Negative regulation of bleomycins biosynthesis by ArsR/SmtB family repressor BlmR in Streptomyces verticillus

  • Hong Chen
  • Junhua Wang
  • Jiaqi Cui
  • Cheng Wang
  • Shaoxiong Liang
  • Huanhuan Liu
  • Jianping WenEmail author
Applied genetics and molecular biotechnology


Bleomycin, a broad-spectrum antibiotic, has been widely used for various tumor treatments. However, its poor fermentation yield is not satisfactory for industrial production. Here, the ArsR/SmtB family regulator BlmR was characterized as a repressor of bleomycin production. As an autoregulator, BlmR was found to bind to a 12-2-12 imperfect palindrome sequence in its own promoter, and deletion of blmR led to a 34% increase of bleomycin B2 production compared with the wild-type strain. Using reverse transcription and quantitative PCR (RT-qPCR), blmT, which encoded a putative transporter, was identified as the target gene regulated by BlmR. Therefore, high-production strain was constructed by blmT overexpression in a blmR deletion strain, and the bleomycin B2 titer reached to 80 mg/L, which was 1.9-fold higher than the wild-type strain. Moreover, electrophoretic mobility shift assay (EMSA) showed neither metal-binding motifs nor redox switches in BlmR. In order to elucidate the regulatory mechanism, a model of BlmR was constructed by homology modeling and protein-protein docking. The BlmR-DNA complex was generated by protein-DNA docking with the assistance of site-directed mutagenesis and molecular dynamic (MD) simulation, which directly revealed several key amino acid residues needed for the maintenance and stabilization of the interface between BlmR and target DNA. The interface information could provide the configuration reference and seek the potential effectors that could interact with BlmR, thereby extending the regulation role of ArsR/SmtB family members on the improvement of antibiotic production.


Bleomycins ArsR/SmtB family regulator Molecular dynamic simulation Key amino acid residues 


Funding information

This work was financially supported by the National Natural Science Foundation of China (No. 21676189) and the Key Technologies R&D Program of Tianjin (No. 16YFZCSY00780).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2019_9923_MOESM1_ESM.pdf (708 kb)
ESM 1 (PDF 707 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Hong Chen
    • 1
    • 2
  • Junhua Wang
    • 1
  • Jiaqi Cui
    • 1
    • 2
  • Cheng Wang
    • 3
  • Shaoxiong Liang
    • 4
  • Huanhuan Liu
    • 5
  • Jianping Wen
    • 1
    • 2
    Email author
  1. 1.Key Laboratory of Systems Bioengineering (Ministry of Education)Tianjin UniversityTianjinPR China
  2. 2.SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and TechnologyTianjin UniversityTianjinPR China
  3. 3.Department of Forestry Engineering, College of ForestryNorthwest A&F UniversityYanglingPR China
  4. 4.College of Chemical EngineeringHuaqiao UniversityXiamenPR China
  5. 5.Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and BiotechnologyTianjin University of Science and TechnologyTianjinChina

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