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A Novel Mini Protein Design of Haloalkane Dehalogenase

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Abstract

The application of native enzymes may not be economical owing to the stability factor. A smaller protein molecule may be less susceptible to external stresses. Haloalkane dehalogenases (HLDs) that act on toxic haloalkanes may be incorporated as bioreceptors to detect haloalkane contaminants. Therefore, this study aims to develop mini proteins of HLD as an alternative bioreceptor which was able to withstand extreme conditions. Initially, the mini proteins were designed through computer modeling. Based on the results, five designed mini proteins were deemed to be viable stable mini proteins. They were then validated through experimental study. The smallest mini protein (model 5) was chosen for subsequent analysis as it was expressed in soluble form. No dehalogenase activity was detected, thus the specific binding interaction of between 1,3-dibromopropane with mini protein was investigated using isothermal titration calorimetry. Higher binding affinity between 1,3-dibromopropane and mini protein was obtained than the native. Thermal stability study with circular dichroism had proven that the mini protein possessed two times higher Tm value at 83.73 °C than the native at 43.97 °C. In conclusion, a stable mini protein was successfully designed and may be used as bioreceptors in the haloalkane sensor that is suitable for industrial application.

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Acknowledgements

We would like to thank Geran Universiti Putra Malaysia, Geran Putra Berkumpulan (IPB) (Project Number: GP-IPB/2013/9413500) for supporting our study.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

    Nurul Hazwani Daud & Siti Nurbaya Oslan

  2. Enzyme and Microbial Research Centre, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

    Nurul Hazwani Daud, Thean Chor Leow, Siti Nurbaya Oslan & Abu Bakar Salleh

  3. Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

    Thean Chor Leow

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  1. Nurul Hazwani Daud
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Correspondence to Abu Bakar Salleh.

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Daud, N.H., Leow, T.C., Oslan, S.N. et al. A Novel Mini Protein Design of Haloalkane Dehalogenase. Mol Biotechnol 61, 477–488 (2019). https://doi.org/10.1007/s12033-019-00169-3

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