Related Experiment Video
Updated: Oct 5, 2025
09:16
FIBS-enabled Noninvasive Metabolic Profiling
Published on: February 3, 2014
10.0K
Discovery of Thermostable, Fluorescently Responsive Glucose Biosensors by Structure-Assisted Function Extrapolation
Malin J Allert1, Homme W Hellinga1
1Department of Biochemistry, Duke University Medical Center, Box 3711, Durham, North Carolina 27710, United States.
Biochemistry
|January 27, 2022
Summary
Researchers discovered thermostable glucose-binding proteins for improved biosensors. A structure-based filter accurately identifies glucose-binding paralogs, enabling the development of next-generation fluorescent glucose sensors.
Area of Science:
- Biochemistry
- Structural Biology
- Genomics
Background:
- The periplasmic-binding protein (PBP) superfamily is ubiquitous in prokaryotes and prone to gene duplication, leading to numerous paralogs.
- Engineered glucose-binding PBPs from Escherichia coli serve as reagentless fluorescent biosensors.
- Developing robust biosensors for real-world applications requires identifying thermostable homologues with retained function.
Purpose of the Study:
- To discover thermostable periplasmic-binding protein (PBP) homologues that function as fluorescent glucose biosensors.
- To develop and validate a structure-based method for accurately identifying glucose-binding PBPs among paralogs.
- To identify suitable candidates for next-generation robust fluorescent glucose sensors.
Main Methods:
- A structure-based filtering approach was employed, focusing on residues critical for glucose binding in an archetype structure.
- The filter was applied to fully sequenced bacterial genomes to assess its effectiveness in reducing paralog numbers.
- Engineered proteins of eight selected homologues were expressed and tested for glucose-mediated fluorescence response.
Main Results:
- The structure-based filter effectively reduced high paralog numbers to single genome hits, indicating accurate functional separation.
- Glucose binding was accurately predicted down to 31% sequence identity among tested homologues.
- Eight engineered homologues were successfully expressed and characterized for their glucose-mediated fluorescence response.
Conclusions:
- A structure-based sequence filtering method is highly effective for identifying specific glucose-binding periplasmic-binding protein (PBP) homologues.
- Thermostable PBP homologues with retained glucose-mediated fluorescence responses have been identified as candidates for advanced biosensors.
- This work facilitates the development of robust, next-generation fluorescent glucose sensors for diverse applications.
