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Luo Liu
Beijing University of Chemical Technology
Corresponding Author
ORCiD: https://orcid.org/0000-0002-3542-8213
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An active site normally locates inside of enzymes, substrates should go through the tunnel to access the active site. Tunnel engineering is a powerful strategy for refining the catalytic properties of enzymes. Here, P450 Bsβ HI (Q85H/ V170I) derived from hydroxylase P450 Bsβ from Bacillus subtilis was chosen as study model, which is reported as a potential decarbonylase. However, this enzyme showed low decarboxylase activity towards long-chain fatty acids. Here, a tunnel engineering campaign was performed for modulating the substrate preference and improving the decarbonylase activity of P450 Bsβ HI. The finally obtained BsβHI-F79A variant had a 15.2-fold improved conversion for palmitic acid; BsβHI-F173V variant had a 3.9-fold improved conversion for pentadecanoic acid. The study demonstrates how the substrate preference can be modulated by tunnel engineering strategy.
Keywords
Tunnel engineering, Substrate Preference, P450 decarbonylase, α-Alkene biosynthesis, Rational design
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CURRENT STATUS: Published
View the published article at Bioresources and Bioprocessing.
Version (no preprint)
Posted 23 Mar, 2021
Editorial decision: Accept
On 23 Mar, 2021
Submission checks complete
On 22 Mar, 2021
Editor invited
On 22 Mar, 2021
Editor assigned
On 02 Feb, 2021
First submitted
On 19 Jan, 2021
This version was not preprinted
Version (no preprint)
Posted 08 Mar, 2021
Review #2 received
Received 08 Mar, 2021
Reviewer #2 agreed
On 06 Mar, 2021
Editor assigned
On 01 Mar, 2021
Reviewers invited
Invitations sent on 01 Mar, 2021
Reviewer #1 agreed
On 01 Mar, 2021
Submission checks complete
On 01 Mar, 2021
Editor invited
On 01 Mar, 2021
This version was not preprinted
Version 2
Posted 27 Jan, 2021
No community comments so far
Review #3 received
Received 02 Feb, 2021
Editorial decision: Major revision
On 02 Feb, 2021
Review #2 received
Received 01 Feb, 2021
Reviewer #2 agreed
On 20 Jan, 2021
Reviewer #3 agreed
On 20 Jan, 2021
Reviewer #4 agreed
On 20 Jan, 2021
Review #4 received
Received 20 Jan, 2021
Reviewers invited
Invitations sent on 20 Jan, 2021
Reviewer #1 agreed
On 20 Jan, 2021
Review #1 received
Received 20 Jan, 2021
Editor assigned
On 19 Jan, 2021
Submission checks complete
On 19 Jan, 2021
Editor invited
On 19 Jan, 2021
No comments provided
Editorial decision: Major revision
On 08 Jan, 2021
Review #1 received
Received 06 Jan, 2021
Review #4 received
Received 27 Dec, 2020
Review #3 received
Received 20 Dec, 2020
Reviewer #4 agreed
On 20 Dec, 2020
Review #2 received
Received 20 Dec, 2020
Reviewer #3 agreed
On 19 Dec, 2020
Reviewer #2 agreed
On 19 Dec, 2020
Reviewers invited
Invitations sent on 18 Dec, 2020
Reviewer #1 agreed
On 18 Dec, 2020
Editor assigned
On 17 Dec, 2020
Submission checks complete
On 17 Dec, 2020
Editor invited
On 17 Dec, 2020
First submitted
On 14 Dec, 2020
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A new preprint design is coming!
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See the published version of this preprint at Bioresources and Bioprocessing.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Luo Liu
Beijing University of Chemical Technology
Corresponding Author
ORCiD: https://orcid.org/0000-0002-3542-8213
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at Bioresources and Bioprocessing.
Version (no preprint)
Posted 23 Mar, 2021
Editorial decision: Accept
On 23 Mar, 2021
Submission checks complete
On 22 Mar, 2021
Editor invited
On 22 Mar, 2021
Editor assigned
On 02 Feb, 2021
First submitted
On 19 Jan, 2021
This version was not preprinted
Version (no preprint)
Posted 08 Mar, 2021
Review #2 received
Received 08 Mar, 2021
Reviewer #2 agreed
On 06 Mar, 2021
Editor assigned
On 01 Mar, 2021
Reviewers invited
Invitations sent on 01 Mar, 2021
Reviewer #1 agreed
On 01 Mar, 2021
Submission checks complete
On 01 Mar, 2021
Editor invited
On 01 Mar, 2021
This version was not preprinted
Version 2
Posted 27 Jan, 2021
No community comments so far
Review #3 received
Received 02 Feb, 2021
Editorial decision: Major revision
On 02 Feb, 2021
Review #2 received
Received 01 Feb, 2021
Reviewer #2 agreed
On 20 Jan, 2021
Reviewer #3 agreed
On 20 Jan, 2021
Reviewer #4 agreed
On 20 Jan, 2021
Review #4 received
Received 20 Jan, 2021
Reviewers invited
Invitations sent on 20 Jan, 2021
Reviewer #1 agreed
On 20 Jan, 2021
Review #1 received
Received 20 Jan, 2021
Editor assigned
On 19 Jan, 2021
Submission checks complete
On 19 Jan, 2021
Editor invited
On 19 Jan, 2021
No comments provided
Editorial decision: Major revision
On 08 Jan, 2021
Review #1 received
Received 06 Jan, 2021
Review #4 received
Received 27 Dec, 2020
Review #3 received
Received 20 Dec, 2020
Reviewer #4 agreed
On 20 Dec, 2020
Review #2 received
Received 20 Dec, 2020
Reviewer #3 agreed
On 19 Dec, 2020
Reviewer #2 agreed
On 19 Dec, 2020
Reviewers invited
Invitations sent on 18 Dec, 2020
Reviewer #1 agreed
On 18 Dec, 2020
Editor assigned
On 17 Dec, 2020
Submission checks complete
On 17 Dec, 2020
Editor invited
On 17 Dec, 2020
First submitted
On 14 Dec, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Biotechnology and Bioengineering
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Bioresources and Bioprocessing.
An active site normally locates inside of enzymes, substrates should go through the tunnel to access the active site. Tunnel engineering is a powerful strategy for refining the catalytic properties of enzymes. Here, P450 Bsβ HI (Q85H/ V170I) derived from hydroxylase P450 Bsβ from Bacillus subtilis was chosen as study model, which is reported as a potential decarbonylase. However, this enzyme showed low decarboxylase activity towards long-chain fatty acids. Here, a tunnel engineering campaign was performed for modulating the substrate preference and improving the decarbonylase activity of P450 Bsβ HI. The finally obtained BsβHI-F79A variant had a 15.2-fold improved conversion for palmitic acid; BsβHI-F173V variant had a 3.9-fold improved conversion for pentadecanoic acid. The study demonstrates how the substrate preference can be modulated by tunnel engineering strategy.
Figure 1
Figure 2
Figure 3
Figure 4
This is a list of supplementary files associated with this preprint. Click to download.
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