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3 Biotech

, 8:352 | Cite as

Cloning and characterization of trehalase: a conserved glycosidase from oriental midge, Chironomus ramosus

  • Ekta ShuklaEmail author
  • Leena Thorat
  • Ameya D. Bendre
  • Santosh Jadhav
  • Jayanta K. Pal
  • Bimalendu B. Nath
  • Sushama M. Gaikwad
Short Reports

Abstract

Insect trehalase is a multiferous enzyme, crucial for normal physiological functions as well as under stress conditions. In this report, we present a fundamental study of the trehalase gene segment (1587 bp) from Chironomus ramosus (CrTre) encoding for 529 amino acids, using appropriate bioinformatics tools. C. ramosus, a tropical midge is an emerging animal model to investigate the consequences of environmental stresses. We observed that CrTre belongs to GH family 37 in the CAZy database and possess 57–92% identity to dipteran trehalases. In silico characterization provided information regarding the structural, functional and evolutionary aspects of midge trehalase. In the phylogenetic tree, CrTre clustered with the soluble dipteran trehalases. Moreover, domain functional characterization of the deduced protein sequence by InterProScan (IPR001661), ProSite (PS00927 and PS00928) and Pfam (PF01204) indicated presence of highly conserved signature motifs which are important for the identification of trehalase superfamily. Furthermore, the instability index of CrTre was predicted to be < 40 suggesting its in vivo stability while, the high aliphatic index indicated towards its thermal stability (index value 71–81). The modelled 3D tertiary structure of CrTre depicts a (α/α)6 barrel toroidal core. The catalytic domain of the enzyme comprised Glu424 and Asp226 as the putative active site residues. Interestingly, the conserved motifs were observed to be formed by the flexible loopy regions in the tertiary structure. This study revealed essential sequence features of the midge trehalase and offers better insights into the structural aspects of this enzyme which can be correlated with its function.

Keywords

Trehalose Midges Conserved motifs Homology modelling Secondary structure 

Abbreviations

CrTre

C. ramosus trehalase

dNTP

Deoxy nucleotide triphosphate

MEGA

Molecular evolutionary genetics analysis

min

Minutes

NCBI

National Centre for BIotechnology

PCR

Polymerase chain reaction

PDB

Protein Data Bank

pI

Isoelectric point

Notes

Acknowledgements

This work is supported by funding received from DRDP programme, Department of Biotechnology, SPPU and DST-PURSE grants to JKP and from partial funding received from DST-PURSE and BCUD-UoP grants to BBN. The authors thank Dr. Varsha Bhatia, Gennova Biopharmaceuticals Ltd. and Mr. Ejaj Pathan, CSIR-NCL for their valuable suggestions and timely help. ES and ADB acknowledge University Grants Commission, New Delhi, for Senior Research Fellowships. LT is grateful for financial support received from the University Grants Commission-DS Kothari Postdoctoral Fellowship (UGC-DSK-PDF) and from DBT Bio-CARe Women Scientist Scheme, New Delhi, India.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest in the publication.

Supplementary material

13205_2018_1376_MOESM1_ESM.docx (5.3 mb)
Supplementary material 1 (DOCX 5461 KB)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Ekta Shukla
    • 1
    • 2
    • 5
    Email author
  • Leena Thorat
    • 3
  • Ameya D. Bendre
    • 1
    • 2
  • Santosh Jadhav
    • 4
    • 6
  • Jayanta K. Pal
    • 4
    • 7
  • Bimalendu B. Nath
    • 3
  • Sushama M. Gaikwad
    • 1
    • 2
  1. 1.Division of Biochemical SciencesCSIR-National Chemical LaboratoryPuneIndia
  2. 2.Academy of Scientific and Innovative Research (AcSIR)PuneIndia
  3. 3.Stress Biology Research Laboratory, Department of ZoologyS.P. Pune UniversityPuneIndia
  4. 4.Cell and Molecular Biology Laboratory, Department of BiotechnologyS.P. Pune UniversityPuneIndia
  5. 5.National Centre for Cell SciencePuneIndia
  6. 6.BAIF Central Research StationPuneIndia
  7. 7.Dr. D.Y. Patil Biotechnology and Bioinformatics InstitutePuneIndia

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