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Updated: Feb 24, 2026
A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates
Published on: August 21, 2018
Modelling of microbial polyhydroxyalkanoate surface binding protein PhaP for rational mutagenesis
Hongyu Zhao1, Zhenyu Yao1, Xiangbin Chen1
1Center for Synthetic and Systems Biology, School of Life Sciences, Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing, China.
Phasins are microbial proteins binding to polyhydroxyalkanoate (PHA) granules. Mutagenesis of homology models enhanced their stability and surfactant properties, showing potential for biotechnology applications.
Area of Science:
- Biochemistry
- Protein Engineering
Background:
- Phasins are amphiphilic proteins that bind to microbial polyhydroxyalkanoate (PHA) granules.
- These proteins have diverse applications in biotechnology and medicine.
- The crystal structure of PhaPAh is the only one solved to date.
Purpose of the Study:
- To establish homology models for PhaPAz and PhaPTD.
- To enhance the stability and surfactant properties of these phasins through rational mutagenesis.
- To generalize the homology-modelling approach for studying other PhaP members.
Main Methods:
- Homology modelling based on the crystal structure of PhaPAh.
- Rational mutagenesis of PhaPAz and PhaPTD.
- Evaluation of emulsification properties and thermostability of wild-type and mutant proteins.
Main Results:
- Homology models for PhaPAz and PhaPTD were successfully established.
- Mutants PhaPAz Q38L, PhaPAz Q78L, PhaPTD Q38M, and PhaPTD Q72M exhibited improved emulsification.
- Mutants showed increased thermostability, with 6-10°C higher melting temperatures compared to wild-type proteins.
Conclusions:
- The homology-modelling approach, based on PhaPAh structure, is effective for studying other PhaP proteins.
- Engineered phasins demonstrate enhanced stability and surfactant capabilities.
- This study facilitates further research and application development for diverse phasin proteins.
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