Impact of otrA expression on morphological differentiation, actinorhodin production, and resistance to aminoglycosides in Streptomyces coelicolor M145

Zhao, Ying; Lu, Dandan; Bechthold, Andreas; Ma, Zheng; Yu, Xiaoping

doi:10.1631/jzus.b1800046

Cited by 9 publications

(5 citation statements)

References 33 publications

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“…Media and culture conditions of Escherichia coli, S. rimosus M527 and its derivates were described by Zhao et al (2018). OTC fermentation medium (starch 30 g/l, soya flour 3 g/l, (NH 3 ) 2 SO 4 4 g/l, CaCO 3 5 g/l, NaCl 5 g/l, corn steep liquor 5 g/l, K 2 HPO 4 0.5 g/l, CoCl 2 0.01 g/l, tap water) in a 250‐mL Erlenmeyer flask.…”

Section: Methodsmentioning

confidence: 99%

Double‐reporter‐guided targeted activation of the oxytetracycline silent gene cluster in Streptomyces rimosus M527

Shi

Zhang

et al. 2023

Biotech & Bioengineering

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In Streptomyces rimosus M527, the oxytetracycline (OTC) biosynthetic gene cluster is not expressed under laboratory conditions. In this study a reported‐guided mutant selection (RGMS) procedure was used to activate the cluster. The double‐reporter plasmid pAGT was constructed in which gusA encoding a β‐glucuronidase and tsr encoding a thiostrepton resistance methyltransferase were placed under the control of the native promoter of oxyA gene (PoxyA). Plasmid pAGT was introduced and integrated into the chromosome of S. rimosus M527 by conjugation, yielding initial strain M527‐pAGT. Subsequently, mutants of M527‐pAGT were generated by using ribosome engineering technology. The mutants harboring activated OTC gene cluster were selected based on visual observation of GUS activity and thiostrepton resistance. Finally, mutant M527‐pAGT‐R7 was selected producing OTC in a concentration of 235.2 mg/L. In this mutant transcriptional levels of oxysr genes especial oxyAsr gene were increased compared to wild‐type strain S. rimosus M527. The mutant M527‐pAGT‐R7 showed antagonistic activities against Gram‐negative and Gram‐positive strains. All data indicate that the OTC gene cluster was successfully activated using the RGMS method.

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Section: Methodsmentioning

confidence: 99%

Double‐reporter‐guided targeted activation of the oxytetracycline silent gene cluster in Streptomyces rimosus M527

Shi

Zhang

et al. 2023

Biotech & Bioengineering

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“…The pib139-EGFP plasmid was transformed into strain Z1-26 using the method of conjugal transfer (Zhao et al, 2018). The growth and fluorescence analysis of strain Z1-26 EGFP were monitored.…”

Section: Colonization and Distribution Of Z1-26 Egfp Strain In Crucia...mentioning

confidence: 99%

A new isolate of Streptomyces lateritius (Z1‐26) with antibacterial activity against fish pathogens and immune enhancement effects on crucian carp (Carassius auratus)

Yang

Jin

Long

et al. 2022

Journal of Fish Diseases

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The Streptomyces lateritius Z1-26 was isolated from soil samples which showed broadspectrum antibacterial activity against a broad range of fish pathogens. The In Vivo Imaging System (IVIS) monitored that strain Z1-26 could survive and colonize in the gills and abdomen of crucian carp. The effects of dietary supplementation with strain Z1-26 were evaluated with respect to the growth performance, antioxidant capacity, and immune response of crucian carp. The results showed that the Z1-26-fed fish had a significantly higher growth rate than the fish fed the control diet. The immune and antioxidant parameters revealed that the non-specific immune indicators (AKP, SOD, and LZM) of the serum, the expression of immune-related genes (IgM, C3, and LZM), and antioxidant-related genes (Nrf2 and Keap1) of the immune organs were significantly increased, whereas the expression of pro-inflammatory factors (IL-1β, IL-8, and TNFα) of the immune organs was significantly down-regulated in crucian carp fed strain Z1-26 compared with fish fed a control diet. Moreover, fish fed with Z1-26 supplemented diets showed a significantly improved survival rate after Aeromonas hydrophila infection. In addition, the whole genome analysis showed that strain Z1-26 possesses 28 gene clusters, including 6 polyketide synthetase (PKS), 4 non-ribosomal peptide-synthetase (NRPS), 1 bacteriocin, and 1 lantipeptide. In summary, these results indicated that strain Z1-26 could improve the growth performance and disease resistance in crucian carp, and has the potential to be developed as a candidate probiotics for the control of bacterial diseases in aquaculture.

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“…2CMC solid medium [16] was used for conjugation. CP liquid medium [38] was used as seed medium. S. rimosus M527 and its derivates were incubated using the method of Zhao et al [17].…”

Section: Strains Plasmids and Primersmentioning

confidence: 99%

“…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%

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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Impact of otrA expression on morphological differentiation, actinorhodin production, and resistance to aminoglycosides in Streptomyces coelicolor M145

Cited by 9 publications

References 33 publications

Double‐reporter‐guided targeted activation of the oxytetracycline silent gene cluster in Streptomyces rimosus M527

Double‐reporter‐guided targeted activation of the oxytetracycline silent gene cluster in Streptomyces rimosus M527

A new isolate of Streptomyces lateritius (Z1‐26) with antibacterial activity against fish pathogens and immune enhancement effects on crucian carp (Carassius auratus)

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

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