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Protein-mediated degradation of aflatoxin B1 by Pseudomonas putida

  • Environmental Microbiology - Research Paper
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Abstract

Degradation or the removal of aflatoxin B1 from agriculture commodities is very important because of its acute toxicity and economic loss due to rejection of about 25% contaminated agri produce. The present study aimed at using Pseudomonas putida for the aflatoxin B1 (AFB1) degradation and to understand the mechanism involved. AFB1 degradation was studied with P. putida culture, culture supernatant, cell lysate, cell lysate in the presence of protease inhibitor, and heat-inactivated cell lysate. The remaining AFB1 was qualitatively and quantitatively measured by thin-layer chromatography and HPLC with a UV detector. P. putida culture and culture supernatant showed 80% reduction in AFB1 within 24 h of incubation. Cell lysate and the lysate in the presence of protease inhibitor showed the same reduction in 6 and 4 h respectively. The protease-inhibited lysate showed greater thermostability, broad pH range, and tolerance to some of the solvents and detergents in terms of aflatoxin B1 degrading activity. The heat-inactivated lysate showed only 20% reduction in 24 h of incubation indicating loss of activity on heating. As cell-free supernatant and cell lysate are capable of reducing AFB1 effectively, actively growing cells are not necessary for degradation. The active principle for degradation might be proteinaceous; therefore, heat-inactivated lysate is ineffective for reducing the AFB1. These results showed that degradation of aflatoxin B1 by P. putida might be an enzymatic process and could be used in a broad range of conditions.

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References

  1. Cardwell KF, Desjardins A, Henry SH, Munkvold G (2001) Mycotoxins: the cost of achieving food security and food quality. APSnet Featur Artic. https://doi.org/10.1094/APSnetFeature-2001-0901

  2. Vardon P, McLaughlin C, Nardinelli C (2003) Potential economic costs of mycotoxins in the United States. In: Mycotoxins: risks in plant, animal, and human systems. CAST (Task Force Report) 139:136–142

    Google Scholar 

  3. Robens J, Cardwell K (2003) The costs of mycotoxin management to the USA: management of aflatoxins in the United States. J Toxicol Toxin Rev 22:139–152. https://doi.org/10.1081/TXR-120024089

    Article  Google Scholar 

  4. Mitchell NJ, Bowers E, Hurburgh C, Wu F (2016) Potential economic losses to the US corn industry from aflatoxin contamination. Food Addit Contam Part A 33:540–550. https://doi.org/10.1080/19440049.2016.1138545

    Article  CAS  Google Scholar 

  5. Williams JH, Phillips TD, Jolly PE, Stiles JK, Jolly CM, Aggarwal D (2004) Human aflatoxicosis in developing countries: a review of toxicology, exposure, potential health consequences, and interventions. Am J of Clin Nutr 80:1106–1122. https://doi.org/10.1093/ajcn/80.5.1106

    Article  CAS  Google Scholar 

  6. International Agency for Research on Cancer (IARC) (2002) Some traditional herbal medicines, some mycotoxins, naphthalene and styrene: monograph on the evaluation of carcinogenic risks to humans. IARC, Lyon, p 82

    Google Scholar 

  7. Sarajeeamwa U, Sen AC, Cohen MD, Wei CI (1990) Detoxification of aflatoxins in foods and feeds by physical and chemical methods. J Food Prot 53:489–501

    Article  Google Scholar 

  8. Rustom IYS (1997) Aflatoxin in food and feed: occurrence, legislation and inactivation by physical methods. Food Chem 59:57–67. https://doi.org/10.1016/S0308-8146(96)00096-9

    Article  CAS  Google Scholar 

  9. Doyle MP, Applebaum RS, Brackett RE, Marth EH (1982) Physical, chemical and biological degradation of mycotoxins in foods and agricultural commodities. J Food Prot 45:964–971. https://doi.org/10.4315/0362-028X-45.10.964

    Article  CAS  PubMed  Google Scholar 

  10. Haskard CA, El-Nezami HS, Kankaanpää PE, Salminen S, Ahokas JT (2001) Surface binding of aflatoxin B1 by lactic acid bacteria. Appl Environ Microbiol 67:3086–3091. https://doi.org/10.1128/AEM.67.7.3086-3091.2001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Peltonen K, El-Nezami H, Haskard C, Ahokas J, Salminen S (2001) Aflatoxin B1 binding by dairy strains of lactic acid bacteria and Bifidobacteria. J Dairy Sci 84:2152–2156. https://doi.org/10.3168/jds.S0022-0302(01)74660-7

    Article  CAS  PubMed  Google Scholar 

  12. El-Nezami HS, Chrevatidis A, Auriola S, Salminen S, Mykkänen H (2002) Removal of common Fusarium toxins in vitro by strains of Lactobacillus and Propionibacterium. Food Addit Contam 19:680–686. https://doi.org/10.1080/02652030210134236

    Article  CAS  PubMed  Google Scholar 

  13. Fuchs S, Sontag G, Stidl R, Ehrlich V, Kundi M, Knasmüller S (2008) Detoxification of patulin and ochratoxin A, two abundant mycotoxins, by lactic acid bacteria. Food Chem Toxicol 46:1398–1407. https://doi.org/10.1016/j.fct.2007.10.008

    Article  CAS  PubMed  Google Scholar 

  14. Shu X, Wang Y, Zhou Q, Li M, Hu H, MaY WL (2018) Biological degradation of aflatoxin B1 by cell-free extracts of Bacillus velezensis DY3108 with broad pH stability and excellent thermostability. Toxins 10:330

    Article  Google Scholar 

  15. Ahlberg SH, Joutsjoki V, Korhonen HJ (2015) Potential of lactic acid bacteria in aflatoxin risk mitigation. Int J Food Microbiol 207:87–102. https://doi.org/10.1016/j.ijfoodmicro.2015.04.042

    Article  CAS  PubMed  Google Scholar 

  16. Liew WPP, Adilah ZN, Than LTL, Redzwan SM (2018) The binding efficiency and interaction of Lactobacillus casei Shirota toward aflatoxin B1. Front Microbiol 9:1503. https://doi.org/10.3389/fmicb.2018.01503

    Article  PubMed  PubMed Central  Google Scholar 

  17. Huang L, Duan C, Zhao Y, Gao L, Niu C, Xu J Li S (2017) Reduction of aflatoxin B1 toxicity by Lactobacillus plantarum C88: a potential probiotic strain isolated from Chinese traditional fermented food “tofu”. PLoS One 12:e0170109. https://doi.org/10.1371/journal.pone.0170109

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Alberts JF, Gelderblom WCA, Botha A, van Zyl WH (2009) Degradation of aflatoxin B1 by fungal laccase enzymes. Int J Food Microbiol 135:47–52. https://doi.org/10.1016/j.ijfoodmicro.2009.07.022

    Article  CAS  PubMed  Google Scholar 

  19. Teniola O, Addo P, Brost I, Farber P, Jany K, Alberts J, Vanzyl W, Steyn P, Holzapfel W (2005) Degradation of aflatoxin B1 by cell-free extracts of Rhodococcus erythropolis and Mycobacterium fluoranthenivorans sp. nov. DSM44556T. Int J Food Microbiol 105:111–117. https://doi.org/10.1016/j.ijfoodmicro.2005.05.004

    Article  CAS  PubMed  Google Scholar 

  20. Alberts J, Engelbrecht Y, Steyn P, Holzapfel W, Vanzyl W (2006) Biological degradation of aflatoxin B1 by Rhodococcus erythropolis cultures. Int J Food Microbiol 109:121–126. https://doi.org/10.1016/j.ijfoodmicro.2006.01.019

    Article  CAS  PubMed  Google Scholar 

  21. Guan S, Ji C, Zhou T, Li J, Ma Q, Niu T (2008) Aflatoxin B1 degradation by Stenotrophomonas maltophilia and other microbes selected using coumarin medium. Int J Mol Sci 9:1489–1503. https://doi.org/10.3390/ijms9081489

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Otenio MH, Lopes Da Silva MT, Marques MLO, Roseiro JC, Bidoia ED (2005) Benzene, toluene and xylene biodegradation by Pseudomonas putida CCMI 852. Brazilian J Microbiol 36:258–261. https://doi.org/10.1590/S1517-83822005000300010

    Article  CAS  Google Scholar 

  23. Dunn NW, Gunsalus IC (1973) Transmissible plasmid coding early enzymes of naphthalene oxidation in Pseudomonas putida. J Bacteriol 114:974–979

    Article  CAS  Google Scholar 

  24. Williams PA, Murray K (1974) Metabolism of benzoate and the methylbenzoates by Pseudomonas putida (arvilla) mt-2: evidence for the existence of a TOL plasmid. J Bacteriol 120:416–423

    Article  CAS  Google Scholar 

  25. Franklin FC, Bagdasarian M, Bagdasarian MM, Timmis KN (1981) Molecular and functional analysis of the TOL plasmid pWWO from Pseudomonas putida and cloning of genes for the entire regulated aromatic ring meta cleavage pathway. Proc Natl Acad Sci U S A 78:7458–7462

    Article  CAS  Google Scholar 

  26. Verheecke C, Liboz T, Mathieu F (2016) Microbial degradation of aflatoxin B1: current status and future advances. Int J Food Microbiol 237:1–9. https://doi.org/10.1016/j.ijfoodmicro.2016.07.028

    Article  CAS  PubMed  Google Scholar 

  27. Taylor MC, Jackson CJ, Tattersall DB, French N, Peat TS, Newman J, Briggs LJ, Lapalikar GV, Campbell PM, Scott C, Russell RJ, Oakeshott JG (2010) Identification and characterization of two families of F420H2-dependent reductases from Mycobacteria that catalyse aflatoxin degradation. Mol Microbiol 78:561–575. https://doi.org/10.1111/j.1365-2958.2010.07356

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Yehia RS (2014) Aflatoxin detoxification by manganese peroxidase purified from Pleurotus ostreatus. Brazilian J Microbiol 45:127–133. https://doi.org/10.1590/S1517-83822014005000026

    Article  Google Scholar 

  29. Samuel MS, Sivaramakrishna A, Mehta A (2014) Degradation and detoxification of aflatoxin B1 by Pseudomonas putida. Int Biodeterior Biodegradation 86:202–209. https://doi.org/10.1016/j.ibiod.2013.08.026

    Article  CAS  Google Scholar 

  30. Aiko V, Mehta A (2013) Control of Aspergillus flavus and aflatoxin production with spices in culture medium and rice. World Mycotoxin J 6:43–50. https://doi.org/10.3920/WMJ2012.1437

    Article  CAS  Google Scholar 

  31. Kulkarni M, Chaudhari A (2006) Efficient Pseudomonas putida for degradation of p-nitrophenol. Indian J Biotechnol 5:411–415

    CAS  Google Scholar 

  32. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3

    Article  CAS  Google Scholar 

  33. Sangare L, Zhao Y, Folly YME, Chang J, Li J, Selvaraj JN, Xing F, Zhou L, Wang Y, Liu Y (2014) Aflatoxin B1 degradation by a Pseudomonas strain. Toxins 6:3028–3040

    Article  CAS  Google Scholar 

  34. Smiley RD, Draughon FA (2000) Preliminary evidence that degradation of aflatoxin B1 by Flavobacterium aurantiacum is enzymatic. J Food Prot 63:415–418

    Article  CAS  Google Scholar 

  35. Adebo OA, Njobeh PB, Gbashi S, Nwinyi OC, Mavumengwana V (2017) Review on microbial degradation of aflatoxins. Crit Rev Food Sci Nutr 57:3208–3217. https://doi.org/10.1080/10408398.2015.1106440

    Article  CAS  PubMed  Google Scholar 

  36. Guan S, Zhao L, Ma Q, Zhou T, Wang N, Hu X, Ji C (2010) In vitro efficacy of Myxococcus fulvus ANSM068 to biotransform aflatoxin B1. Int J Mol Sci 11:4063–4079. https://doi.org/10.3390/ijms11104063

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

The authors are grateful to the management of Vellore Institute of Technology, Vellore, Tamilnadu, India, for providing the facilities and fundings for the presented work. Ms. Jyoti Singh is also thankful to the Vellore Institute of Technology for providing her an institutional research fellowship.

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  1. Department of Integrative Biology, School of Biosciences and Technology, Vellore Institute of Technology, Vellore, Tamil Nadu, 632014, India

    Jyoti Singh & Alka Mehta

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Correspondence to Alka Mehta.

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Singh, J., Mehta, A. Protein-mediated degradation of aflatoxin B1 by Pseudomonas putida. Braz J Microbiol 50, 1031–1039 (2019). https://doi.org/10.1007/s42770-019-00134-x

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