Cloning and expression of Burkholderia polyyne biosynthetic gene clusters in Paraburkholderia hosts provides a strategy for biopesticide development.

Autor: Petrova YD; School of Biosciences, Cardiff University, Cardiff, UK., Zhao J; Department of Chemistry, University of Warwick, Coventry, UK., Webster G; School of Biosciences, Cardiff University, Cardiff, UK., Mullins AJ; School of Biosciences, Cardiff University, Cardiff, UK., Williams K; School of Biological Sciences, University of Bristol, Bristol, UK., Alswat AS; School of Biosciences, Cardiff University, Cardiff, UK., Challis GL; Department of Chemistry, University of Warwick, Coventry, UK.; Warwick Integrative Synthetic Biology Centre, University of Warwick, Coventry, UK.; Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia.; ARC Centre of Excellence for Innovations in Peptide and Protein Science, Monash University, Clayton, Victoria, Australia., Bailey AM; School of Biological Sciences, University of Bristol, Bristol, UK., Mahenthiralingam E; School of Biosciences, Cardiff University, Cardiff, UK.
Jazyk: angličtina
Zdroj: Microbial biotechnology [Microb Biotechnol] 2022 Oct; Vol. 15 (10), pp. 2547-2561. Date of Electronic Publication: 2022 Jul 13.
DOI: 10.1111/1751-7915.14106
Abstrakt: 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.
(© 2022 The Authors. Microbial Biotechnology published by Society for Applied Microbiology and John Wiley & Sons Ltd.)
Databáze: MEDLINE
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