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Characterization of 17β-hydroxysteroid dehydrogenase and regulators involved in estrogen degradation in Pseudomonas putida SJTE-1

  • Pingping Wang
  • Daning Zheng
  • Wanli Peng
  • Yanqiu Wang
  • Xiuli Wang
  • Weiliang Xiong
  • Rubing LiangEmail author
Environmental biotechnology

Abstract

In bacteria, the enzyme catalyzing the transformation of 17β-estradiol is considered the key enzyme for its metabolism, whose enzymatic activity and regulatory network influence the biodegradation efficiency of this typical estrogen. In this work, a novel 17β-hydroxysteroid dehydrogenase (17β-HSD) was characterized from the estrogen-degrading strain Pseudomonas putida SJTE-1, and two regulators were identified. This 17β-HSD, a member of the short-chain dehydrogenase/reductase (SDR) superfamily, could be induced by 17β-estradiol and catalyzed the oxidization reaction at the C17 site of 17β-estradiol efficiently. Its Km value was 0.068 mM, and its Vmax value was 56.26 μmol/min/mg; over 98% of 17β-estradiol was oxidized into estrone in 5 min, indicating higher efficiency than other reported bacterial 17β-HSDs. Furthermore, two genes (crgA and oxyR) adjacent to 17β-hsd were studied which encoded the potential CrgA and OxyR regulators. Overexpression of crgA could enhance the transcription of 17β-hsd, while that of oxyR resulted in the opposite effect. They could bind to the specific and different sites in the promoter region of 17β-hsd gene directly, and binding of OxyR could be released by 17β-estradiol. OxyR repressed the expression of 17β-hsd by its specific binding to the conserved motif of GATA-N9-TATC, while CrgA activated the expression of this gene through its binding to the motif of T-N11-A. Therefore, this 17β-HSD transformed 17β-estradiol efficiently and the two regulators regulated its expression directly. This work could promote the study of the enzymatic mechanism and regulatory network of the estrogen biodegradation pathway in bacteria.

Keywords

17β-hydroxysteroid dehydrogenase 17β-estradiol Estrone OxyR CrgA Biodegradation 

Notes

Funding

This work was supported by the National Science Foundation of China (Grant No. 31370152, 31570099) and the Shanghai Pujiang Program (14PJD020).

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.

Ethical approval

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

Supplementary material

253_2018_9543_MOESM1_ESM.pdf (166 kb)
ESM 1 (PDF 165 kb)

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Copyright information

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

Authors and Affiliations

  • Pingping Wang
    • 1
  • Daning Zheng
    • 1
  • Wanli Peng
    • 1
  • Yanqiu Wang
    • 1
  • Xiuli Wang
    • 1
  • Weiliang Xiong
    • 1
  • Rubing Liang
    • 1
    Email author
  1. 1.State Key Laboratory of Microbial Metabolism, and School of Life Sciences and BiotechnologyShanghai Jiao Tong UniversityShanghaiChina

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