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Isolation and purification of an enzyme hydrolyzing ochratoxin A from <Emphasis Type="Italic">Aspergillus niger</Emphasis>
Authors:Luís Abrunhosa  Armando Venâncio
Institution:(1) IBB- Institute for Biotechnology and Bioengineering, Centre for Biological Engineering, Universidade do Minho, Campus de Gualtar, Brague, 4710-057, Portugal;(2) Departamento de Engenharia Biológica, Universidade do Minho, Campus de Gualtar, Brague, 4710-057, Portugal
Abstract:Ochratoxin A is a mycotoxin produced by several Aspergillus and some Penicillium species which may be present in food and feed products. It can be enzymatically hydrolyzed into ochratoxin α and l-β-phenylalanine, thereby decreasing its toxicity. The ochratoxin A degradation capacity of Aspergillus niger is well known and here we report the isolation and purification of a novel enzyme from A. niger that hydrolyzes this mycotoxin. A wheat germ medium supplemented with ochratoxin A was used to produce the enzyme, which was purified from culture filtrate by acetone precipitation and anion exchange chromatography. An overall purification of 2.5-fold with a recovery of 68% and a final specific activity of 36 U/mg was obtained. The enzyme is a metalloenzyme as it was inhibited at 10 mM EDTA, whereas PMSF had no effect. The ochratoxin A hydrolytic enzyme presented a V max of 0.44 μM/min and a K m of 0.5 mM when the reaction was carried out at pH 7.5 and 37°C.
Keywords:Aspergillus           niger            Carboxypeptidase  Degradation  Ochratoxin A  Ochratoxin α  
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