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

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

doi:10.1007/s00253-020-10955-8

Cited by 12 publications

(8 citation statements)

References 44 publications

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“…Several recent studies focused on the production of oxytetracycline, a broad-spectrum antibiotic, by Streptomyces rimosus [ 8 , 9 , 10 , 11 ]. This species is also known to secrete an antifungal substance rimocidin [ 12 , 13 ] and harbors the genetic basis for the desferrioxamine siderophore biosynthesis [ 14 ]. In a recent study, Kuhl et al [ 15 ] demonstrated that inoculating actinobacteria into the liquid medium containing talc microparticles resulted in morphological and production-related alterations compared to the conventional cultures.…”

Section: Introductionmentioning

confidence: 99%

Enhanced Oxytetracycline Production by Streptomyces rimosus in Submerged Co-Cultures with Streptomyces noursei

Boruta

Ścigaczewska

2021

Molecules

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In the present study, Streptomyces rimosus was confronted with Streptomyces noursei, Penicillium rubens, Aspergillus niger, Chaetomium globosum, or Mucor racemosus in two-species submerged co-cultures in shake flasks with the goal of evaluating the oxytetracycline production and morphological development. The co-culture of S. rimosus with S. noursei exhibited stimulation in oxytetracycline biosynthesis compared with the S. rimosus monoculture, whereas the presence of M. racemosus resulted in a delay in antibiotic production. Different strategies of initiating the “S. rimosus + S. noursei” co-cultures were tested. The improvement in terms of oxytetracycline titers was recorded in the cases where S. noursei was co-inoculated with S. rimosus in the form of spores. As the observed morphological changes were not unique to the co-culture involving S. noursei, there was no evidence that the improvement of oxytetracycline levels could be attributed mainly to morphology-related characteristics.

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Section: Introductionmentioning

confidence: 99%

Enhanced Oxytetracycline Production by Streptomyces rimosus in Submerged Co-Cultures with Streptomyces noursei

Boruta

Ścigaczewska

2021

Molecules

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“…According to the previously described methods [ 49 , 54 ], the disruption of the rimR 2 gene was performed by using the CRISPR/Cas9. In terms of plasmid construction, the 0.3-kb sgRNA cloning cassette was inserted into pWHU2653 between Nhe I/ Xba I using an infusion cloning kit, generating plasmid pWHU2653-sgRNA.…”

Section: Methodsmentioning

confidence: 99%

“…Construction of plasmid pSET152:: rimR 2 harboring the coding region of rimR 2 gene and its 300-bp promoter sequence was presented in Additional file 6 : Figure S5. According to previous methods as described by Liao et al [ 54 ], a similar procedure was adopted and performed to generate complemented strain S. rimosus M527-ΔrimR2/pSET152:: rimR 2. As a control, the empty vector pSET152 was also introduced into mutant M527-ΔrimR2 by conjugation to generate strain M527-ΔrimR2/pSET152.…”

Section: Methodsmentioning

confidence: 99%

Identification of RimR2 as a positive pathway-specific regulator of rimocidin biosynthesis in Streptomyces rimosus M527

Zhang

et al. 2023

Microb Cell Fact

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Background Streoptomyces rimosus M527 is a producer of the polyene macrolide rimocidin which shows activity against various plant pathogenic fungi. Notably, the regulatory mechanisms underlying rimocidin biosynthesis are yet to be elucidated. Results In this study, using domain structure and amino acid alignment and phylogenetic tree construction, rimR2, which located in the rimocidin biosynthetic gene cluster, was first found and identified as a larger ATP-binding regulators of the LuxR family (LAL) subfamily regulator. The rimR2 deletion and complementation assays were conducted to explore its role. Mutant M527-ΔrimR2 lost its ability to produce rimocidin. Complementation of M527-ΔrimR2 restored rimocidin production. The five recombinant strains, M527-ER, M527-KR, M527-21R, M527-57R, and M527-NR, were constructed by overexpressing rimR2 gene using the promoters permE*, kasOp*, SPL21, SPL57, and its native promoter, respectively, to improve rimocidin production. M527-KR, M527-NR, and M527-ER exhibited 81.8%, 68.1%, and 54.5% more rimocidin production, respectively, than the wild-type (WT) strain, while recombinant strains M527-21R and M527-57R exhibited no obvious differences in rimocidin production compared with the WT strain. RT-PCR assays revealed that the transcriptional levels of the rim genes were consistent with the changes in rimocidin production in the recombinant strains. Using electrophoretic mobility shift assays, we confirmed that RimR2 can bind to the promoter regions of rimA and rimC. Conclusion A LAL regulator RimR2 was identified as a positive specific-pathway regulator of rimocidin biosynthesis in M527. RimR2 regulates the rimocidin biosynthesis by influencing the transcriptional levels of rim genes and binding to the promoter regions of rimA and rimC.

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“…The extraction of RNA and analysis of transcriptional levels of rimA and neo genes in the WT strain M527, the initial strain M527-pAN and three mutants M527-pAN-S34, S38, and S52 were performed as described by Liao et al. ( 2020 ).…”

Section: Methodsmentioning

confidence: 99%

Improvement of rimocidin production in Streptomyces rimosus M527 by reporter-guided mutation selection

Jiang

Zhang

Huang

et al. 2022

Journal of Industrial Microbiology and Biotechnology

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In this study, we employed a reporter-guided mutation selection (RGMS) strategy to improve the rimocidin production of Streptomyces rimosus M527, which is based on a single-reporter plasmid pAN and atmospheric and room temperature plasma (ARTP). In plasmid pAN, PrimA, a native promoter of the loading module of rimocidin biosynthesis (RimA) was chosen as target, and the kanamycin resistance gene (neo) under the control of PrimA was chosen as reporter gene. The integrative plasmid pAN was introduced into the chromosome of S. rimosus M527 by conjugation to yield the initial strain S. rimosus M527-pAN. Subsequently, mutants of M527-pAN were generated by ARTP. Seventy-nine mutants were obtained in total, of which sixty-seven mutants showed higher level of kanamycin resistance (Kanr) than that of the initial strain M527-pAN. The majority of mutants exhibited a slight increase in rimocidin production compared to M527-pAN. Notably, three mutants, M527-pAN-S34, S38, and S52, which exhibited highest kanamycin resistance among all Kanr mutants, showed 34%, 52% and 45% increase in rimocidin production compared to M527-pAN, respectively. Quantitative RT-PCR analysis revealed that the transcriptional levels of neo and rim genes were increased in mutants M527-pAN-S34, S38, and S52 compared to M527-pAN. These results confirmed that the RGMS approach was successful in improving the rimocidin production in S. rimosus M527.

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

Cited by 12 publications

References 44 publications

Enhanced Oxytetracycline Production by Streptomyces rimosus in Submerged Co-Cultures with Streptomyces noursei

Enhanced Oxytetracycline Production by Streptomyces rimosus in Submerged Co-Cultures with Streptomyces noursei

Identification of RimR2 as a positive pathway-specific regulator of rimocidin biosynthesis in Streptomyces rimosus M527

Improvement of rimocidin production in Streptomyces rimosus M527 by reporter-guided mutation selection

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