Author | Waniek, Peter J. | |
Author | Lima, Leila Mendonça | |
Author | Menezes, Gustavo B. | |
Author | Jansen, Ana Maria | |
Author | Araújo, Catarina A. C. | |
Access date | 2017-08-31T11:24:24Z | |
Available date | 2017-08-31T11:24:24Z | |
Document date | 2009 | |
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. | pt_BR |
ISSN | 0307-6962 | pt_BR |
URI | https://www.arca.fiocruz.br/handle/icict/20844 | |
Language | eng | pt_BR |
Publisher | Wiley | pt_BR |
Rights | restricted access | pt_BR |
Subject in Portuguese | Triatoma brasiliensis | pt_BR |
Subject in Portuguese | Digestão | pt_BR |
Subject in Portuguese | Muramidase | pt_BR |
Subject in Portuguese | Intestino médio | pt_BR |
Subject in Portuguese | Simbiontes de insetos | pt_BR |
Title | Recombinant expression and characterization of a lysozyme from the midgut of Triatoma brasiliensis (Hemiptera, Reduviidae) in comparison with intestinal muramidase activity | pt_BR |
Type | Article | pt_BR |
DOI | 10.1111/j.1365-3032.2009.00691.x | |
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. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia de Tripanosomatídeos. Rio de Janeiro, RJ. Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genômica Funcional e Bioinformática. Rio de Janeiro, RJ. Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genômica Funcional e Bioinformática. Rio de Janeiro, RJ. Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia de Tripanosomatídeos. Rio de Janeiro, RJ. Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia de Tripanosomatídeos. Rio de Janeiro, RJ. Brasil. | pt_BR |
Subject | Digestion | pt_BR |
Subject | insect symbionts | pt_BR |
Subject | lysozyme | pt_BR |
Subject | midgut | pt_BR |
Subject | Triatoma brasiliensis | pt_BR |
e-ISSN | 1365-3032 | |
Embargo date | 2030-01-01 | |