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Xiaoying Liu
Anhui Medical University
Corresponding Author
Qi Chen
Anhui Medical University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0532-7390
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This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Microbial Cell Factories.
Backgrounds: Activation of silent biosynthetic gene clusters (BGCs) in marine-derived actinomycete strains is a feasible strategy to discover bioactive natural products. Actinoalloteichus sp. AHMU CJ021, isolated from the seashore, was shown to contain an intact but silent caerulomycin A (CRM A) BGC- cam in its genome. Thus, a genome mining work was preformed to activate the strain’s production of CRM A, an immunosuppressive drug lead with diverse bioactivities.
Results: To well activate the expression of cam , ribosome engineering was adopted to treat the wild type Actinoalloteichus sp. AHMU CJ021. The initial mutant strain XC-11G with gentamycin resistance and CRM A production titer of 42.51 ± 4.22 mg/L was selected from all generated mutant strains by gene expression comparison of the essential biosynthetic gene-camE. The titer of CRM A production was then improved by two strain breeding methods via UV mutagenesis and cofactor engineering-directed increase of intracellular riboflavin, which finally generated the optimal mutant strain XC-11GUR with a CRM A production titer of 113.91 ± 7.58 mg/L. Subsequently, this titer of strain XC-11GUR was improved to 618.61 ± 16.29 mg/L through medium optimization together with further adjustment derived from response surface methodology. In terms of this 14.6 folds increase in the titer of CRM A compared to the initial value, strain XC-GUR could be a well alternative strain for CRM A development.
Conclusions: Our results have constructed an ideal CRM A producer. More importantly, our efforts also have demonstrated the effectiveness of abovementioned combinatorial strategies, which is applicable to the genome mining of bioactive natural products from abundant actinomycetes strains.
Keywords
Caerulomycin A, Genome mining, Combinatorial strategies, Ribosome engineering, Strain improvement, Marine-derived Actinoalloteichus
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CURRENT STATUS: Published
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Posted 30 Jul, 2020
Published
On 06 Aug, 2020
No community comments so far
Editorial decision: Accept
On 28 Jul, 2020
Reviewer #1 agreed
On 26 Jul, 2020
Review #1 received
Received 26 Jul, 2020
Editor assigned
On 24 Jul, 2020
Reviewers invited
Invitations sent on 24 Jul, 2020
Submission checks complete
On 23 Jul, 2020
Editor invited
On 23 Jul, 2020
No comments provided
Editorial decision: Major Revision
On 20 Jul, 2020
Review #2 received
Received 19 Jul, 2020
Review #3 received
Received 17 Jul, 2020
Reviewer #3 agreed
On 12 Jul, 2020
Reviewer #2 agreed
On 11 Jul, 2020
Review #1 received
Received 08 Jul, 2020
Editor assigned
On 07 Jul, 2020
Reviewers invited
Invitations sent on 07 Jul, 2020
Reviewer #1 agreed
On 07 Jul, 2020
Submission checks complete
On 06 Jul, 2020
Editor invited
On 06 Jul, 2020
No comments provided
Editorial decision: Major Revision
On 29 May, 2020
Review #2 received
Received 22 May, 2020
Review #3 received
Received 20 May, 2020
Review #1 received
Received 18 May, 2020
Reviewer #4 agreed
On 12 May, 2020
Reviewer #3 agreed
On 12 May, 2020
Reviewer #2 agreed
On 11 May, 2020
Reviewer #1 agreed
On 08 May, 2020
Reviewers invited
Invitations sent on 07 May, 2020
Editor assigned
On 04 May, 2020
First submitted
On 03 May, 2020
Submission checks complete
On 03 May, 2020
Editor invited
On 03 May, 2020
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Subject Areas
Applied & Industrial Microbiology
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This preprint is under consideration at Microbial Cell Factories. A preprint is 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
Xiaoying Liu
Anhui Medical University
Corresponding Author
Qi Chen
Anhui Medical University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0532-7390
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
Version 3
Posted 30 Jul, 2020
Published
On 06 Aug, 2020
No community comments so far
Editorial decision: Accept
On 28 Jul, 2020
Reviewer #1 agreed
On 26 Jul, 2020
Review #1 received
Received 26 Jul, 2020
Editor assigned
On 24 Jul, 2020
Reviewers invited
Invitations sent on 24 Jul, 2020
Submission checks complete
On 23 Jul, 2020
Editor invited
On 23 Jul, 2020
No comments provided
Editorial decision: Major Revision
On 20 Jul, 2020
Review #2 received
Received 19 Jul, 2020
Review #3 received
Received 17 Jul, 2020
Reviewer #3 agreed
On 12 Jul, 2020
Reviewer #2 agreed
On 11 Jul, 2020
Review #1 received
Received 08 Jul, 2020
Editor assigned
On 07 Jul, 2020
Reviewers invited
Invitations sent on 07 Jul, 2020
Reviewer #1 agreed
On 07 Jul, 2020
Submission checks complete
On 06 Jul, 2020
Editor invited
On 06 Jul, 2020
No comments provided
Editorial decision: Major Revision
On 29 May, 2020
Review #2 received
Received 22 May, 2020
Review #3 received
Received 20 May, 2020
Review #1 received
Received 18 May, 2020
Reviewer #4 agreed
On 12 May, 2020
Reviewer #3 agreed
On 12 May, 2020
Reviewer #2 agreed
On 11 May, 2020
Reviewer #1 agreed
On 08 May, 2020
Reviewers invited
Invitations sent on 07 May, 2020
Editor assigned
On 04 May, 2020
First submitted
On 03 May, 2020
Submission checks complete
On 03 May, 2020
Editor invited
On 03 May, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Applied & Industrial Microbiology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Microbial Cell Factories.
Backgrounds: Activation of silent biosynthetic gene clusters (BGCs) in marine-derived actinomycete strains is a feasible strategy to discover bioactive natural products. Actinoalloteichus sp. AHMU CJ021, isolated from the seashore, was shown to contain an intact but silent caerulomycin A (CRM A) BGC- cam in its genome. Thus, a genome mining work was preformed to activate the strain’s production of CRM A, an immunosuppressive drug lead with diverse bioactivities.
Results: To well activate the expression of cam , ribosome engineering was adopted to treat the wild type Actinoalloteichus sp. AHMU CJ021. The initial mutant strain XC-11G with gentamycin resistance and CRM A production titer of 42.51 ± 4.22 mg/L was selected from all generated mutant strains by gene expression comparison of the essential biosynthetic gene-camE. The titer of CRM A production was then improved by two strain breeding methods via UV mutagenesis and cofactor engineering-directed increase of intracellular riboflavin, which finally generated the optimal mutant strain XC-11GUR with a CRM A production titer of 113.91 ± 7.58 mg/L. Subsequently, this titer of strain XC-11GUR was improved to 618.61 ± 16.29 mg/L through medium optimization together with further adjustment derived from response surface methodology. In terms of this 14.6 folds increase in the titer of CRM A compared to the initial value, strain XC-GUR could be a well alternative strain for CRM A development.
Conclusions: Our results have constructed an ideal CRM A producer. More importantly, our efforts also have demonstrated the effectiveness of abovementioned combinatorial strategies, which is applicable to the genome mining of bioactive natural products from abundant actinomycetes strains.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
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