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

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.

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Funding

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

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Correspondence to Rubing Liang.

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Wang, P., Zheng, D., Peng, W. et al. Characterization of 17β-hydroxysteroid dehydrogenase and regulators involved in estrogen degradation in Pseudomonas putida SJTE-1. Appl Microbiol Biotechnol 103, 2413–2425 (2019). https://doi.org/10.1007/s00253-018-9543-y

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  • DOI: https://doi.org/10.1007/s00253-018-9543-y

Keywords

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