title: Secretory expression and characterization of a novel peroxiredoxin for zearalenone detoxification in Saccharomyces cerevisiae.
abstract: Zearalenone (ZEN) is a Fusarium mycotoxin, which is considered to be an oestrogenic endocrine disruptor found to cause severe morphological and functional disorders of reproductive organs in livestock. Increasing attention has been paid to the development of an effective strategy for ZEN decontamination. ZEN is oxidized into smaller estrogenic metabolites by a novel peroxiredoxin (Prx) isolated from Acinetobacter sp. SM04. The Prx coding gene was cloned in a secretory vector pYES2-alpha (pYα) with an alpha (α) signal peptide gene inserted into the multiple cloning site of pYES2. The recombinant Prx was secreted from Saccharomyces cerevisiae INVSc1 after inducing with 2% (w/v) galactose for 72 h, and was found to be nearly 20 kDa through 12% SDS-PAGE. The expressed amount of recombinant Prx was 0.24 mg/mL in the extracellular supernatant. Recombinant Prx showed a gradient increase at the beginning of ZEN degradation. The final ZEN degradation amount was 0.43 μg by one unit recombinant Prx after 12 h. Furthermore, the temperature, H(2)O(2) concentration, and pH for highest peroxidase activity of recombinant Prx were 80°C, 20 mM and 9.0, respectively. When compared with other peroxidases, the thermal stability and alkali resistance of recombinant Prx were much better. The results suggest that recombinant Prx is successfully expressed in S. cerevisiae.
authors.affiliation: College of Light Industry and Food Sciences, South China University of Technology, Guangzhou 510640, China.
authors.forename: Yuqian
authors.initials: Y
authors.lastname: Tang
authors.forename: Junmei
authors.initials: J
authors.lastname: Xiao
authors.forename: Yi
authors.initials: Y
authors.lastname: Chen
authors.forename: Yigang
authors.initials: Y
authors.lastname: Yu
authors.forename: Xinglong
authors.initials: X
authors.lastname: Xiao
authors.forename: Yuanshan
authors.initials: Y
authors.lastname: Yu
authors.forename: Hui
authors.initials: H
authors.lastname: Wu
mesh_terms: D000150:Acinetobacter / Q000201:enzymology / Q000235:genetics; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D015870:Gene Expression; D005822:Genetic Vectors; D006861:Hydrogen Peroxide / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D054464:Peroxiredoxins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D013696:Temperature; D015025:Zearalenone / Q000037:antagonists & inhibitors*
chemical_list: D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D015025:Zearalenone; D006861:Hydrogen Peroxide; D054464:Peroxiredoxins