaixo)

Análise e expressão de genes de PKS II e de carboidrases provenientes de bibliotecas metagenômicas

Pós-graduação em Microbiologia Agropecuária - FCAV

Chitin-degrading enzymes from an actinomycete ectosymbiont of Acromyrmex subterraneus brunneus (Hymenoptera: Formicidae)

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

The ribB FMN riboswitch from Escherichia coli operates at the transcriptional and translational level and regulates riboflavin biosynthesis

FMN riboswitches are genetic elements that, in many bacteria, control genes responsible for biosynthesis and/or transport of riboflavin (vitamin B2 ). We report that the Escherichia coli ribB FMN riboswitch controls expression of the essential gene ribB coding for the riboflavin biosynthetic enzyme 3,4-dihydroxy-2-butanone-4-phosphate synthase (RibB; EC 4.1.99.12). Our data show that the E. coli ribB FMN riboswitch is unusual because it operates at the transcriptional and also at the translational level. Expression of ribB is negatively affected by FMN and by the FMN analog roseoflavin mononucleotide, which is synthesized enzymatically from roseoflavin and ATP. Consequently, in addition to flavoenzymes, the E. coli ribB FMN riboswitch constitutes a target for the antibiotic roseoflavin produced by Streptomyces davawensis.

Análise estrutural de halogenases encontradas em clusters gênicos da biossíntese de antibióticos glicopeptídicos

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

Estudo acelerado de estabilidade de hiclato de doxiciclina em comprimidos

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

Estudo acelerado de estabilidade de hiclato de doxiciclina em comprimidos

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

Analysis of the Organic Hydroperoxide Response of Chromobacterium violaceum Reveals That OhrR Is a Cys-Based Redox Sensor Regulated by Thioredoxin

Organic hydroperoxides are oxidants generated during bacterial-host interactions. Here, we demonstrate that the peroxidase OhrA and its negative regulator OhrR comprise a major pathway for sensing and detoxifying organic hydroperoxides in the opportunistic pathogen Chromobacterium violaceum. Initially, we found that an ohrA mutant was hypersensitive to organic hydroperoxides and that it displayed a low efficiency for decomposing these molecules. Expression of ohrA and ohrR was specifically induced by organic hydroperoxides. These genes were expressed as monocistronic transcripts and also as a bicistronic ohrR-ohrA mRNA, generating the abundantly detected ohrA mRNA and the barely detected ohrR transcript. The bicistronic transcript appears to be processed. OhrR repressed both the ohrA and ohrR genes by binding directly to inverted repeat sequences within their promoters in a redox-dependent manner. Site-directed mutagenesis of each of the four OhrR cysteine residues indicated that the conserved Cys21 is critical to organic hydroperoxide sensing, whereas Cys126 is required for disulfide bond formation. Taken together, these phenotypic, genetic and biochemical data indicate that the response of C. violaceum to organic hydroperoxides is mediated by OhrA and OhrR. Finally, we demonstrated that oxidized OhrR, inactivated by intermolecular disulfide bond formation, is specifically regenerated via thiol-disulfide exchange by thioredoxin (but not other thiol reducing agents such as glutaredoxin, glutathione and lipoamide), providing a physiological reducing system for this thiol-based redox switch.

Systematic UPLC-ESI-MS/MS Study on the Occurrence of Staurosporine and Derivatives in Associated Marine Microorganisms from Eudistoma vannamei

Former bioactivity-guided analysis of the marine invertebrate Eudistoma vannamei led to the isolation of staurosporine derivatives, which revealed strong cytotoxic activity against several human cancer cell lines. The occurrence of such alkaloids in E. vannamei may be correlated to the presence of associated biota, such as Streptomyces bacteria. In agreement to this hypothesis, marine microorganisms associated with E. vannamei were recovered and cultured, leading to a total of 84 isolated bacterial strains. Gas phase fragmentation reactions of staurosporine and derivatives were systematically studied and the analyzed results further supported by computational chemistry studies. The resulting fragment patterns were used to search for the presence of different derivatives in extracts of isolated microorganisms, thereby using LC-MS/MS analysis in MRM mode. These results evidenced that one isolated Streptomyces sp. was able to generate staurosporine, while none of the hydroxy-7-oxo derivatives were detected. Finally, significant cytotoxic activity against human cancer lines was observed for one of the extracts.

Systematic UPLC-ESI-MS/MS study on the occurrence of staurosporine and derivatives in associated marine microorganisms from Eudistoma vannamei

Former bioactivity-guided analysis of the marine invertebrate Eudistoma vannamei led to the isolation of staurosporine derivatives, which revealed strong cytotoxic activity against several human cancer cell lines. The occurrence of such alkaloids in E. vannamei may be correlated to the presence of associated biota, such as Streptomyces bacteria. In agreement to this hypothesis, marine microorganisms associated with E. vannamei were recovered and cultured, leading to a total of 84 isolated bacterial strains. Gas phase fragmentation reactions of staurosporine and derivatives were systematically studied and the analyzed results further supported by computational chemistry studies. The resulting fragment patterns were used to search for the presence of different derivatives in extracts of isolated microorganisms, thereby using LC-MS/MS analysis in MRM mode. These results evidenced that one isolated Streptomyces sp. was able to generate staurosporine, while none of the hydroxy-7-oxo derivatives were detected. Finally, significant cytotoxic activity against human cancer lines was observed for one of the extracts.