A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution
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...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2020-06-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13068-020-01753-5 |
| _version_ | 1818229140970536960 |
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| author | Yilan Liu Jinjin Chen Anna N. Khusnutdinova Kevin Correia Patrick Diep Khorcheska A. Batyrova Kayla Nemr Robert Flick Peter Stogios Alexander F. Yakunin Radhakrishnan Mahadevan |
| author_facet | Yilan Liu Jinjin Chen Anna N. Khusnutdinova Kevin Correia Patrick Diep Khorcheska A. Batyrova Kayla Nemr Robert Flick Peter Stogios Alexander F. Yakunin Radhakrishnan Mahadevan |
| author_sort | Yilan Liu |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-12T10:13:53Z |
| format | Article |
| id | doaj.art-93d07098518a4a90be124e25301b96fc |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-12-12T10:13:53Z |
| publishDate | 2020-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-93d07098518a4a90be124e25301b96fc2022-12-22T00:27:43ZengBMCBiotechnology for Biofuels1754-68342020-06-0113111110.1186/s13068-020-01753-5A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolutionYilan Liu0Jinjin Chen1Anna N. Khusnutdinova2Kevin Correia3Patrick Diep4Khorcheska A. Batyrova5Kayla Nemr6Robert Flick7Peter Stogios8Alexander F. Yakunin9Radhakrishnan Mahadevan10Department of Chemical Engineering and Applied Chemistry, University of TorontoDepartment of Chemical Engineering and Applied Chemistry, University of TorontoDepartment of Chemical Engineering and Applied Chemistry, University of TorontoDepartment of Chemical Engineering and Applied Chemistry, University of TorontoDepartment of Chemical Engineering and Applied Chemistry, University of TorontoDepartment of Chemical Engineering and Applied Chemistry, University of TorontoDepartment of Chemical Engineering and Applied Chemistry, University of TorontoDepartment of Chemical Engineering and Applied Chemistry, University of TorontoDepartment of Chemical Engineering and Applied Chemistry, University of TorontoDepartment of Chemical Engineering and Applied Chemistry, University of TorontoDepartment of Chemical Engineering and Applied Chemistry, University of TorontoAbstract 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.http://link.springer.com/article/10.1186/s13068-020-01753-5AlkaneAldehyde decarbonylaseDirected evolutionDegradation tagProtease |
| spellingShingle | Yilan Liu Jinjin Chen Anna N. Khusnutdinova Kevin Correia Patrick Diep Khorcheska A. Batyrova Kayla Nemr Robert Flick Peter Stogios Alexander F. Yakunin Radhakrishnan Mahadevan A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution Biotechnology for Biofuels Alkane Aldehyde decarbonylase Directed evolution Degradation tag Protease |
| title | A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution |
| title_full | A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution |
| title_fullStr | A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution |
| title_full_unstemmed | A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution |
| title_short | A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution |
| title_sort | novel c terminal degron identified in bacterial aldehyde decarbonylases using directed evolution |
| topic | Alkane Aldehyde decarbonylase Directed evolution Degradation tag Protease |
| url | http://link.springer.com/article/10.1186/s13068-020-01753-5 |
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