High-yield production of FK228 and new derivatives in a Burkholderia chassis

Gong, Kai; Wang, Maoqin; Duan, Qiong; Li, Gang; Yong, Daojing; Ren, Cailing; Li, Yue; Zhang, Qi‐Jun; Wang, Zongjie; Sun, Tao; Zhang, Huanyun; Tu, Qiang; Wu, Changsheng; Fu, Jun; Li, Aiying; Song, Chaoyi; Zhang, Youming; Li, Rui-Juan

doi:10.1016/j.ymben.2022.12.002

Cited by 6 publications

(3 citation statements)

References 68 publications

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“…Four Burkholderia strains have been tested as heterologous hosts, with the soil isolate Burkholderia sp. FERM BP-3421 displaying a high perfomance. − The overall goal of our study is to enrich the genetic toolkit by exploring the regulation of Burkholderia metabolism while developing FERM BP-3421 as a synthetic biology chassis.…”

Section: Introductionmentioning

confidence: 99%

Understanding Autologous Spliceostatin Transcriptional Regulation to Derive Parts for Heterologous Expression in a Burkholderia Bacterial Host

2023

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Burkholderia β-Proteobacteria are emerging sources of natural products. We are interested in developing Burkholderia sp. FERM BP-3421 into a synthetic biology chassis to facilitate natural product discovery. FERM BP-3421 produces autologous spliceostatins on gram per liter scale. We reasoned that transcription factors and promoters that regulate spliceostatin biosynthesis would provide valuable parts for heterologous expression. Herein we demonstrate that fr9A encodes a pathway-specific transcriptional activator of spliceostatin biosynthesis. In-frame deletion of fr9A abolished spliceostatin production, which was restored by genetic complementation. Using transcriptomics and green fluorescent protein (GFP) reporter assays, we identified four fr9 promoters, three of which are activated by LuxR-type regulator Fr9A. We then constructed an Fr9A-regulated promoter system that was compared to benchmarks and effectively applied for GFP and capistruin lasso peptide expression in an optimized host background. Our findings enrich the genetic toolbox for optimizing heterologous expression and promoting the discovery and development of natural products from Burkholderia bacteria.

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Section: Introductionmentioning

confidence: 99%

Understanding Autologous Spliceostatin Transcriptional Regulation to Derive Parts for Heterologous Expression in a Burkholderia Bacterial Host

2023

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“…The Burkholderiales (class Betaproteobacteria ) are an emerging source of NPs and are multifaceted microorganisms diverse in their ecology and specialized metabolism. – A diverse range of NRPs or PKs have been mined from the Burkholderiales , such as the anticancer drug FK228 and its derivatives, , the pesticide lead pyrrolnitrin, , and the siderophores malleobactin, ornibactin, and caribactin. , Burkholderiales are also a potential chassis for expressing secondary metabolites and have attracted increasing attention . Recently, Schlegelella brevitalea DSM 7029, – Burkholderia thailandensis E264, , Burkholderia sp. FERM BP-3421, , Burkholderia gladioli pv.…”

Section: Introductionmentioning

confidence: 99%

“…25,26 Burkholderiales are also a potential chassis for expressing secondary metabolites and have attracted increasing attention. 27 Recently, Schlegelella brevitalea DSM 7029, 28−31 Burkholderia thailandensis E264, 22,32 Burkholderia sp. FERM BP-3421, 33,34 Burkholderia gladioli pv.…”

Section: ■ Introductionmentioning

confidence: 99%

Heterologous Biosynthesis of Complex Bacterial Natural Products in Burkholderia gladioli

Bai,

Chen,

Ren

et al. 2023

ACS Synth. Biol.

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Bacterial natural products (NPs) are an indispensable source of drugs and biopesticides. Heterologous expression is an essential method for discovering bacterial NPs and the efficient biosynthesis of valuable NPs, but the chassis for Gram-negative bacterial NPs remains inadequate. In this study, we built a Burkholderiales mutant Burkholderia gladioli Δgbn::attB by introducing an integrated site (attB) to inactivate the native gladiolin (gbn) biosynthetic gene cluster, which stabilizes large foreign gene clusters and reduces the native metabolite profile. The growth and successful heterologous production of high-value NPs such as phylogenetically close Burkholderiales-derived antitumor polyketides (PKs) rhizoxins, phylogenetically distant Gammaproteobacteria-derived anti-MRSA (methicillin-resistant Staphylococcus aureus) antibiotics WAP-8294As, and Deltaproteobacteria-derived antitumor PKs disorazols demonstrate that this strain is a potential chassis for Gram-negative bacterial NPs. We further improved the yields of WAP-8294As through promoter insertions and precursor pathway overexpression based on heterologous expression in this strain. This study provides a robust bacterial chassis for genome mining, efficient production, and molecular engineering of bacterial NPs.

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