ToyA, a positive pathway-specific regulator for toyocamycin biosynthesis in Streptomyces diastatochromogenes 1628

Xu, Jie; Song, Zhangqing; Xu, Xianhao; Ma, Zheng; Bechthold, Andreas; Yu, Xiaoping

doi:10.1007/s00253-019-09959-w

Cited by 21 publications

(18 citation statements)

References 47 publications

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“…This result confirmed that NsdA sr is not only a homologue of NsdA sc but also has similar effects. Subsequently, by heterologous expression of nsdA sr , similar negative effects on morphological differentiation and antibiotic production were observed in S. diastatochromogenes 1628, which is considered a significant industrial producer of the nucleoside antibiotic, TM [35,36].…”

Section: Discussionmentioning

confidence: 82%

“…Extraction of RNA and analysis of transcriptional levels of rim genes were performed as described previously [17,36]. Extraction of RNA, design of primers, and analysis of the transcription of toy genes were performed as described by Xu et al [35]. Extraction of RNA, design of primers, and analysis of transcription of actII-orf4 gene were performed as described by Zhao et al [38].…”

Section: Analysis Of Gene Transcriptional Levels By Qrt-pcrmentioning

confidence: 99%

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Identification of a gene from Streptomyces rimosus M527 negatively affecting antibiotic biosynthesis and morphological differentiation

Song

et al. 2020

Preprint

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Background: The polyene macrolide rimocidin, produced by Streptomyces rimosus M527, was found to be highly effective against a broad range of fungal plant pathogens. Current understanding of the regulatory mechanism of rimocidin biosynthesis and morphological differentiation in S. rimosus M527 is limited. NsdA is considered as a negative regulator involved in morphological differentiation and biosynthesis of secondary metabolites in some Streptomyces.Results: In this study, nsdAsr was cloned from S. rimosus M527. The role of nsdAsr in rimocidin biosynthesis and morphological differentiation was investigated by gene deletion, complementation, and over-expression. A ΔnsdAsr mutant was obtained using CRISPR/Cas9. The mutant produced more rimocidin (46%) and generated more spores than the wild-type strain. Over-expression of nsdAsr led to a decrease in rimocidin production and impairment of sporulation. Quantitative reverse transcription-PCR (qRT-PCR) analysis revealed that transcription of rim genes responsible for rimocidin biosynthesis was up-regulated in the ΔnsdAsr mutant but down-regulated in the nsdAsr over-expression strain. Similar effects have been described for Streptomyces coelicolor M145 and the industrial toyocamycin-producing strain Streptomyces diastatochromogenes 1628.Conclusion: NsdAsr is identified as a negative regulator of sporluation and antibiotic biosynthesis as well as the transcription of biosynthetic genes both in its host S. rimosus M527 and in model strain S. coelicolor and industrial producer strain S. diastatochromogenes 1628. This work will provide further information for understanding regulatory mechanisms controlling rimocidin biosynthesis in S. rimosus M527.

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Section: Discussionmentioning

confidence: 82%

Section: Analysis Of Gene Transcriptional Levels By Qrt-pcrmentioning

confidence: 99%

Identification of a gene from Streptomyces rimosus M527 negatively affecting antibiotic biosynthesis and morphological differentiation

Song

et al. 2020

Preprint

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“…The counterpart of the gene huiA within the toyocamycin biosynthetic pathway, toyA , also constitutes the first gene of the cluster [ 28 ]. The deletion of the toyA gene completely abolishes toyocamycin production [ 29 ]. The first three genes in the region upstream of the huiA gene, KALB_4064, KALB_4065, and KALB_4066 ( Figure 3 ), encode an enhanced intracellular survival protein and two hypothetical proteins, respectively.…”

Section: Resultsmentioning

confidence: 99%

Identification of a Biosynthetic Gene Cluster Responsible for the Production of a New Pyrrolopyrimidine Natural Product—Huimycin

et al. 2020

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Pyrrolopyrimidines are an important class of natural products with a broad spectrum of biological activities, including antibacterial, antifungal, antiviral, anticancer or anti-inflammatory. Here, we present the identification of a biosynthetic gene cluster from the rare actinomycete strain Kutzneria albida DSM 43870, which leads to the production of huimycin, a new member of the pyrrolopyrimidine family of compounds. The huimycin gene cluster was successfully expressed in the heterologous host strain Streptomyces albus Del14. The compound was purified, and its structure was elucidated by means of nuclear magnetic resonance spectroscopy. The minimal huimycin gene cluster was identified through sequence analysis and a series of gene deletion experiments. A model for huimycin biosynthesis is also proposed in this paper.

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“…Expression of an extra copy of toyG and frr led to a 65.5% increase in TM production ( Tao et al, 2016 ). In another study, expression of an extra copy of pathway-specific regulator toyA driven by different promoters led to up to twofold increase in TM production ( Xu et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

“…TM is produced by Streptomyces diastatochromogenes 1628, but the production level is always very low ( Ma et al, 2014a ). In this context, efforts to improve TM production have been made by the overexpression of structural genes ( Tao et al, 2016 ) or favorable genes ( Ma et al, 2014a ; Ma et al, 2014b ) and the pathway-specific regulatory gene toyA ( Xu et al, 2019 ) or ribosome engineering technology ( Ma et al, 2016 ). Expression of an extra copy of toyG and frr led to a 65.5% increase in TM production ( Tao et al, 2016 ).…”

Section: Introductionmentioning

confidence: 99%

Cloning and Overexpression of the Toy Cluster for Titer Improvement of Toyocamycin in Streptomyces diastatochromogenes

Liao

et al. 2020

Front. Microbiol.

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The nucleoside antibiotic toyocamycin (TM) is a potential fungicide that can control plant diseases, and it has become an attractive target for research. Streptomyces diastatochromogenes 1628, a TM-producing strain, was isolated by our laboratory and was considered to be a potent industrial producer of TM. Recently, the putative TM biosynthetic gene cluster ( toy cluster) in S. diastatochromogenes 1628 was found by genome sequencing. In this study, the role of toy cluster for TM biosynthesis in S. diastatochromogenes 1628 was investigated by heterologous expression, deletion, and complementation. The extract of the recombinant strain S. albus J1074-TC harboring a copy of toy cluster produced TM as shown by HPLC analysis. The Δcluster mutant completely lost its ability to produce TM. TM production in the complemented strain was restored to a level comparable to that of the wild-type strain. These results confirmed that the toy cluster is responsible for TM biosynthesis. Moreover, the introduction of an extra copy of the toy cluster into S. diastatochromogenes 1628 led to onefold increase in TM production (312.9 mg/l vs. 152.1 mg/l) as well as the transcription of all toy genes. The toy gene cluster was engineered in which the native promoter of toyA gene, toyM gene, toyBD operon, and toyEI operon was, respectively, replaced by permE ∗ or SPL57. To further improve TM production, the engineered toy gene cluster was, respectively, introduced and overexpressed in S. diastatochromogenes 1628 to generate recombinant strains S. diastatochromogenes 1628-EC and 1628-SC. After 84 h, S. diastatochromogenes 1628-EC and 1628-SC produced 456.5 mg/l and 638.9 mg/l TM, respectively, which is an increase of 2- and 3.2-fold compared with the wild-type strain.

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ToyA, a positive pathway-specific regulator for toyocamycin biosynthesis in Streptomyces diastatochromogenes 1628

Cited by 21 publications

References 47 publications

Identification of a gene from Streptomyces rimosus M527 negatively affecting antibiotic biosynthesis and morphological differentiation

Identification of a gene from Streptomyces rimosus M527 negatively affecting antibiotic biosynthesis and morphological differentiation

Identification of a Biosynthetic Gene Cluster Responsible for the Production of a New Pyrrolopyrimidine Natural Product—Huimycin

Cloning and Overexpression of the Toy Cluster for Titer Improvement of Toyocamycin in Streptomyces diastatochromogenes

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