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Biotechnol Lett ; 42(11): 2189-2210, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32472187

RESUMO

OBJECTIVE: Earlier, we have found that the enteropathogenic Yersinia enterocolitica have evolved the survival mechanisms that regulate the expression of laccase-encoding genes in the gut. The present study aims to characterize the purified recombinant laccase from Y. enterocolitica strain 8081 biovar 1B and understand its effect on the midgut of cotton bollworm, Helicoverpa armigera (Hübner) larvae. RESULTS: The recombinant laccase protein showed high purity fold and low molecular mass (~ 43 kDa). H. armigera larvae fed with laccase protein showed a significant decrease in body weight and damage in the midgut. Further, transmission electron microscopy (TEM) studies revealed the negative effect of laccase protein on trachea, malpighian tubules, and villi of the insect. The proteome comparison between control and laccase-fed larvae of cotton bollworm showed significant expression of proteolytic enzymes, oxidoreductases, cytoskeletal proteins, ribosomal proteins; and proteins for citrate (TCA cycle) cycle, glycolysis, stress response, cell redox homeostasis, xenobiotic degradation, and insect defence. Moreover, it also resulted in the reduction of antioxidants, increased melanization (insect innate immune response), and enhanced free radical generation. CONCLUSIONS: All these data collectively suggest that H. armigera (Hübner) larvae can be used to study the effect of microbes and their metabolites on the host physiology, anatomy, and survival.


Assuntos
Proteínas de Insetos/metabolismo , Lacase/toxicidade , Lepidópteros/crescimento & desenvolvimento , Proteômica/métodos , Yersinia enterocolitica/enzimologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/toxicidade , Peso Corporal/efeitos dos fármacos , Clonagem Molecular , Trato Gastrointestinal/diagnóstico por imagem , Trato Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/patologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Proteínas de Insetos/efeitos dos fármacos , Lacase/genética , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Lepidópteros/efeitos dos fármacos , Microscopia Eletrônica de Transmissão , Peso Molecular , Yersinia enterocolitica/genética
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