Giant Amazonian fish pirarucu (Arapaima gigas): Its viscera as a source of thermostable trypsin

A trypsin was purified from pyloric caeca of pirarucu (Arapaima gigas). the effect of metal ions and protease inhibitors on its activity and its physicochemical and kinetic properties, as well its N-terminal sequence, were determined. A single band (28.0 kDa) was observed by SDS-PAGE. Optimum pH and temperature were 9.0 and 65 degrees C, respectively. the enzyme was stable after incubation for 30 min in a wide pH range (6.0-11.5) and at 55 degrees C. the kinetic parameters K-m, k(cat) and k(cat)/K-m were 0.47 +/- 0.042 mM, 1.33 s(-1) and 2.82 s(-1) mM(-1), respectively, using BApNA as substrate. This activity was shown to be very sensitive to some metal ions, such as Fe2+, Hg2+, Zn2+, Al3+, Pb2+, and was highly inhibited by trypsin inhibitors. the trypsin N-terminal sequence IVGGYECPRNSVPYQ was found. the features of this alkaline peptidase suggest that it may have potential for industrial applications (e.g. food and detergent industries). (C) 2012 Elsevier B.V. All rights reserved.Financiadora de Estudos e Projetos (FINEP/RECAR-CINE)Ministerio da Aquicultura e Pesca (MAP)Empresa brasileira de pesquisa agropecuaria (Embrapa)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundacao de Apoio a Ciencia e Tecnologia do Estado de Pernambuco (FACEPE)Petroleo do Brasil S/A (PETROBRAS)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Univ Fed Pernambuco, Lab Enzimol LABENZ, Dept Bioquim, BR-50670420 Recife, PE, BrazilUniv Fed Pernambuco, LIKA, BR-50670420 Recife, PE, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, BR-04044020 São Paulo, BrazilWeb of Scienc

Metal-sensitive and thermostable trypsin from the crevalle jack (Caranx hippos) pyloric caeca: purification and characterization

Background: Over the past decades, the economic development and world population growth has led to increased for food demand. Increasing the fish production is considered one of the alternatives to meet the increased food demand, but the processing of fish leads to by-products such as skin, bones and viscera, a source of environmental contamination. Fish viscera have been reported as an important source of digestive proteases with interesting characteristics for biotechnological processes. Thus, the aim of this study was to purify and to characterize a trypsin from the processing by-products of crevalle jack (Caranx hippos) fish.Results: A 27.5 kDa trypsin with N-terminal amino acid sequence IVGGFECTPHVFAYQ was easily purified from the pyloric caeca of the crevalle jack. Its physicochemical and kinetic properties were evaluated using N-alpha-benzoyl-(DL)-arginine-p-nitroanilide (BApNA) as substrate. in addition, the effects of various metal ions and specific protease inhibitors on trypsin activity were determined. Optimum pH and temperature were 8.0 and 50 degrees C, respectively. After incubation at 50 degrees C for 30 min the enzyme lost only 20% of its activity. K-m, k(cat), and k(cat)/K-m values using BApNA as substrate were 0.689 mM, 6.9 s(-1), and 10 s(-1) mM(-1), respectively. High inhibition of trypsin activity was observed after incubation with Cd2+, Al3+, Zn2+, Cu2+, Pb2+, and Hg2+ at 1 mM, revealing high sensitivity of the enzyme to metal ions.Conclusions: Extraction of a thermostable trypsin from by-products of the fishery industry confirms the potential of these materials as an alternative source of these biomolecules. Furthermore, the results suggest that this trypsin-like enzyme presents interesting biotechnological properties for industrial applications.Financiadora de Estudos e Projetos (FINEP/RECARCINE)Petroleo do Brasil S/A (PETROBRAS)Secretaria Especial de Aquicultura e Pesca (SEAP/PR)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundacao de Apoio a Ciencia e Tecnologia do Estado de Pernambuco (FACEPE)Univ Fed Pernambuco, Lab Enzimol LABENZ, Dept Bioquim CCB, BR-50670910 Recife, PE, BrazilUniv Fed Pernambuco, LIKA, BR-50670910 Recife, PE, BrazilUniversidade Federal de São Paulo, Dept Biofis, Escola Paulista Med, BR-04044020 São Paulo, BrazilUniv Fed Pernambuco, Lab Glicoprot, Dept Bioquim CCB, BR-50670910 Recife, PE, BrazilUniversidade Federal de São Paulo, Dept Biofis, Escola Paulista Med, BR-04044020 São Paulo, BrazilWeb of Scienc

Aspectos do queijo de coalho com ênfase na importância das Boas Práticas de Fabricação no sistema de produção / Aspects of coalho type cheese with emphasis on the importance of Good Manufacturing Practices in the production system

O queijo de coalho é um produto típico do Nordeste e bastante consumido pela população regional, exercendo influência de âmbito cultural, social e econômico. Este alimento tem a maioria da sua produção realizada de forma artesanal em áreas rurais de pequeno porte ou de agricultura de base familiar, sendo sua imagem atrelada a um produto de baixa qualidade microbiológica devido a sua produção ser comumente baseada na utilização de matéria-prima in natura e na ausência de práticas adequadas de fabricação. Vários estudos indicam a presença de contaminação por bactérias no queijo de coalho, tais como: Salmonella spp., Staphylococcus aureus e Escherichia coli, podendo estes microrganismos causar riscos para a saúde humana. Sendo assim, a aplicação de Boas Práticas de Fabricação (BPFs) é de fundamental importância para manter a qualidade do queijo de coalho mantendo-o seguro quanto ao padrão de inocuidade regulamentado. Dessa forma, objetivou-se com este estudo identificar evidências disponíveis acerca dos aspectos do queijo de coalho com ênfase na importância das Boas Práticas de Fabricação no sistema de produção. Para tanto, realizou-se um levantamento bibliográfico descritivo e qualitativo utilizando: legislações, artigos completos, monografias, dissertações, teses e site institucional publicadas nos idiomas, português, inglês e espanhol, com dimensão temporal entre 1996 e 2016. Verificou-se que a maior parte da produção do queijo de coalho é realizada de forma artesanal ou por empreendimentos de médio e pequeno porte, ambientes estes, que possuem importante resistência dos produtores quanto a procedimentos adequados e legítimos de elaboração do alimento, tornando consequentemente, grande parte do queijo de coalho susceptível a contaminação microbiológica devido à ausência de práticas higiênico-sanitárias nestes ambientes de produção, o que indica a necessidade da aplicação das BPFs a fim de, adequar o sistema produtivo deste derivado lácteo, visto que, diversos estudos demonstraram que as realizações dos requisitos das BPFs trazem uma sensibilização, aperfeiçoamento e conhecimento dos proprietários de queijarias e seus colaboradores, além de melhorar as condições de produção, processamento e comercialização, agregando valor substancial ao laticínio além de, garantir um alimento seguro para o consumo humano no que tange aos aspectos nutricionais e microbiológicos. 

LIBERDADE ACADÊMICA EM TEMPOS DIFÍCEIS: DIÁLOGOS BRASIL E ESTADOS UNIDOS

This paper intends to discuss the academic freedom, its role and its possible limits and borders, from the paradigm of the Democratic Constitutional State, as configured in our constitutional text of October 1988. The discussion focuses on the relationship between the critical and investigative role expected of the Universities and of its members in relation to the so-called legislative proposals of the Escola Sem Partido movement. Hence, it explores if these proposals do not offend the constitutional freedom of professors, students, technicians and researchers, in addition to the higher education institutions themselves, when subjects that gravitate around their roles and responsibilities are taken into account. For this purpose, keeping in mind our own constitutional context, we decided to use Comparative Law, focusing on decisions of the Supreme Court of the United States that deal with the same issue addressed here. The analysis indicates that academic freedom has an essential and constitutionally recognized role in the Constitutional Democracy.O artigo busca trabalhar a liberdade acadêmica, seu papel e seus possíveis limites e fronteiras, a partir do paradigma do Estado Democrático de Direito, como configurado em nosso texto constitucional de outubro de 1988. O enfoque se dá na relação entre o papel crítico e investigativo esperado das Universidades e de seus membros, diante das chamadas propostas legislativas do movimento Escola Sem Partido. Isto é, perguntamos se estas propostas não ofenderiam a liberdade constitucional de professores, alunos, técnicos e pesquisadores, além das próprias instituições de ensino superior, quanto a assuntos que gravitam em torno de suas funções e responsabilidades. Para este fim, sem olvidarmos de nosso próprio contexto constitucional, decidimos dialogar com o direito comparado, principalmente com o cenário estadunidense e certas decisões da Suprema Corte dos Estados Unidos que lidam com a mesma temática aqui abordada. Conclui-se que a liberdade acadêmica tem um papel essencial, constitucionalmente reconhecido, no Estado Democrático de Direito.

Maestro Duda : a vida e a obra de um compositor da terra do frevo

Orientador: Ricardo GoldembergDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de ArtesResumo: A falta de divulgação de alguns compositores brasileiros, e a inexistência de uma catalogação adequada de suas obras, fazem com que uma grande parte da memória musical brasileira seja perdida. O presente trabalho fornece dados biográficos e detalhes sobre a trajetória musical do compositor pernambucano José Ursicino da Silva, maestro Duda, bem como alguns aspectos relevantes de seu estilo musical e a catalogação de sua obra. O catálogo foi estruturado a partir de levantamento realizado em diversas instituições musicais das cidades de Recife, João Pessoa e Natal, e arquivos particularesAbstract: This dissertation is about the life and work of José Ursicino da Silva (maestro Duda), a Brazilian composer who was bom in the northeastem state of Pemambuco. Ris works includes compositions for trumpet with piano accompaniment, brass quintet, symphonic orchestra and others.The work offers biographical and musical informations about the composer as well as important aspects about his musical style. Also, a catalogue of his compositions, based on a survey done in musical institutions ofRecife, Joao Pessoa and Natal, as well in private archives is presented.MestradoMestre em Músic

Cytotoxic Compounds Derived from Marine Sponges. A Review (2010–2012)

Abstract: This extensive review covers research published between 2010 and 2012 regarding new compounds derived from marine sponges, including 62 species from 60 genera belonging to 33 families and 13 orders of the Demospongia class (Porifera). The emphasis is on the cytotoxic activity that bioactive metabolites from sponges may have on cancer cell lines. At least 197 novel chemical structures from 337 compounds isolated have been found to support this work. Details on the source and taxonomy of the sponges, their geographical occurrence, and a range of chemical structures are presented. The compounds discovered from the reviewed marine sponges fall into mainly four chemical classes: terpenoids (41.9%), alkaloids (26.2%), macrolides (8.9%) and peptides (6.3%) which, along with polyketides, sterols, and others show a range of biological activities. The key sponge orders studied in the reviewed research were Dictyoceratida, Haplosclerida, Tetractinellida, Poecilosclerida, and Agelasida. Petrosia, Haliclona (Haplosclerida), Rhabdastrella (Tetractinellida), Coscinoderma and Hyppospongia (Dictyioceratida), were found to be the most promising genera because of their capacity for producing new bioactive compounds. Several of the new compounds and their synthetic analogues have shown in vitro cytotoxic and pro-apoptotic activities against various tumor/cancer cell lines, and some of them will undergo further in vivo evaluation

Digestive peptidases and proteinases in the midgut gland of the pink shrimp Farfantepenaeus paulensis (Crustacea, Decapoda, Penaeidae)

Proteases from the midgut gland of the Farfantepenaeus paulensis juveniles were assessed. Enzyme activity was determined using protease substrates and inhibitors. The effect of pH, temperature and calcium on proteolytic activity was assayed. Caseinolytic activity was analysed in substrate-sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Trypsin, chymotrypsin and leucine aminopeptidase activity was detected. Proteolytic activity was strongly inhibited by the specific trypsin inhibitors. Tosyl-phenylalanine chloromethyl ketone inhibited 59.3% of chymotrypsin activity. The greatest trypsin-like activity occurred at pH 8.0 and 45 degrees C. Chymotrypsin-like activity reached maximal values at alkaline pH (7.2-9.0) and 55 degrees C. CaCl(2) did not increase trypsin-like activity, but rather inhibited it at concentrations of 30 (20%), 50 (30%) and 100 mM (50%). The substrate-SDS-PAGE zymogram revealed eight proteinase bands. Two possibly thermal-resistant (85 degrees C, 30 min) chymotrypsin isoforms were found, which were inhibited by phenyl-methyl-sulphonyl-fluoride. Aminopeptidase activity of enzyme extracts (Arg, Leu, Lys, Phe and Val) and the recommended concentrations of these essential amino acids in penaeid shrimp diets were positively correlated (P < 0.05). Beause protein digestion involves the combined action of different enzymes, adequate knowledge of shrimp digestion and enzyme characteristics is required for the assessment of the digestive potential of different feed sources and development of in vitro digestibility protocols.FINEP/RECARCINEFinanciadora de Estudos e Projetos (FINEP)SEAP/PRSEAP/PRConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)CNPqFACEPEFACEPEPETROBRASPETROBRASFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP[05/50578-2]FAPESP[07/07051-9]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)CNPq/SEAP[504031/2003-1]CNPq/SEAPConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)CNPq fellowship[308444/2006-0]National Research System (Brazil)National Research System (Brazil