Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/20844
Title: Recombinant expression and characterization of a lysozyme from the midgut of Triatoma brasiliensis (Hemiptera, Reduviidae) in comparison with intestinal muramidase activity
Authors: Waniek, Peter J.
Lima, Leila Mendonça
Menezes, Gustavo B.
Jansen, Ana Maria
Araújo, Catarina A. C.
Affilliation: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia de Tripanosomatídeos. Rio de Janeiro, RJ. Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genômica Funcional e Bioinformática. Rio de Janeiro, RJ. Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genômica Funcional e Bioinformática. Rio de Janeiro, RJ. Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia de Tripanosomatídeos. Rio de Janeiro, RJ. Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia de Tripanosomatídeos. Rio de Janeiro, RJ. Brasil.
Abstract: Insect c-type lysozymes are antibacterial proteins that are synthesized in different organs with high activity against Gram-positive bacteria. Because lysozymes possess muramidase activity, they also play an important role in the digestion of bacteria in Diptera. Triatomines express lysozyme-encoding genes constitutively in the anterior region (cardia and stomach) of the midgut and the fat body after injection of bacteria into the haemocoel. The present study describes the overexpression of the Triatoma brasiliensis lysozyme 1 (lys1) in Escherichia coli. Recombinant T. brasiliensis Lys1 (TbLys1) is purified after solubilization of the inclusion bodies. The protein refolds successfully, showing muramidase activity against Micrococcus lysodeikticus lyophilized cells, after enterokinase cleavage of its thioredoxin fusion protein. In in-gel zymograms and turbidimetric liquid assays TbLys1 is broadly active under alkaline and acid conditions, indicating a possible digestive function in the two physiologically different midgut regions of the bug: the stomach and small intestine. Muramidase activity is shown in the stomach and small intestine content of unfed bugs and bugs at different days after feeding, respectively. Western blot analysis identifies TbLys1 as lysozyme.
Keywords: Digestion
insect symbionts
lysozyme
midgut
Triatoma brasiliensis
keywords: Triatoma brasiliensis
Digestão
Muramidase
Intestino médio
Simbiontes de insetos
Issue Date: 2009
Publisher: Wiley
Citation: WANIEK, Peter J. et al. Recombinant expression and characterization of a lysozyme from the midgut of Triatoma brasiliensis (Hemiptera, Reduviidae) in comparison with intestinal muramidase activity. Physiological Entomology, v. 34, p.309–317, 2009.
DOI: 10.1111/j.1365-3032.2009.00691.x
ISSN: 0307-6962
Copyright: restricted access
Appears in Collections:IOC - Artigos de Periódicos

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