Recombinational cloning of the antibiotic biosynthetic gene clusters in linear plasmid SCP1 of<i>Streptomyces coelicolor</i>A3(2)

Zhang, Ran; Xia, Haiyang; Xu, Qingyu; Dang, Fujun; Qin, Zhongjun

doi:10.1111/1574-6968.12183

Cited by 6 publications

(6 citation statements)

References 52 publications

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“… 81 , 82 Avermectin production was detected by using a new recombination cloning method to clone the 81-kb avermectin biosynthetic gene cluster into the linear plasmid of the model organism S. coelicolor . 83 This confirms that the avermectin metabolite pathway is available in the new host S. coelicolor .…”

Section: Synthetic Biology Methodssupporting

confidence: 60%

Interrogation of Streptomyces avermitilis for efficient production of avermectins

Chen

Liu

et al. 2016

Synthetic and Systems Biotechnology

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The 2015 Nobel Prize in Physiology or Medicine has been awarded to avermectins and artemisinin, respectively. Avermectins produced by Streptomyces avermitilis are excellent anthelmintic and potential antibiotic agents. Because wild-type strains only produce low levels of avermectins, much research effort has focused on improvements in avermectin production to meet the ever increasing demand for such compounds. This review describes the strategies that have been widely employed and the future prospects of synthetic biology applications in avermectin yield improvement. With the help of genome sequencing of S. avermitilis and an understanding of the avermectin biosynthetic/regulatory pathways, synthetic and systems biotechnology approaches have been applied for precision engineering. We focus on the design and synthesis of biological chassis, parts, devices, and modules from diverse microbes to reconstruct and optimize their dynamic processes, as well as predict favorable effective overproduction of avermectins by a 4Ms strategy (Mine, Model, Manipulation, and Measurement).

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Section: Synthetic Biology Methodssupporting

confidence: 60%

Interrogation of Streptomyces avermitilis for efficient production of avermectins

Chen

Liu

et al. 2016

Synthetic and Systems Biotechnology

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“…40 Lastly, but most importantly, probably due to such a complicated and delicate postmodification, previous attempts on heterologous production of spinosyn in Streptomyces were ended in failure. 41 In this work, a set of strategies involved in BAC library construction, efficient conjugation and integration of a large plasmid into a Streptomyces host, identification of rate-limiting steps by omics analysis, and rational synthetic module optimization for production of targeted products in a heterologous host have been developed. This resulting approach is likewise expected to become adaptable to highly efficient heterologous production of valuable NPs with complex structures.…”

mentioning

confidence: 99%

“…In the 80-kb spinosyn biosynthetic gene cluster, 14 of 19 genes are responsible for spinosyn postmodification, which include complicated intramolecular cycloaddition , and cross-bridging, as well as multiple steps of glycosylation − and methylation . Lastly, but most importantly, probably due to such a complicated and delicate postmodification, previous attempts on heterologous production of spinosyn in Streptomyces were ended in failure …”

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confidence: 99%

Heterologous Biosynthesis of Spinosad: An Omics-Guided Large Polyketide Synthase Gene Cluster Reconstitution in Streptomyces

et al. 2017

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With the advent of the genomics era, heterologous gene expression has been used extensively as a means of accessing natural products (NPs) from environmental DNA samples. However, the heterologous production of NPs often has very low efficiency or is unable to produce targeted NPs. Moreover, due to the complicated transcriptional and metabolic regulation of NP biosynthesis in native producers, especially in the cases of genome mining, it is also difficult to rationally and systematically engineer synthetic pathways to improved NPs biosynthetic efficiency. In this study, various strategies ranging from heterologous production of a NP to subsequent application of omics-guided synthetic modules optimization for efficient biosynthesis of NPs with complex structure have been developed. Heterologous production of spinosyn in Streptomyces spp. has been demonstrated as an example of the application of these approaches. Combined with the targeted omics approach, several rate-limiting steps of spinosyn heterologous production in Streptomyces spp. have been revealed. Subsequent engineering work overcame three of selected rate-limiting steps, and the production of spinosad was increased step by step and finally reached 1460 μg/L, which is about 1000-fold higher than the original strain S. albus J1074 (C4I6-M). These results indicated that the omics platform developed in this work was a powerful tool for guiding the rational refactoring of heterologous biosynthetic pathway in Streptomyces host. Additionally, this work lays the foundation for further studies aimed at the more efficient production of spinosyn in a heterologous host. And the strategy developed in this study is expected to become readily adaptable to highly efficient heterologous production of other NPs with complex structure.

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“…Streptomyces is the most important source of NPs and produces kinds of cellular intermediates of NPs with complex structures (Komatsu et al ., 2013 ; Tan and Liu, 2017 ). Consequently, heterologous production of NPs is preferably performed in Streptomyces sp., especially in the strains with fully sequenced genome such as Streptomyces coelicolor (Jones et al ., 2013 ; Zhang et al ., 2013 ).…”

Section: Introductionmentioning

confidence: 99%

Multiplying the heterologous production of spinosad through tandem amplification of its biosynthetic gene cluster in Streptomyces coelicolor

Pan

Liu

2021

Microbial Biotechnology

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Summary Heterologous expression of the biosynthetic gene cluster (BGC) is important for studying the microbial natural products (NPs), especially for those kept in silent or poorly expressed in their original strains. Here, we cloned the spinosad BGC through the Cas9‐Assisted Targeting of Chromosome segments and amplified it to five copies through a ZouA‐dependent DNA amplification system in Streptomyces coelicolor M1146. The resulting strain produced 1253.9 ± 78.2 μg l −1 of spinosad, which was about 224‐fold compared with that of the parent strain carrying only one copy of the spinosad BGC. Moreover, we further increased spinosad to 1958.9 ± 73.5 μg l −1 by the dynamic regulation of intracellular triacylglycerol degradation. Our study indicates that tandem amplification of the targeted gene cluster is particularly suitable to enhance the heterologous production of valuable NPs with efficiency and simplicity.

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Recombinational cloning of the antibiotic biosynthetic gene clusters in linear plasmid SCP1 ofStreptomyces coelicolorA3(2)

Cited by 6 publications

References 52 publications

Interrogation of Streptomyces avermitilis for efficient production of avermectins

Interrogation of Streptomyces avermitilis for efficient production of avermectins

Heterologous Biosynthesis of Spinosad: An Omics-Guided Large Polyketide Synthase Gene Cluster Reconstitution in Streptomyces

Multiplying the heterologous production of spinosad through tandem amplification of its biosynthetic gene cluster in Streptomyces coelicolor

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