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Natural Disinfection-like Process Unveiled in Soil Microenvironments by Enzyme-Catalyzed Chlorination.

Xiangyu Zhu1,2, Kun Wang1,3, Congcong Liu1,2

  • 1Department of Environmental Science, Zhejiang University, Hangzhou, Zhejiang 310058, China.

Environmental Science & Technology
|February 14, 2024
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Natural enzymatic chlorination, driven by heme-containing chloroperoxidase (CPO), effectively disinfects soils by producing reactive chlorine species (RCS). This process shows enhanced antibacterial activity against soil bacteria like Escherichia coli and Bacillus subtilis.

Keywords:
disinfection byproductmicrofluidic chipnatural enzymatic chlorinationreactive chlorine speciesreactive oxygen speciessoil organic matter

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Area of Science:

  • Environmental microbiology
  • Biogeochemistry
  • Soil science

Background:

  • Natural chlorination processes in terrestrial ecosystems are significant, involving abiotic and biological pathways.
  • Exoenzymatically mediated chlorination is a key route for organochlorine formation and chloride conversion to reactive chlorine species (RCS).
  • The antibacterial role of natural enzymatic chlorination in soil microenvironments is largely unknown.

Purpose of the Study:

  • To investigate the potential of heme-containing chloroperoxidase (CPO)-catalyzed chlorination as a natural soil disinfection mechanism.
  • To evaluate the antibacterial activity of enzymatic chlorination against common soil bacteria.
  • To elucidate the role of reactive chlorine species (RCS) in this natural disinfection process.

Main Methods:

  • Antimicrobial experiments were conducted to assess antibacterial activity.
  • Microfluidic chip-based fluorescence imaging was used to visualize RCS distribution and effective antibacterial range.
  • Ultrahigh-resolution mass spectrometry analyzed chlorine-containing formulas from CPO-catalyzed and abiotic chlorination.

Main Results:

  • Enzymatic chlorination demonstrated significantly enhanced antibacterial activity against Escherichia coli and Bacillus subtilis compared to hydrogen peroxide alone.
  • In situ-formed RCS were identified as the primary drivers of the enhanced antibacterial effect.
  • The effective antibacterial distance of CPO-catalyzed chlorination was estimated to be approximately 2 mm.
  • Chlorine-containing formulas from CPO-catalyzed chlorination closely resembled those from abiotic chlorination of dissolved organic matter.

Conclusions:

  • Heme-containing chloroperoxidase (CPO) catalyzes a natural soil disinfection process.
  • This enzymatic process effectively utilizes reactive chlorine species (RCS) for antibacterial activity.
  • Findings link chlorine-carbon interactions and reactive species dynamics in natural soil environments, revealing a novel disinfection pathway.