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Cloning and expression of Burkholderia polyyne biosynthetic gene clusters in Paraburkholderia hosts provides a strategy for biopesticide development
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Petrova, Yoana D., Zhao, Jinlian, Webster, Gordon, Mullins, Alex J., Williams, Katherine, Alswat, Amal S., Challis, Gregory L., Bailey, Andy M. and Mahenthiralingam, Eshwar (2022) Cloning and expression of Burkholderia polyyne biosynthetic gene clusters in Paraburkholderia hosts provides a strategy for biopesticide development. Microbial Biotechnology, 15 (10). pp. 2547-2561. doi:10.1111/1751-7915.14106 ISSN 1751-7907.
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WRAP-cloning-expression-Burkholderia-polyyne-biosynthetic-gene-clusters-Paraburkholderia-hosts-provides-strategy-biopesticide-development-Challis-2022.pdf - Published Version - Requires a PDF viewer. Available under License Creative Commons Attribution 4.0. Download (4065Kb) | Preview |
Official URL: https://doi.org/10.1111/1751-7915.14106
Abstract
Burkholderia have potential as biocontrol agents because they encode diverse biosynthetic gene clusters (BGCs) for a range of antimicrobial metabolites. Given the opportunistic pathogenicity associated with Burkholderia species, heterologous BGC expression within non‐pathogenic hosts is a strategy to construct safe biocontrol strains. We constructed a yeast‐adapted Burkholderia‐Escherichia shuttle vector (pMLBAD_yeast) with a yeast replication origin 2 μ and URA3 selection marker and optimised it for cloning BGCs using the in vivo recombination ability of Saccharomyces cerevisiae. Two Burkholderia polyyne BGCs, cepacin (13 kb) and caryoynencin (11 kb), were PCR‐amplified as three overlapping fragments, cloned downstream of the pBAD arabinose promoter in pMLBAD_yeast and mobilised into Burkholderia and Paraburkholderia heterologous hosts. Paraburkholderia phytofirmans carrying the heterologous polyyne constructs displayed in vitro bioactivity against a variety of fungal and bacterial plant pathogens similar to the native polyyne producers. Thirteen Paraburkholderia strains with preferential growth at 30°C compared with 37°C were also identified, and four of these were amenable to genetic manipulation and heterologous expression of the caryoynencin construct. The cloning and successful heterologous expression of Burkholderia biosynthetic gene clusters within Paraburkholderia with restricted growth at 37°C opens avenues for engineering non‐pathogenic biocontrol strains.
| Item Type: | Journal Article | ||||||||||||
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| Subjects: | Q Science > QR Microbiology T Technology > TP Chemical technology |
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| Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry | ||||||||||||
| SWORD Depositor: | Library Publications Router | ||||||||||||
| Library of Congress Subject Headings (LCSH): | Ralstonia solanacearum, Natural pesticides, Microbial biotechnology | ||||||||||||
| Journal or Publication Title: | Microbial Biotechnology | ||||||||||||
| Publisher: | Wiley-Blackwell Publishing Ltd. | ||||||||||||
| ISSN: | 1751-7907 | ||||||||||||
| Official Date: | October 2022 | ||||||||||||
| Dates: |
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| Volume: | 15 | ||||||||||||
| Number: | 10 | ||||||||||||
| Page Range: | pp. 2547-2561 | ||||||||||||
| DOI: | 10.1111/1751-7915.14106 | ||||||||||||
| Status: | Peer Reviewed | ||||||||||||
| Publication Status: | Published | ||||||||||||
| Access rights to Published version: | Open Access (Creative Commons) | ||||||||||||
| Date of first compliant deposit: | 12 August 2022 | ||||||||||||
| Date of first compliant Open Access: | 24 August 2022 | ||||||||||||
| RIOXX Funder/Project Grant: |
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