A Novel Mini Protein Design of Haloalkane Dehalogenase

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. Therefo...

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Bibliographic Details
Published in:Molecular biotechnology 2019-07, Vol.61 (7), p.477-488
Main Authors: Daud, Nurul Hazwani, Leow, Thean Chor, Oslan, Siti Nurbaya, Salleh, Abu Bakar
Format: Article
Language:English
Subjects:
Binding
Biochemistry
Biological Techniques
Biotechnology
Calorimetry
Cell Biology
Chemistry
Chemistry and Materials Science
Circular dichroism
Contaminants
Dichroism
Haloalkane dehalogenase
Human Genetics
Industrial applications
Original Paper
Protein Science
Proteins
Thermal stability
Titration
Titration calorimetry
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Summary: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 T m 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.
ISSN:1073-6085
1559-0305
DOI:10.1007/s12033-019-00169-3