Proteomic analysis reveals the damaging role of low redox laccase from Yersinia enterocolitica strain 8081 in the midgut
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ORIGINAL RESEARCH PAPER
Proteomic analysis reveals the damaging role of low redox laccase from Yersinia enterocolitica strain 8081 in the midgut of Helicoverpa armigera Shruti Ahlawat . Deepti Singh . Asha Yadav . Amarjeet Kumar Singh . Jugsharan Singh Virdi . Krishna Kant Sharma
Received: 21 January 2020 / Accepted: 25 May 2020 Ó Springer Nature B.V. 2020
Abstract 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 (Hu¨bner) 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 (Hu¨bner) larvae can be used to study the effect of microbes and their metabolites on the host physiology, anatomy, and survival.
Shruti Ahlawat and Deepti Singh have contributed equally.
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-02925-x) contains supplementary material, which is available to authorized users. S. Ahlawat D. Singh A. Yadav K. K. Sharma (&) Laboratory of Enzymology and Recombinant DNA Technology, Department of Microbiology, Maharshi Dayanand University, Rohtak, Haryana 124001, India e-mail: [email protected]; [email protected] D. Singh Department of Microbiology, Amity University, Jaipur, India
A. K. Singh Centre for Genetic Manipulation of Crop Plants, University of Delhi South Campus, Benito Juarez Road, New Delhi 110021, India J. S. Virdi Microbial Pathogenicity Laboratory, Department of Microbiology, University of Delhi South Campus, Benito Juarez Road, New Delhi 110021, India
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Biotechnol Lett
Graphic abstract
Keywords Laccase Cloning Protein purification Yersinia enterocolitica Helicoverpa armigera Proteome Reactive oxygen species
Introduction Yersinia enterocolitica is a potential human pathogen with extensive serotype diversity, which is further differentiate
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