FadR1, a pathway-specific activator of fidaxomicin biosynthesis in Actinoplanes deccanensis Yp-1

Liu, Yueping; Yu, Ping; Li, Jifeng; Yu-qi, Tang; Bu, Qing-Ting; Mao, Xu‐Ming; Li, Yongquan

doi:10.1007/s00253-019-09949-y

Cited by 11 publications

(12 citation statements)

References 50 publications

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“…In our hands, D. aurantiacum was found to be very difficult to cultivate. Instead, we turned to the genetically tractable actinobacterium A. deccanensis [26–28] . Previous studies and patents describe cultivation of A. deccanensis and genetically engineered A. deccanensis strains, resulting in Fdx titers of 65.6 mg/L (not isolated), [29] 130 mg/L (not isolated), [30] 619 mg/L (not isolated), [28] 900 mg/L (not isolated), [31] and >1500 mg/L (crude) [32] .…”

Section: Resultsmentioning

confidence: 99%

Isolation of Fidaxomicin and Shunt Metabolites from Actinoplanes deccanensis

Jung,

Hunter,

Dorst

et al. 2024

Helvetica Chimica Acta

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A protocol for the isolation of the antibiotic fidaxomicin (Fdx) from Actinoplanes deccanensis and the isolation of shunt metabolites from A. deccanensisfdxG2‐ is reported. We constructed the mutant strain A. deccanensisfdxG2‐ by genetic manipulation which enabled the isolation of shunt metabolites as useful starting points for semisynthetic analogues of Fdx. Furthermore, a synthetic protocol for the conversion of complex A. deccanensisfdxG2‐ extracts into the single compound FdxG2‐OH via methanolysis is presented. This synthetic procedure is complemented by images and practical notes. Full structure assignment is given in the SI and the characterization data files are published to aid experimentalists. The protocol is also suitable as an undergraduate laboratory project. We hope to facilitate research into new Fdx derivatives through the availability of this procedure.

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Section: Resultsmentioning

confidence: 99%

Isolation of Fidaxomicin and Shunt Metabolites from Actinoplanes deccanensis

Jung,

Hunter,

Dorst

et al. 2024

Helvetica Chimica Acta

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“…The 5′-(6-FAM)-labeled mutant probe was designed based on the results of EMSA and footprinting results and generated by PCR with the primers listed in Table S3. Each binding reaction contained 0.5 ng of the probe and increasing concentrations of TagR as indicated ( 67 ).…”

Section: Methodsmentioning

confidence: 99%

“…The 5′-(6-FAM)-labeled DNA probes for the DNase I footprinting assay were generated from the plasmid pKC1139- P tagR using the primer tagGR -P-F(5′-FAM)/ tagGR -P-R. The DNase I footprinting assay was carried out as previously described ( 67 ).…”

Section: Methodsmentioning

confidence: 99%

Identification and Characterization of a New Regulator, TagR, for Environmental Stress Resistance Based on the DNA Methylome of Streptomyces roseosporus

Gao,

Fang,

Zhu

et al. 2023

Microbiol Spectr

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This study established a novel strategy for screening regulators of environmental stress resistance based on the DNA methylome and discovered a new regulator, TagR. The TagR-WTA transporter regulatory pathway improved the resistance and antibiotic yield of strains and has the potential for wide application. Our research provides a new perspective on the optimization and reconstruction of industrial actinomycetes.

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“…AmphRIV, which regulates amphotericin biosynthesis in Streptomyces nodosus [ 26 ], and NysRIV, which regulates nystatin biosynthesis in Streptomyces noursei [ 27 ]. (3) Larger ATP-binding regulators of the LuxR (LAL) family regulators, which are characterized by an unusually large number of amino acids (~ 900) [ 28 ] having an ATP-binding motif near the N-terminal end and Walker A, Walker B, and a HTH domain at the C-terminus. Numerous regulators belonging to the LAL family have been identified, such as TtmRIII, which regulates tetramycin biosynthesis in Streptomyces ahygroscopicus [ 29 ], and NysRI, NysRII, and NysRIII, which regulates nystatin biosynthesis in S. noursei ATCC 11,455 [ 30 ].…”

Section: Introductionmentioning

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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FadR1, a pathway-specific activator of fidaxomicin biosynthesis in Actinoplanes deccanensis Yp-1

Cited by 11 publications

References 50 publications

Isolation of Fidaxomicin and Shunt Metabolites from Actinoplanes deccanensis

Isolation of Fidaxomicin and Shunt Metabolites from Actinoplanes deccanensis

Identification and Characterization of a New Regulator, TagR, for Environmental Stress Resistance Based on the DNA Methylome of Streptomyces roseosporus

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

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