Targeted activation of silent natural product biosynthesis pathways by reporter-guided mutant selection

Guo, Fang; Xiang, Sihai; Li, Liyuan; Wang, Bin; Rajasärkkä, Johanna; Gröndahl-Yli-Hannuksela, Kirsi; Ai, Guomin; Metsä‐Ketelä, Mikko; Yang, Keqian

doi:10.1016/j.ymben.2014.12.006

Cited by 71 publications

(70 citation statements)

References 52 publications

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“…In a recent report Guo et al constructed a reporter system linking the expression of genes for gaudimycin biosynthesis to a reporter system in Streptomyces 10 . However, authors have reported high percentage of false-positives which, while expressing a reporter, did not actually produced a target metabolite.…”

Section: Discussionmentioning

confidence: 99%

“…Such biosensor would represent a clear advantage over the earlier described reporter system that detects expression of the secondary metabolite biosynthesis genes, as described by Guo et al for gaudimycin biosynthesis. 10 Indeed, our design would allow direct monitoring of the cluster-specific product, not just expression of its biosynthetic genes. Here, we present a proof-of-principle for such a biosensor, and argue that similar design can greatly assist in the genome-based biosprospecting for novel bioactive compounds in Streptomyces and other actinomycete bacteria.…”

mentioning

confidence: 99%

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Development of a Biosensor Concept to Detect the Production of Cluster-Specific Secondary Metabolites

et al. 2017

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Genome mining of actinomycete bacteria aims at the discovery of novel bioactive secondary metabolites that can be developed into drugs. A new repressorbased biosensor to detect activated secondary metabolite biosynthesis gene clusters in Streptomyces was developed. Biosynthetic gene clusters for undecylprodigiosin and coelimycin in the genome of Streptomyces lividans TK24, which encoded TetR-like repressors and appeared to be almost "silent" based on the RNA-seq data, were chosen for the proof-of-principle studies. The bpsA reporter gene for indigoidine synthetase was placed under control of the promotor/operator regions presumed to be controlled by the cluster-associated TetR-like repressors. While the biosensor for undecylprodigiosin turned out to be nonfunctional, the coelimycin biosensor was shown to perform as expected, turning on biosynthesis of indigoidine in response to the concomitant production of coelimycin. The developed reporter system concept can be applied to those cryptic gene clusters that encode metabolite-sensing repressors to speed up discovery of novel bioactive compounds in Streptomyces.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Development of a Biosensor Concept to Detect the Production of Cluster-Specific Secondary Metabolites

et al. 2017

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“…However, it should be noted that, despite all these resources available, a certain amount of iterations of the design-build-test-learn cycle are still necessary as demonstrated for other microbial metabolic engineering cases, especially taking into account the complex regulation between primary and secondary metabolism in streptomycetes. Implementation of molecular cloning methods in an automated, high-throughput setting, such as iBioFab, [80] biosensors for detecting expression of secondary metabolite biosynthesis genes, [81] or biosensors for the production of a desired secondary metabolite [82] might further reduce time and efforts needed to optimize the production of secondary metabolites using streptomycetes. It is expected that the tools and strategies of systems metabolic engineering of streptomycetes will advance rapidly to harness full biotechnological potentials of this important class of bacteria.…”

Section: Discussionmentioning

confidence: 99%

Toward Systems Metabolic Engineering of Streptomycetes for Secondary Metabolites Production

Robertsen

Weber

Kim

et al. 2017

Biotechnology Journal

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Streptomycetes are known for their inherent ability to produce pharmaceutically relevant secondary metabolites. Discovery of medically useful, yet novel compounds has become a great challenge due to frequent rediscovery of known compounds and a consequent decline in the number of relevant clinical trials in the last decades. A paradigm shift took place when the first whole genome sequences of streptomycetes became available, from which silent or "cryptic" biosynthetic gene clusters (BGCs) were discovered. Cryptic BGCs reveal a so far untapped potential of the microorganisms for the production of novel compounds, which has spurred new efforts in understanding the complex regulation between primary and secondary metabolism. This new trend has been accompanied with development of new computational resources (genome and compound mining tools), generation of various high-quality omics data, establishment of molecular tools, and other strain engineering strategies. They all come together to enable systems metabolic engineering of streptomycetes, allowing more systematic and efficient strain development. In this review, the authors present recent progresses within systems metabolic engineering of streptomycetes for uncovering their hidden potential to produce novel compounds and for the improved production of secondary metabolites.

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“…Moreover, two novel chattamycins were discovered when overexpressing an atypical response regulator (ARR) family regulator gene, chaI [14]; and overexpression of a LuxR family activator gene nam1 led to production of three new naphthalenic octaketide ansamycins [15]. For the jadomycin and gaudimycin biosynthetic pathways, however, no significant effects were observed when ARR family activator genes were overexpressed [16 • ]. This approach is proved to be pathway-specific, thus the efficacy varies case by case.…”

Section: Activation Of Silent Bgcs In Native Hostsmentioning

confidence: 99%

Breaking the silence: new strategies for discovering novel natural products

Ren

Wang

Zhao

2017

Current Opinion in Biotechnology

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103

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Natural products have been a prolific source of antibacterial and anticancer drugs for decades. One of the major challenges in natural product discovery is that the vast majority of natural product biosynthetic gene clusters (BGCs) have not been characterized, partially due to the fact that they are either transcriptionally silent or expressed at very low levels under standard laboratory conditions. Here we describe the strategies developed in recent years (mostly between 2014–2016) for activating silent BGCs. These strategies can be broadly divided into two categories: approaches in native hosts and approaches in heterologous hosts. In addition, we briefly discuss recent advances in developing new computational tools for identification and characterization of BGCs and high-throughput methods for detection of natural products.

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Targeted activation of silent natural product biosynthesis pathways by reporter-guided mutant selection

Cited by 71 publications

References 52 publications

Development of a Biosensor Concept to Detect the Production of Cluster-Specific Secondary Metabolites

Development of a Biosensor Concept to Detect the Production of Cluster-Specific Secondary Metabolites

Toward Systems Metabolic Engineering of Streptomycetes for Secondary Metabolites Production

Breaking the silence: new strategies for discovering novel natural products

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