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Biosensor-based enzyme engineering approach applied to psicose biosynthesis.

Jeremy Armetta¹, Rose Berthome¹, Antonin Cros¹

¹iSSB, UMR8030 Génomique Métabolique, Genoscope, Institut François Jacob, CEA, CNRS, Univ Evry, Université Paris-Saclay, Genopole Campus 1, Bât. 6, 5 rue Henri Desbruères, 91030 Evry, France.

Synthetic Biology (Oxford, England)

|September 30, 2020

Summary

This study introduces a novel biosensor framework for efficiently screening enzymes, accelerating the bioproduction of D-psicose (D-allulose), a rare sugar with potential health benefits.

Keywords:

Universal Biosensing Chassis enzyme engineering psicose rare sugars transcription factor-based biosensor

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Area of Science:

Synthetic biology
Biotechnology
Enzyme engineering

Background:

Bioproduction offers cost-effective and eco-friendly alternatives for chemical manufacturing.
D-psicose (D-allulose) is a rare sugar with potential therapeutic applications for obesity and type 2 diabetes.
Efficient enzyme screening is crucial for optimizing bioproduction yields.

Purpose of the Study:

To develop and validate a biosensor-based screening approach for enzyme selection in D-psicose bioproduction.
To engineer and characterize novel D-psicose-responsive biosensors.
To demonstrate the framework's utility in identifying improved D-psicose 3-epimerase variants.

Main Methods:

Construction and characterization of seven D-psicose-responsive biosensors using transcription factors and promoters.
Development of the Universal Biosensor Chassis for modular biosensor engineering.
Error-prone PCR amplification and high-throughput screening of D-psicose 3-epimerase mutant libraries using the selected biosensor.

Main Results:

Seven novel D-psicose biosensors were successfully constructed and characterized.
A highly responsive and linear biosensor was selected for enzyme screening.
The biosensor framework enabled the identification of gain-of-function D-psicose 3-epimerase mutants.

Conclusions:

The developed biosensor-based screening framework significantly enhances enzyme discovery for D-psicose bioproduction.
This approach facilitates the optimization of biocatalysts for cost-effective and sustainable chemical manufacturing.
The Universal Biosensor Chassis provides a versatile platform for future biosensor engineering applications.