Molecular fragmentation of wheat-germ agglutinin induced by food irradiation reduces its allergenicity in sensitised mice

WGA, an agglutinin from wheat germ which is largely responsible for many of wheat's allergies, was used as a model to investigate the action of ionising radiation on WGA's anti-nutritive effects in sensitised mice. Based on the molecular structure, the present study also examined the structural modification of WGA in relation to the range of dose. Structural integrity was monitored using HPLC, fluorescence spectrometry and circular dichroism. Results showed a loss of intrinsic activity and the formation of insoluble amorphous aggregates with a lack of native conformational structures after irradiation. Current findings suggest that the allergenic epitopes of WGA became less active and antigenic after high-dose radiation. the reduction of cytokines typical of allergic reactions, with decreased lymphocytic infiltrate, was observed in the gut of mice given irradiated versus native WGA. Food irradiation proved effective and safe in combating immunological and allergic effects of WGA. (C) 2011 Elsevier B.V. All rights reserved.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Ministerio da Ciencia e Tecnologia (Brazilian)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundacao de Amparo a Ciencia e Tecnologia do Estado de Pernambuco (FACEPE)Univ Fed Pernambuco, Dept Bioquim, Recife, PE, BrazilUniv Fed Pernambuco, Dept Histol & Embriol, Recife, PE, BrazilUniv Fed Pernambuco, Dept Biofis & Radiobiol, Recife, PE, BrazilUniv Fed Pernambuco, Dept Antibiot, Recife, PE, BrazilUniv Estadual Oeste Parana, Ctr Engn & Ciencias Exatas, Toledo, BrazilUniversidade Federal de São Paulo, Dept Bioquim, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Bioquim, São Paulo, BrazilWeb of Scienc

Interaction of proteinase inhibitors with phospholipid vesicles is modulated by pH

rBbKI and rBbCI, plant recombinant inhibitors from Bauhinia bauhinioides, and BpuTI from Bauhinia purpurea seeds distinctly and specifically block proteolytic enzymes. the secondary structures of those inhibitors were compared and their interactions with phospholipid vesicles were evaluated by the release of calcein and by intrinsic fluorescence of tryptophan residues. the results show that rBbKI, rBbCI and BpuTI are able to interact with phospholipd vesicles and induce membrane permeabilization in a concentration- and pH-dependent manner. the leakage was rapid and extensive at pH 4.5, but at physiological pH, no calcein release was observed. These results may suggest that upon inflammation or microorganism invasion accompanied by lowering of pH, appropriate conditions may occur for the inhibitors to interact with cell membrane and act on specific proteolytic enzyme. (C) 2010 Elsevier B.V. All rights reserved.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Escola Paulista Med, Dept Bioquim, BR-04044020 São Paulo, BrazilUniv Estadual Oeste Parana, Ctr Engn & Ciencias Exatas, BR-85903000 Toledo, PR, BrazilUniv Coimbra, Fac Ciencias & Tecnol, Ctr Neurociencias & Biol Celular, Dept Ciencias Vida, P-3001401 Coimbra, PortugalUniversidade Federal de São Paulo, Escola Paulista Med, Dept Bioquim, BR-04044020 São Paulo, BrazilWeb of Scienc

The defensive functions of plant inhibitors are not restricted to insect enzyme inhibition

Three plant proteinase inhibitors BbKI (kallikrein inhibitor) and BbCI (cruzipain inhibitor) from Bauhinia bouhinioides, and a BrTI (trypsin inhibitor) from B. rufa, were examined for other effects in Callosobruchus maculatus development; of these only BrTI affected bruchid emergence. BrTI and BbKI share 81% identities in their primary sequences and the major differences between them are the regions comprising the RGD and RGE motifs in BrTI. These sequences were shown to be essential for BrTI insecticidal activity, since a modified BbKI [that is a recombinant form (BbKIm) with some amino acid residues replaced by those found in BrTI sequence] also strongly inhibited insect development. By using synthetic peptides related to the BrTI sequence, YLEAPVARGDGGLA-NH(2) (RGE) and IVYYPDRGETGL-NH(2) (RGE), it was found that the peptide with an RGE sequence was able to block normal development of C. maculatus larvae (ED(50) 0.16% and LD(50) 0.09%), this being even more effective than the native protein. (C) 2009 Elsevier B.V. All rights reserved.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAP/FADA (UNIFESP)Universidade Federal de São Paulo, Escola Paulista Med, Dept Bioquim, BR-04044020 São Paulo, BrazilUniv Fed Mato Grosso, Dept Nat Sci, BR-79603011 Tres Lagoas, MS, BrazilUniv Estadual Fluminense Darcy Ribeiro, Lab Prot & Peptide Biochem, CBB, BR-28015620 Campos Dos Goytacazes, RJ, BrazilUniversidade Federal de São Paulo, Dept Biophys, BR-04044020 São Paulo, BrazilUniv São Paulo, Inst Fis Sao Carlos, Sao Carlos, SP, BrazilUniv Estadual Oeste Parana, Ctr Engn & Ciencias Exatas, BR-85903000 Toledo, PR, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Bioquim, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biophys, BR-04044020 São Paulo, BrazilWeb of Scienc

Immobilized Cratylia mollis lectin: An affinity matrix to purify a soybean (Glycine max) seed protein with in vitro platelet antiaggregation and anticoagulant activities

Lectins, proteins which recognize selectively carbohydrates, have been used as affinity chromatography to purify glycoproteins. Cratylia mollis seed lectin (Cramoll 1,4), of mannose/glucose binding class, immobilized on Sepharose CL-4B, was used as affinity matrix. This paper describes the purification by Cramoll 1,4-Sepharose matrix, and characterization of an anti-platelet and anticoagulant soybean (Glycine max) protein, ApcSP, and its in vitro platelet antiaggregation and anticoagulant activities. SDS-PAGE of ApcSP purified under reducing condition revealed a single glycosilated band of 51 kDa. the N-terminal 10-residue sequence of ApcSP is EGQFGPMIQS, distinct to other soybean seed proteins, such as peroxidase, lectin, 2S albumin and trypsin inhibitor. Deconvolution of CD spectrum indicated 35% alpha-helix, 17% beta-strand, 22% turn and 26% unordered structure; ApcSP fluorescence spectrum showed a maximum emission around 339 nm. the hemostatic parameters revealed inhibition of collagen (p<0.001), thrombin (p<0.05) and ADP (p<0.001)-induced platelet aggregation in the presence of ApcSP (2 mu M), in relation to positive control. the soy protein prolonged the blood coagulation time (activated partial thromboplastin time, more affected, and prothrombin time). the results indicated that immobilized Cramoll 1,4 lectin has the potential to isolate soybean glycoproteins and ApcSP may be important for anti-thrombotic and anticoagulant therapy. (C) 2010 Elsevier B.V. All rights reserved.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundacao de Amparo a Ciencia e Tecnologia do Estado de Pernambuco (FACEPE)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Univ Fed Pernambuco, Dept Bioquim, BR-50670420 Recife, PE, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Bioquim, BR-04044020 São Paulo, BrazilUniv Estadual Oeste Parana, Ctr Engn & Ciencias Exatas, BR-85903000 Toledo, PR, BrazilUniv Fed Rio de Janeiro, Inst Bioquim Med, BR-21941590 Rio de Janeiro, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Bioquim, BR-04044020 São Paulo, BrazilWeb of Scienc

High doses of gamma radiation suppress allergic effect induced by food lectin

One of the most promising areas for the development of functional foods lies in the development of effective methods to reduce or eliminate food allergenicity, but few reports have summarized information concerning the progress made with food irradiation. in this study, we investigated the relationship between allergenicity and molecular structure of a food allergen after gamma irradiation and evaluate the profile of the allergic response to irradiated allergens. Cramoll, a lectin isolated from a bean and used as a food allergen, was irradiated and the possible structural changes were accompanied by spectrofluorimetry, circular dichroism and microcalorimetry. Subsequently, sensitized animals subjected to intragastric administration of non-irradiated and irradiated Cramoll were treated for 7 days. Then, body weight, leukocytes, cytokine profiles and histological parameters were also determined. Cramoll showed complete inhibition of intrinsic activity after high radiation doses. Changes in fluorescence and CD spectra with a simultaneous collapse of the tertiary structure followed by a pronounced decrease of native secondary structure were observed after irradiation. After oral challenge, sensitized mice demonstrate an association between Cramoll intake, body weight loss, eosinophilia, lymphocytic infiltrate in the gut and Eotaxin secretion. Irradiation significantly reduces, according to the dose, the effects observed by non-irradiated food allergens. We confirm that high-dose radiation may render protein food allergens innocuous by irreversibly compromising their molecular structure. (C) 2012 Elsevier B.V. All rights reserved.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Ministerio da Ciencia e Tecnologia (Brazilian)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundacao de Amparo a Ciencia e Tecnologia do estado de Pernambuco (FACEPE)Univ Fed Campina Grande, Ctr Saude & Tecnol Rural, BR-58700970 Patos de Minas, Paraiba, BrazilUniv Fed Pernambuco, Dept Bioquim, Recife, PE, BrazilUniv Fed Pernambuco, Dept Fisiol & Farmacol, Recife, PE, BrazilUniv Fed Pernambuco, Dept Antibiot, Recife, PE, BrazilUniv Fed Pernambuco, Dept Histol & Embriol, Recife, PE, BrazilUniv Fed Pernambuco, Dept Biofis & Radiobiol, Recife, PE, BrazilUniv Estadual Oeste Parana, Ctr Engn & Ciencias Exatas, Toledo, BrazilUniversidade Federal de São Paulo, Dept Bioquim, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, São Paulo, BrazilUniv São Paulo, Dept Bioquim, Inst Quim, BR-05508 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Bioquim, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, São Paulo, BrazilWeb of Scienc

Bioenergia: desenvolvimento, pesquisa e inovação

Com 27 trabalhos produzidos por pesquisadores do Instituto de Pesquisa em Bioenergia (Bioen), da Unesp, este livro oferece uma ampla visão sobre as áreas que compõem o segmento. Seu principal objetivo é contribuir para melhorar a compreensão dos vários aspectos da bioenergia, em especial no Brasil, que figura entre os países com maior nível de desenvolvimento tecnológico no setor. Os artigos abordam uma série abrangente de questões relacionadas à bioenergia, como a construção genética das plantas de cana-de-açúcar visando ao aumento de produtividade, a disseminação de sementes para estimular a propagação de espécies com potencial energético, etapas de produção de bioenergia, usos do combustível e seus efeitos nos diversos tipos de motores. Agrupados por assunto, os textos estão distribuídos em cinco partes: Biomassa para bioenergia; Produção de biocombustíveis; Utilização de bioenergia; Biorrefinaria, alcoolquímica e oleoquímica e Sustentabilidade dos biocombustíveis