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Ig-like Domain in Endoglucanase Cel9A from Alicyclobacillus acidocaldarius Makes Dependent the Enzyme Stability on Calcium

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Abstract

Endoglucanase Cel9A from Alicyclobacillus acidocaldarius (AaCel9A) has an Ig-like domain and the enzyme stability is dependent to calcium. In this study the effect of calcium on the structure and stability of the wild-type enzyme and the truncated form (the wild-type enzyme without Ig-like domain, AaCel9AΔN) was investigated. Fluorescence quenching results indicated that calcium increased and decreased the rigidity of the wild-type and truncated enzymes, respectively. RMSF results indicated that AaCel9A has two flexible regions (regions A and B) and deleting the Ig-like domain increased the truncated enzyme stability by decreasing the flexibility of region B probably through increasing the hydrogen bonds. Calcium contact map analysis showed that deleting the Ig-like domain decreased the calcium contacting residues and their calcium binding affinities, especially, in region B which has a role in calcium binding site in AaCel9A. Metal depletion and activity recovering as well as stability results showed that the structure and stability of the wild-type and truncated enzymes are completely dependent on and independent of calcium, respectively. Finally, one can conclude that the deletion of Ig-like domain makes AaCel9AΔN independent of calcium via decreasing the flexibility of region B through increasing the hydrogen bonds. This suggests a new role for the Ig-like domain which makes AaCel9A structure dependent on calcium.

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Abbreviations

AaCel9A:

Alicyclobacillus acidocaldarius endoglucanase Cel9A

AaCel9AΔN:

AaCel9A without Ig-like domain

CBD:

Carbohydrate binding domain

Ig-like:

Immunoglobulin-like

MD:

Molecular dynamics

RDF:

Radial distribution function

RMSF:

Root-mean-square fluctuation

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Acknowledgements

The authors express their gratitude to the research council of Azarbaijan Shahid Madani University for the financial support during the course of this project.

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Authors and Affiliations

  1. Department of Cellular and Molecular Biology, Faculty of Science, Azarbaijan Shahid Madani University, Tabriz, Iran

    Mohammad Pazhang, Fereshteh S. Younesi, Saeed Najavand & Mehrnaz Haghi

  2. Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran, Tehran, Iran

    Faramarz Mehrnejad

  3. Department of Biotechnology, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, Iran

    Alireza Tarinejad

  4. Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran

    Fatemeh Rashno & Khosro Khajeh

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  1. Mohammad Pazhang
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  2. Fereshteh S. Younesi
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Correspondence to Mohammad Pazhang.

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Pazhang, M., Younesi, F.S., Mehrnejad, F. et al. Ig-like Domain in Endoglucanase Cel9A from Alicyclobacillus acidocaldarius Makes Dependent the Enzyme Stability on Calcium. Mol Biotechnol 60, 698–711 (2018). https://doi.org/10.1007/s12033-018-0105-4

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