Heterologous expression of the tetracycline resistance gene tetX to enhance degradability and safety in doxycycline degradation

Wen X 1 , Huang J 1 , Cao J 1 , Xu J 1 , Mi J 2 , Wang Y 2 Show all authors , Ma B 3 , Zou Y 3 , Liao X 2 , Liang JB 4 , Wu Y 5

Affiliations 

  • 1 Guangdong Laboratory for Lingnan Modern Agriculture, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China; National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China; Ministry of Agriculture Key Laboratory of Tropical Agricultural Environment, South China Agricultural University, Guangzhou, 510642, China
  • 2 Guangdong Laboratory for Lingnan Modern Agriculture, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China; National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China; Ministry of Agriculture Key Laboratory of Tropical Agricultural Environment, South China Agricultural University, Guangzhou, 510642, China; Guangdong Enterprise Lab of Healthy Animal Husbandry and Environment Control, Yunfu, Xinxing, 527400, China
  • 3 Nanhai Office of Foshan Customs House, Foshan, 528200, China
  • 4 Laboratory of Animal Production, Institute of Tropical Agriculture, Universiti Putra Malaysia, Serdang, 43400, Malaysia
  • 5 Guangdong Laboratory for Lingnan Modern Agriculture, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China; National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China; Ministry of Agriculture Key Laboratory of Tropical Agricultural Environment, South China Agricultural University, Guangzhou, 510642, China; Guangdong Enterprise Lab of Healthy Animal Husbandry and Environment Control, Yunfu, Xinxing, 527400, China. Electronic address: [email protected]
Ecotoxicol Environ Saf, 2020 Mar 15;191:110214.
PMID: 31968275 DOI: 10.1016/j.ecoenv.2020.110214

Abstract

Microbial remediation has the potential to inexpensively yet effectively decontaminate and restore contaminated environments, but the virulence of pathogens and risk of resistance gene transmission by microorganisms during antibiotic removal often limit its implementation. Here, a cloned tetX gene with clear evolutionary history was expressed to explore doxycycline (DOX) degradation and resistance variation during the degradation process. Phylogenetic analysis of tetX genes showed high similarity with those of pathogenic bacteria, such as Riemerella sp. and Acinetobacter sp. Successful tetX expression was performed in Escherichia coli and confirmed by SDS-PAGE and Western blot. Our results showed that 95.0 ± 1.0% of the DOX (50 mg/L) was degraded by the recombinant strain (ETD-1 with tetX) within 48 h, which was significantly higher than that for the control (38.9 ± 8.7%) and the empty plasmid bacteria (8.8 ± 5.1%) (P  0.05). The efficient and safe DOX-degrading capacity of the recombinant strain ETD-1 makes it valuable and promising for antibiotic removal in the environment.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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