Isolation and characterization of a Bacillus amyloliquefaciens strain with zearalenone removal ability and its probiotic potential

PLoS One. 2017 Aug 1;12(8):e0182220. doi: 10.1371/journal.pone.0182220. eCollection 2017.

Abstract

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by Fusarium species, which has been shown to be associated with reproductive disorders in livestock, and to a lesser extent with hyperoestrogenic syndromes in humans. The aim of this study was to characterize a Bacillus amyloliquefaciens strain with ZEN removal ability. A pure culture of a strain designated LN isolated from moldy corn samples showed a high ZEN removal capability. Based on microscopic observations, biochemical characteristics, and phylogenetic analysis of the 16S rRNA gene sequence, LN was identified as B. amyloliquefaciens. After incubation of B. amyloliquefaciens LN in Luria-Bertani (LB) medium containing 3.5 ppm of ZEN, the ZEN concentration fell below the detection limit within 24 h. In ZEN-contaminated corn meal medium, B. amyloliquefaciens LN decreased ZEN concentration by 92% after 36 h of incubation. In phosphate-buffered saline (PBS) containing 5 ppm of ZEN, B. amyloliquefaciens LN reduced the ZEN concentration from 5 ppm to 3.28 ppm immediately after coming into contact with ZEN, and further reduced the ZEN concentration to 0.36 ppm after 4 h of incubation. The amounts of ZEN adsorbed by the cells of B. amyloliquefaciens LN did not increase with the extension of incubation time, indicating that B. amyloliquefaciens LN not only possessed ZEN adsorption ability, but also exhibited the ability to degrade ZEN. In addition, B. amyloliquefaciens LN was non-hemolytic, non-enterotoxin producing, and displayed probiotic characteristics including acidic tolerance, bile salt tolerance, and anti-pathogenic activities. These findings suggest that B. amyloliquefaciens LN has a potential to be used as a feed additive to reduce the concentrations of ZEN in feedstuffs.

MeSH terms

  • Adsorption
  • Bacillus amyloliquefaciens / genetics
  • Bacillus amyloliquefaciens / metabolism*
  • Bile Acids and Salts / metabolism
  • Biodegradation, Environmental
  • Food Contamination*
  • Food Microbiology*
  • Humans
  • Kinetics
  • Phenotype
  • Phylogeny
  • Probiotics*
  • RNA, Ribosomal, 16S / genetics
  • Zea mays / microbiology
  • Zearalenone / metabolism*

Substances

  • Bile Acids and Salts
  • RNA, Ribosomal, 16S
  • Zearalenone

Grants and funding

This research was conducted using funds provided by National Taiwan University, grant MOST 105-2313-B-002-042-MY3 from the Ministry of Science and Technology, and grants 105AS-2.3.3-AD-U1 and 106AS-2.3.3-AD-U1 from the Council of Agriculture, Executive Yuan, Republic of China. No additional external funding was received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.