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. 2002 Jul 1;365(Pt 1):1–6. doi: 10.1042/BJ20020450

A novel lactonohydrolase responsible for the detoxification of zearalenone: enzyme purification and gene cloning.

Naoko Takahashi-Ando 1, Makoto Kimura 1, Hideaki Kakeya 1, Hiroyuki Osada 1, Isamu Yamaguchi 1
PMCID: PMC1222652  PMID: 11978180

Abstract

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.

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Selected References

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  1. Ballance D. J. Sequences important for gene expression in filamentous fungi. Yeast. 1986 Dec;2(4):229–236. doi: 10.1002/yea.320020404. [DOI] [PubMed] [Google Scholar]
  2. Battershill J. M., Fielder R. J. Mouse-specific carcinogens: an assessment of hazard and significance for validation of short-term carcinogenicity bioassays in transgenic mice. Hum Exp Toxicol. 1998 Apr;17(4):193–205. doi: 10.1177/096032719801700401. [DOI] [PubMed] [Google Scholar]
  3. Etienne M., Jemmali M. Effects of zearalenone (F2) on estrous activity and reproduction in gilts. J Anim Sci. 1982 Jul;55(1):1–10. doi: 10.2527/jas1982.5511. [DOI] [PubMed] [Google Scholar]
  4. Kimura M., Kamakura T., Tao Q. Z., Kaneko I., Yamaguchi I. Cloning of the blasticidin S deaminase gene (BSD) from Aspergillus terreus and its use as a selectable marker for Schizosaccharomyces pombe and Pyricularia oryzae. Mol Gen Genet. 1994 Jan;242(2):121–129. doi: 10.1007/BF00391004. [DOI] [PubMed] [Google Scholar]
  5. Kimura M., Kaneko I., Komiyama M., Takatsuki A., Koshino H., Yoneyama K., Yamaguchi I. Trichothecene 3-O-acetyltransferase protects both the producing organism and transformed yeast from related mycotoxins. Cloning and characterization of Tri101. J Biol Chem. 1998 Jan 16;273(3):1654–1661. doi: 10.1074/jbc.273.3.1654. [DOI] [PubMed] [Google Scholar]
  6. Kimura M., Shingu Y., Yoneyama K., Yamaguchi I. Features of Tri101, the trichothecene 3-O-acetyltransferase gene, related to the self-defense mechanism in Fusarium graminearum. Biosci Biotechnol Biochem. 1998 May;62(5):1033–1036. doi: 10.1271/bbb.62.1033. [DOI] [PubMed] [Google Scholar]
  7. Kimura Makoto, Anzai Hiroyuki, Yamaguchi Isamu. Microbial toxins in plant-pathogen interactions: Biosynthesis, resistance mechanisms, and significance. J Gen Appl Microbiol. 2001 Aug;47(4):149–160. doi: 10.2323/jgam.47.149. [DOI] [PubMed] [Google Scholar]
  8. Krem M. M., Di Cera E. Molecular markers of serine protease evolution. EMBO J. 2001 Jun 15;20(12):3036–3045. doi: 10.1093/emboj/20.12.3036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kuczuk M. H., Benson P. M., Heath H., Hayes A. W. Evaluation of the mutagenic potential of mycotoxins using Salmonella typhimurium and Saccharomyces cerevisiae. Mutat Res. 1978 Feb;53(1):11–20. doi: 10.1016/0165-1161(78)90375-8. [DOI] [PubMed] [Google Scholar]
  10. Makela S, Davis VL, Tally WC, Korkman J, Salo L, Vihko R, Santti R, Korach KS. Dietary Estrogens Act through Estrogen Receptor-Mediated Processes and Show No Antiestrogenicity in Cultured Breast Cancer Cells. Environ Health Perspect. 1994 Jun;102(6-7):572–578. doi: 10.1289/ehp.94102572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Miksicek R. J. Interaction of naturally occurring nonsteroidal estrogens with expressed recombinant human estrogen receptor. J Steroid Biochem Mol Biol. 1994 Jun;49(2-3):153–160. doi: 10.1016/0960-0760(94)90005-1. [DOI] [PubMed] [Google Scholar]
  12. Nierman W. C., Feldblyum T. V., Laub M. T., Paulsen I. T., Nelson K. E., Eisen J. A., Heidelberg J. F., Alley M. R., Ohta N., Maddock J. R. Complete genome sequence of Caulobacter crescentus. Proc Natl Acad Sci U S A. 2001 Mar 20;98(7):4136–4141. doi: 10.1073/pnas.061029298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Okazaki K., Okazaki N., Kume K., Jinno S., Tanaka K., Okayama H. High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe. Nucleic Acids Res. 1990 Nov 25;18(22):6485–6489. doi: 10.1093/nar/18.22.6485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ollis D. L., Cheah E., Cygler M., Dijkstra B., Frolow F., Franken S. M., Harel M., Remington S. J., Silman I., Schrag J. The alpha/beta hydrolase fold. Protein Eng. 1992 Apr;5(3):197–211. doi: 10.1093/protein/5.3.197. [DOI] [PubMed] [Google Scholar]
  15. Pfohl-Leszkowicz A., Chekir-Ghedira L., Bacha H. Genotoxicity of zearalenone, an estrogenic mycotoxin: DNA adduct formation in female mouse tissues. Carcinogenesis. 1995 Oct;16(10):2315–2320. doi: 10.1093/carcin/16.10.2315. [DOI] [PubMed] [Google Scholar]
  16. Ueno Y., Kubota K. DNA-attacking ability of carcinogenic mycotoxins in recombination-deficient mutant cells of Bacillus subtilis. Cancer Res. 1976 Feb;36(2 Pt 1):445–451. [PubMed] [Google Scholar]
  17. el-Sharkawy S., Abul-Hajj Y. J. Microbial cleavage of zearalenone. Xenobiotica. 1988 Apr;18(4):365–371. doi: 10.3109/00498258809041672. [DOI] [PubMed] [Google Scholar]

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