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Food Microbiology and Biotechnology(FMB)  |  Food Biotechnology

Biodegradation of Ochratoxin A by Aspergillus tubingensis Isolated from Meju

Sung Min Cho 1, Seong Eun Jeong 1, Kyu Ri Lee 1, Hemanth P. K. Sudhani 2, Myunghee Kim 2, Sung-Yong Hong 1 and Soo Hyun Chung 1*

1Department of Integrated Biomedical and Life Science, Korea University, Seoul 02841, Republic of Korea, 2Department of Food Science and Technology, Yeungnam University, Gyeongsan 38541, Republic of Korea

Received: June 9, 2016; Accepted: August 2, 2016

J. Microbiol. Biotechnol. 2016; 26(10): 1687-1695

Published October 28, 2016 https://doi.org/10.4014/jmb.1606.06016

Copyright © The Korean Society for Microbiology and Biotechnology.

Abstract

Ochratoxin A (OTA), a mycotoxin, contaminates agricultural products and poses a serious threat to public health worldwide. Microbiological methods are known to be a promising approach for OTA biodegradation because physical and chemical methods have practical limitations. In the present study, a total of 130 fungal isolates obtained from 65 traditional Korean meju (a fermented starter for fermentation of soybeans) samples were examined for OTA-biodegradation activity using thin-layer chromatography. Two fungal isolates were selected for OTA-biodegradation activity and were identified as Aspergillus tubingensis M036 and M074 through sequence analysis of the beta-tubulin gene. After culturing both A. tubingensis isolates in Soytone-Czapek medium containing OTA (40 ng/ml), OTAbiodegradation activity was analyzed using high-performance liquid chromatography (HPLC). Both A. tubingensis strains degraded OTA by more than 95.0% after 14 days, and the HPLC analysis showed that the OTA biodegradation by the A. tubingensis strains led to the production of ochratoxin α, which is much less toxic than OTA. Moreover, crude enzymes from the cultures of A. tubingensis M036 and M074 led to OTA biodegradation of 97.5% and 91.3% at pH 5, and 80.3% and 75.3% at pH 7, respectively, in a buffer solution containing OTA (40 ng/ml) after 24 h. In addition, the OTA-biodegrading fungi did not exhibit OTA production activity. Our data suggest that A. tubingensis isolates and their enzymes have the potential for practical application to reduce levels of OTA in food and feed.

Keywords

Ochratoxin A, Biodegradation, Aspergilllus tubingensis, Meju

References

  1. Abrunhosa L, Serra R, Venâncio A. 2002. Biodegradation of ochratoxin A by fungi isolated from grapes. J. Agric. Food Chem. 50: 7493-7496.
    Pubmed CrossRef
  2. Abrunhosa L, Venancio A. 2007. Isolation and purification of an enzyme hydrolyzing ochratoxin A from Aspergillus niger. Biotechnol. Lett. 29: 1909-1914.
    Pubmed CrossRef
  3. Alexander J, Autrup H, Bard D, Benford D, Carere A, Costa L, et al. 2006. Opinion of the scientific panel on contaminants in the food chain on a request from the commission related to ochratoxin A in food. EFSA J. 365: 1-56.
  4. Bejaoui H, Mathieu F, Taillandier P, Lebrihi A. 2006. Biodegradation of ochratoxin A by Aspergillus section Nigri species isolated from French grapes: a potential means of ochratoxin A decontamination in grape juices and musts. FEMS Microbiol. Lett. 255: 203-208.
    Pubmed CrossRef
  5. Chang X, Wu Z, Wu S, Dai Y, Sun C. 2015. Degradation of ochratoxin A by Bacillus amyloliquefaciens ASAG1. Food Addit. Contam. Part A Chem. Anal. Control Expo. Risk Assess. 32: 564-571.
    Pubmed CrossRef
  6. Dachoupakan C, Ratomahenina R, Martinez V, Guiraud J, Baccou J, Schorr-Galindo S. 2009. Study of the phenotypic and genotypic biodiversity of potentially ochratoxigenic black aspergilli isolated from grapes. Int. J. Food Microbiol. 132: 14-23.
    Pubmed CrossRef
  7. FAO/WHO (Food and Agricultural Organization/World Health Organization). 2001. Ochratoxin A. In: Safety Evaluations of Specific Mycotoxins. Prepared by the Fifty-Sixth Meeting of the Joint FAO/WHO Expert Committee on Food Additives, 6-15 Feb 2001, Geneva.
  8. Ferenczi S, Cserhati M, Krifaton C, Szoboszlay S, Kukolya J, Szoke Z, et al. 2014. A new ochratoxin A biodegradation strategy using Cupriavidus basilensis Or16 strain. PLoS One 9: e109817.
    Pubmed PMC CrossRef
  9. Gil-Serna J, Vázquez C, Sardiñas N, González-Jaén MT, Patiño B. 2011. Revision of ochratoxin A production capacity by the main species of Aspergillus section Circumdati. Aspergillus steynii revealed as the main risk of OTA contamination. Food Control 22: 343-345.
    CrossRef
  10. Glass NL, Donaldson GC. 1995. Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Appl. Environ. Microbiol. 61:1323-1330.
    Pubmed PMC
  11. He C, Fan Y, Liu G, Zhang H. 2008. Isolation and identification of a strain of Aspergillus tubingensis with deoxynivalenol biotransformation capability. Int. J. Mol. Sci. 9: 2366-2375.
    Pubmed PMC CrossRef
  12. Huwig A, Freimund S, Kappeli O, Dutler H. 2001. Mycotoxin detoxication of animal feed by different adsorbents. Toxicol. Lett. 122: 179-188.
    CrossRef
  13. IARC Working Group on the Evaluation of Carcinogenic Risks to Humans. 2002. Some traditional herbal medicines, some mycotoxins, naphthalene and styrene. IARC Monogr. Eval. Carcinog. Risks Hum. 82: 1-556.
    Pubmed PMC
  14. Jung YJ, Lee HK, Hong SB. 2012. Isolation and identification of fungi from a meju contaminated with aflatoxins. J. Microbiol. Biotechnol. 22: 1740-1748.
    Pubmed CrossRef
  15. McKenzie K, Sarr A, Mayura K, Bailey R, Miller D, Rogers T, et al. 1997. Oxidative degradation and detoxification of mycotoxins using a novel source of ozone. Food Chem. Toxicol. 35: 807-820.
    CrossRef
  16. Medina A, Mateo R, Lopez-Ocana L, Valle-Algarra FM, Jimenez M. 2005. Study of Spanish grape mycobiota and ochratoxin A production by isolates of Aspergillus tubingensis and other members of Aspergillus section Nigri. Appl. Environ. Microbiol. 71: 4696-4702.
    Pubmed PMC CrossRef
  17. Rodriguez H, Reveron I, Doria F, Costantini A, De Las Rivas B, Munoz R, Garcia-Moruno E. 2011. Degradation of ochratoxin A by Brevibacterium species. J. Agric. Food Chem. 59: 10755-10760.
    Pubmed CrossRef
  18. Samson RA, Visagie CM, Houbraken J, Hong S-B, Hubka V, Klaassen CH, et al. 2014. Phylogeny, identification and nomenclature of the genus Aspergillus. Stud. Mycol. 78: 141-173.
    Pubmed PMC CrossRef
  19. Sartori D, Furlaneto MC, Martins MK, de Paula MRF, Pizzirani-Kleiner AA, Taniwaki MH, Fungaro MHP. 2006. PCR method for the detection of potential ochratoxinproducing Aspergillus species in coffee beans. Res. Microbiol. 157: 350-354.
    Pubmed CrossRef
  20. Schatzmayr G, Heidler D, Fuchs E, Binder EM, Loibner AP, Braun R. 2002. Evidence of ochratoxin A-detoxification activity of rumen fluid, intestinal fluid and soil samples as well as isolation of relevant microorganisms from these environments. Mycotoxin Res. 18 Suppl 2: 183-187.
    Pubmed CrossRef
  21. Schatzmayr G, Zehner F, Täubel M, Schatzmayr D, Klimitsch A, Loibner AP, Binder EM. 2006. Microbiologicals for deactivating mycotoxins. Mol. Nutr. Food Res. 50: 543-551.
    Pubmed CrossRef
  22. Schuster E, Dunn-Coleman N, Frisvad J, Van Dijck P. 2002. On the safety of Aspergillus niger – a review. Appl. Microbiol. Biotechnol. 59: 426-435.
    Pubmed CrossRef
  23. Shi L, Liang Z, Li J, Hao J, Xu Y, Huang K, et al. 2014. Ochratoxin A biocontrol and biodegradation by Bacillus subtilis CW 14. J. Sci. Food Agric. 94: 1879-1885.
    Pubmed CrossRef
  24. Skrinjar M, Rasic JL, Stojicic V. 1996. Lowering of ochratoxin A level in milk by yoghurt bacteria and bifidobacteria. Folia Microbiol. (Praha) 41: 26-28.
    CrossRef
  25. Stander MA, Steyn PS, van der Westhuizen FH, Payne BE. 2001. A kinetic study into the hydrolysis of the ochratoxins and analogues by carboxypeptidase A. Chem. Res. Toxicol. 14: 302-304.
    Pubmed CrossRef
  26. Storari M, Dennert FG, Bigler L, Gessler C, Broggini GA. 2012. Isolation of mycotoxins producing black aspergilli in herbal teas available on the Swiss market. Food Control 26:157-161.
    CrossRef
  27. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 28: 27312739.
    Pubmed PMC CrossRef
  28. Varga J, Péteri Z, Tábori K, Téren J, Vágvölgyi C. 2005. Degradation of ochratoxin A and other mycotoxins by Rhizopus isolates. Int. J. Food Microbiol. 99: 321-328.
    Pubmed CrossRef
  29. Wang Y, Wang L, Liu F, Wang Q, Selvaraj JN, Xing F, et al. 2016. Ochratoxin A producing fungi, biosynthetic pathway and regulatory mechanisms. Toxins (Basel) 8: E83.
    Pubmed PMC CrossRef
  30. Xiao H, Marquardt RR, Abramson D, Frohlich AA. 1996. Metabolites of ochratoxins in rat urine and in a culture of Aspergillus ochraceus. Appl. Environ. Microbiol. 62: 648-655.
    Pubmed PMC

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Article

Research article

J. Microbiol. Biotechnol. 2016; 26(10): 1687-1695

Published online October 28, 2016 https://doi.org/10.4014/jmb.1606.06016

Copyright © The Korean Society for Microbiology and Biotechnology.

Biodegradation of Ochratoxin A by Aspergillus tubingensis Isolated from Meju

Sung Min Cho 1, Seong Eun Jeong 1, Kyu Ri Lee 1, Hemanth P. K. Sudhani 2, Myunghee Kim 2, Sung-Yong Hong 1 and Soo Hyun Chung 1*

1Department of Integrated Biomedical and Life Science, Korea University, Seoul 02841, Republic of Korea, 2Department of Food Science and Technology, Yeungnam University, Gyeongsan 38541, Republic of Korea

Received: June 9, 2016; Accepted: August 2, 2016

Abstract

Ochratoxin A (OTA), a mycotoxin, contaminates agricultural products and poses a serious
threat to public health worldwide. Microbiological methods are known to be a promising
approach for OTA biodegradation because physical and chemical methods have practical
limitations. In the present study, a total of 130 fungal isolates obtained from 65 traditional
Korean meju (a fermented starter for fermentation of soybeans) samples were examined for
OTA-biodegradation activity using thin-layer chromatography. Two fungal isolates were
selected for OTA-biodegradation activity and were identified as Aspergillus tubingensis M036
and M074 through sequence analysis of the beta-tubulin gene. After culturing both
A. tubingensis isolates in Soytone-Czapek medium containing OTA (40 ng/ml), OTAbiodegradation
activity was analyzed using high-performance liquid chromatography
(HPLC). Both A. tubingensis strains degraded OTA by more than 95.0% after 14 days, and the
HPLC analysis showed that the OTA biodegradation by the A. tubingensis strains led to the
production of ochratoxin α, which is much less toxic than OTA. Moreover, crude enzymes
from the cultures of A. tubingensis M036 and M074 led to OTA biodegradation of 97.5% and
91.3% at pH 5, and 80.3% and 75.3% at pH 7, respectively, in a buffer solution containing OTA
(40 ng/ml) after 24 h. In addition, the OTA-biodegrading fungi did not exhibit OTA
production activity. Our data suggest that A. tubingensis isolates and their enzymes have the
potential for practical application to reduce levels of OTA in food and feed.

Keywords: Ochratoxin A, Biodegradation, Aspergilllus tubingensis, Meju

References

  1. Abrunhosa L, Serra R, Venâncio A. 2002. Biodegradation of ochratoxin A by fungi isolated from grapes. J. Agric. Food Chem. 50: 7493-7496.
    Pubmed CrossRef
  2. Abrunhosa L, Venancio A. 2007. Isolation and purification of an enzyme hydrolyzing ochratoxin A from Aspergillus niger. Biotechnol. Lett. 29: 1909-1914.
    Pubmed CrossRef
  3. Alexander J, Autrup H, Bard D, Benford D, Carere A, Costa L, et al. 2006. Opinion of the scientific panel on contaminants in the food chain on a request from the commission related to ochratoxin A in food. EFSA J. 365: 1-56.
  4. Bejaoui H, Mathieu F, Taillandier P, Lebrihi A. 2006. Biodegradation of ochratoxin A by Aspergillus section Nigri species isolated from French grapes: a potential means of ochratoxin A decontamination in grape juices and musts. FEMS Microbiol. Lett. 255: 203-208.
    Pubmed CrossRef
  5. Chang X, Wu Z, Wu S, Dai Y, Sun C. 2015. Degradation of ochratoxin A by Bacillus amyloliquefaciens ASAG1. Food Addit. Contam. Part A Chem. Anal. Control Expo. Risk Assess. 32: 564-571.
    Pubmed CrossRef
  6. Dachoupakan C, Ratomahenina R, Martinez V, Guiraud J, Baccou J, Schorr-Galindo S. 2009. Study of the phenotypic and genotypic biodiversity of potentially ochratoxigenic black aspergilli isolated from grapes. Int. J. Food Microbiol. 132: 14-23.
    Pubmed CrossRef
  7. FAO/WHO (Food and Agricultural Organization/World Health Organization). 2001. Ochratoxin A. In: Safety Evaluations of Specific Mycotoxins. Prepared by the Fifty-Sixth Meeting of the Joint FAO/WHO Expert Committee on Food Additives, 6-15 Feb 2001, Geneva.
  8. Ferenczi S, Cserhati M, Krifaton C, Szoboszlay S, Kukolya J, Szoke Z, et al. 2014. A new ochratoxin A biodegradation strategy using Cupriavidus basilensis Or16 strain. PLoS One 9: e109817.
    Pubmed KoreaMed CrossRef
  9. Gil-Serna J, Vázquez C, Sardiñas N, González-Jaén MT, Patiño B. 2011. Revision of ochratoxin A production capacity by the main species of Aspergillus section Circumdati. Aspergillus steynii revealed as the main risk of OTA contamination. Food Control 22: 343-345.
    CrossRef
  10. Glass NL, Donaldson GC. 1995. Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Appl. Environ. Microbiol. 61:1323-1330.
    Pubmed KoreaMed
  11. He C, Fan Y, Liu G, Zhang H. 2008. Isolation and identification of a strain of Aspergillus tubingensis with deoxynivalenol biotransformation capability. Int. J. Mol. Sci. 9: 2366-2375.
    Pubmed KoreaMed CrossRef
  12. Huwig A, Freimund S, Kappeli O, Dutler H. 2001. Mycotoxin detoxication of animal feed by different adsorbents. Toxicol. Lett. 122: 179-188.
    CrossRef
  13. IARC Working Group on the Evaluation of Carcinogenic Risks to Humans. 2002. Some traditional herbal medicines, some mycotoxins, naphthalene and styrene. IARC Monogr. Eval. Carcinog. Risks Hum. 82: 1-556.
    Pubmed KoreaMed
  14. Jung YJ, Lee HK, Hong SB. 2012. Isolation and identification of fungi from a meju contaminated with aflatoxins. J. Microbiol. Biotechnol. 22: 1740-1748.
    Pubmed CrossRef
  15. McKenzie K, Sarr A, Mayura K, Bailey R, Miller D, Rogers T, et al. 1997. Oxidative degradation and detoxification of mycotoxins using a novel source of ozone. Food Chem. Toxicol. 35: 807-820.
    CrossRef
  16. Medina A, Mateo R, Lopez-Ocana L, Valle-Algarra FM, Jimenez M. 2005. Study of Spanish grape mycobiota and ochratoxin A production by isolates of Aspergillus tubingensis and other members of Aspergillus section Nigri. Appl. Environ. Microbiol. 71: 4696-4702.
    Pubmed KoreaMed CrossRef
  17. Rodriguez H, Reveron I, Doria F, Costantini A, De Las Rivas B, Munoz R, Garcia-Moruno E. 2011. Degradation of ochratoxin A by Brevibacterium species. J. Agric. Food Chem. 59: 10755-10760.
    Pubmed CrossRef
  18. Samson RA, Visagie CM, Houbraken J, Hong S-B, Hubka V, Klaassen CH, et al. 2014. Phylogeny, identification and nomenclature of the genus Aspergillus. Stud. Mycol. 78: 141-173.
    Pubmed KoreaMed CrossRef
  19. Sartori D, Furlaneto MC, Martins MK, de Paula MRF, Pizzirani-Kleiner AA, Taniwaki MH, Fungaro MHP. 2006. PCR method for the detection of potential ochratoxinproducing Aspergillus species in coffee beans. Res. Microbiol. 157: 350-354.
    Pubmed CrossRef
  20. Schatzmayr G, Heidler D, Fuchs E, Binder EM, Loibner AP, Braun R. 2002. Evidence of ochratoxin A-detoxification activity of rumen fluid, intestinal fluid and soil samples as well as isolation of relevant microorganisms from these environments. Mycotoxin Res. 18 Suppl 2: 183-187.
    Pubmed CrossRef
  21. Schatzmayr G, Zehner F, Täubel M, Schatzmayr D, Klimitsch A, Loibner AP, Binder EM. 2006. Microbiologicals for deactivating mycotoxins. Mol. Nutr. Food Res. 50: 543-551.
    Pubmed CrossRef
  22. Schuster E, Dunn-Coleman N, Frisvad J, Van Dijck P. 2002. On the safety of Aspergillus niger – a review. Appl. Microbiol. Biotechnol. 59: 426-435.
    Pubmed CrossRef
  23. Shi L, Liang Z, Li J, Hao J, Xu Y, Huang K, et al. 2014. Ochratoxin A biocontrol and biodegradation by Bacillus subtilis CW 14. J. Sci. Food Agric. 94: 1879-1885.
    Pubmed CrossRef
  24. Skrinjar M, Rasic JL, Stojicic V. 1996. Lowering of ochratoxin A level in milk by yoghurt bacteria and bifidobacteria. Folia Microbiol. (Praha) 41: 26-28.
    CrossRef
  25. Stander MA, Steyn PS, van der Westhuizen FH, Payne BE. 2001. A kinetic study into the hydrolysis of the ochratoxins and analogues by carboxypeptidase A. Chem. Res. Toxicol. 14: 302-304.
    Pubmed CrossRef
  26. Storari M, Dennert FG, Bigler L, Gessler C, Broggini GA. 2012. Isolation of mycotoxins producing black aspergilli in herbal teas available on the Swiss market. Food Control 26:157-161.
    CrossRef
  27. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 28: 27312739.
    Pubmed KoreaMed CrossRef
  28. Varga J, Péteri Z, Tábori K, Téren J, Vágvölgyi C. 2005. Degradation of ochratoxin A and other mycotoxins by Rhizopus isolates. Int. J. Food Microbiol. 99: 321-328.
    Pubmed CrossRef
  29. Wang Y, Wang L, Liu F, Wang Q, Selvaraj JN, Xing F, et al. 2016. Ochratoxin A producing fungi, biosynthetic pathway and regulatory mechanisms. Toxins (Basel) 8: E83.
    Pubmed KoreaMed CrossRef
  30. Xiao H, Marquardt RR, Abramson D, Frohlich AA. 1996. Metabolites of ochratoxins in rat urine and in a culture of Aspergillus ochraceus. Appl. Environ. Microbiol. 62: 648-655.
    Pubmed KoreaMed