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A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution
Yilan Liu1, Jinjin Chen1, Anna N Khusnutdinova1
1Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, ON M5S 3E5 Canada.
Biotechnology for Biofuels
|July 3, 2020
Summary
Scientists identified a degradation tag (degron) that destabilizes aldehyde decarbonylases (ADs). Modifying or removing this tag enhances AD stability, leading to increased alkane production from renewable sources.
Area of Science:
- Biochemistry
- Molecular Biology
- Synthetic Biology
Background:
- Aldehyde decarbonylases (ADs) convert acyl aldehydes to alkanes, offering a renewable feedstock conversion pathway.
- The inherent instability of ADs limits their practical application in alkane production.
Purpose of the Study:
- Investigate the degradation mechanism of ADs.
- Engineer ADs for enhanced stability and improved alkane yields.
Main Methods:
- Error-prone PCR-based directed evolution to identify degradation tags.
- Bioinformatic analysis to identify conserved degron motifs.
- In vivo studies in E. coli to assess AD stability and alkane production.
Main Results:
- Discovered a C-terminal degradation tag (degron) in aldehyde decarbonylases from Prochlorococcus marinus.
- Identified a conserved C-terminal motif (RMSAYGLAAA) as the AD degron (ADcon).
- Demonstrated that ATP-dependent proteases ClpAP and Lon mediate AD degradation, limiting alkane production.
- Deletion or modification of the degron significantly increased in vivo alkane production.
Conclusions:
- Revealed a novel degron in bacterial ADs responsible for their instability.
- Eliminating or modifying the degron stabilizes ADs, enabling higher alkane titers.
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