A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution
Posted on 2020-06-30 - 04:29
Abstract Background Aldehyde decarbonylases (ADs), which convert acyl aldehydes into alkanes, supply promising solution for producing alkanes from renewable feedstock. However the instability of ADs impedes their further application. Therefore, the current study aimed to investigate the degradation mechanism of ADs and engineer it towards high stability. Results Here, we describe the discovery of a degradation tag (degron) in the AD from marine cyanobacterium Prochlorococcus marinus using error-prone PCR-based directed evolution system. Bioinformatic analysis revealed that this C-terminal degron is common in bacterial ADs and identified a conserved C-terminal motif, RMSAYGLAAA, representing the AD degron (ADcon). Furthermore, we demonstrated that the ATP-dependent proteases ClpAP and Lon are involved in the degradation of AD-tagged proteins in E. coli, thereby limiting alkane production. Deletion or modification of the degron motif increased alkane production in vivo. Conclusion This work revealed the presence of a novel degron in bacterial ADs responsible for its instability. The in vivo experiments proved eliminating or modifying the degron could stabilize AD, thereby producing higher titers of alkanes.
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Liu, Yilan; Chen, Jinjin; Khusnutdinova, Anna N.; Correia, Kevin; Diep, Patrick; Batyrova, Khorcheska A.; et al. (2020). A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution. figshare. Collection. https://doi.org/10.6084/m9.figshare.c.5044478.v1
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AUTHORS (11)
YL
Yilan Liu
JC
Jinjin Chen
AK
Anna N. Khusnutdinova
KC
Kevin Correia
PD
Patrick Diep
KB
Khorcheska A. Batyrova
KN
Kayla Nemr
RF
Robert Flick
PS
Peter Stogios
AY
Alexander F. Yakunin
RM
Radhakrishnan Mahadevan