Novel Aflatoxin-Degrading Enzyme from Bacillus shackletonii L7

Toxins (Basel). 2017 Jan 14;9(1):36. doi: 10.3390/toxins9010036.

Abstract

Food and feed contamination by aflatoxin (AF)B₁ has adverse economic and health consequences. AFB₁ degradation by microorganisms or microbial enzymes provides a promising preventive measure. To this end, the present study tested 43 bacterial isolates collected from maize, rice, and soil samples for AFB₁-reducing activity. The higher activity was detected in isolate L7, which was identified as Bacillus shackletonii. L7 reduced AFB₁, AFB₂, and AFM₁ levels by 92.1%, 84.1%, and 90.4%, respectively, after 72 h at 37 °C. The L7 culture supernatant degraded more AFB₁ than viable cells and cell extracts; and the degradation activity was reduced from 77.9% to 15.3% in the presence of proteinase K and sodium dodecyl sulphate. A thermostable enzyme purified from the boiled supernatant was designated as Bacillus aflatoxin-degrading enzyme (BADE). An overall 9.55-fold purification of BADE with a recovery of 39.92% and an activity of 3.85 × 10³ U·mg-1 was obtained using chromatography on DEAE-Sepharose. BADE had an estimated molecular mass of 22 kDa and exhibited the highest activity at 70 °C and pH 8.0, which was enhanced by Cu2+ and inhibited by Zn2+, Mn2+, Mg2+, and Li⁺. BADE is the major protein involved in AFB₁ detoxification. This is the first report of a BADE isolated from B. shackletonii, which has potential applications in the detoxification of aflatoxins during food and feed processing.

Keywords: Bacillus shackletonii; aflatoxin B1; aflatoxin-degrading enzyme; biodegradation; purification.

MeSH terms

  • Aflatoxin B1 / metabolism*
  • Aflatoxin M1 / metabolism*
  • Aflatoxins / metabolism*
  • Bacillus / classification
  • Bacillus / enzymology*
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism*
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Inactivation, Metabolic
  • Metals / chemistry
  • Substrate Specificity
  • Temperature
  • Time Factors

Substances

  • Aflatoxins
  • Bacterial Proteins
  • Metals
  • Aflatoxin M1
  • aflatoxin B2
  • Aflatoxin B1