A novel lactonohydrolase responsible for the detoxification of zearalenone: enzyme purification and gene cloning.
Zearalenone (ZEN) is converted into a far less oestrogenic product by incubation with Clonostachys rosea IFO 7063. An alkaline hydrolase responsible for the detoxification was purified to homogeneity from the fungus by a combination of salt precipitation and column chromatography methods. The purified enzyme was homodimeric with a subunit molecular mass of 30 kDa and contained an intra-subunit disulphide bridge. On the basis of the internal peptide sequences of the purified protein, we cloned the entire coding region of the gene (designated as zhd101) by PCR techniques. The ZEN degradation activity was detected in heterologous hosts (Schizosaccharomyces pombe and Escherichia coli) carrying the cloned gene. Zhd101 could be a promising genetic resource for in planta detoxification of the mycotoxin in important crops.
Takahashi-Ando
Naoko
N
Laboratory for Remediation Research, Plant Science Centre, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Kimura
Makoto
M
Kakeya
Hideaki
H
Osada
Hiroyuki
H
Yamaguchi
Isamu
I
D000595:Amino Acid Sequence; D001483:Base Sequence; D001673:Biodegradation, Environmental; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D004968:Estrogens, Non-Steroidal / Q000378:metabolism; D005656:Fungal Proteins; D015870:Gene Expression; D005800:Genes, Fungal; D020462:Gliocladium / Q000201:enzymology* / Q000235:genetics*; D006867:Hydrolases / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D008658:Inactivation, Metabolic; D008969:Molecular Sequence Data; D009183:Mycotoxins / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D012568:Schizosaccharomyces / Q000235:genetics; D015025:Zearalenone / Q000378:metabolism*
D004271:DNA, Fungal; D004968:Estrogens, Non-Steroidal; D005656:Fungal Proteins; D009183:Mycotoxins; D015025:Zearalenone; D006867:Hydrolases; C460562:Zhd101 protein, Bionectria ochroleuca