Isolation and identification of an extracellular enzyme from Aspergillus Niger with Deoxynivalenol biotransformation capability
DOI:
https://doi.org/10.9755/ejfa.2017.v29.i10.1295Keywords:
Aspergillus niger, DON biotransformation, enzyme, identification, purificationAbstract
In this study, the purification and characterization of an extracellular enzyme form Aspergillus niger was performed. With an optimized protocol, it was conducted a 42.6-fold purification with a yield of 26.2%. The purified lipase had a monomeric molecular weight of 40.5kDa and an isoelectric point of 6.01, and its maximum enzyme activity could be achieved at 40°C and pH 7.5-9.0. The enzyme could be activated by Ca2+, Mg2+ and Fe2+, while its activity could be inhibited by Zn2+ and Cu2+. Additionally, organic compounds exerted an inhibitory effect on the enzyme activity in a descending order of methanol, ethanol, DMSO, EDTA, acetone. Meanwhile, the specificity analysis of the enzyme indicated a preference to tributyrin and vegetable oils as well as long-chain fatty acid methyl esters (C12-C18). Most importantly, this enzyme could successfully transform deoxynivalenol (DON). Using HPLC analysis，it was detected a biotransformation rate of more than 70%.The liquid chromatography-mass spectrometry (LC-MS) analysis showed that the molecular weight of the transformation product was 18.0 larger than that of DON, indicating that DON could be hydrolyzed by the enzyme. Overall, the proposed method here provides a new avenue for reducing the toxicity of DON, which appears to have a wide application outlook for DON biotransformation.
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References
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Zou, Z.Y., Z.F. He, H.J. Li, P.F. Han, J. Tang, C. Xi, Y. Li, L. Zhang and X. Li. 2012. Development and application of a method for the analysis of two trichothecenes: Deoxynivalenol and T-2 toxin in meat in China by HPLC-MS/MS. Meat Sci.90:613-617.
Alassane, K., P. Imourana and O. Olivier. 2015. Toxicological interactions between the mycotoxins deoxynivalenol, nivalenol and their acetylated derivatives in intestinal epithelial cells. Arch. Toxicol. 89: 1337-1346.
Ali, M.S., C.C. Yun, A.L. Chor, R.N. Rahman, M. Basri, A.B. Salleh and A.L. Chor. 2012. Purification and Characterisation of an F16L Mutant of a thermostable lipase. Protein J. 31: 229-237.
Avantaggiato, G., R. Havenaar and A. Visconti. 2004. Evaluation of the intestinal absorption of deoxynivalenol and nivalenol by an in vitro gastrointestinal model, and the binding efficacy of activated carbon and other adsorbent materials.Food Chem. Toxicol. 42:817-824.
Awad, W.A., K. Ghareeb, J. Bohm and J. Zentek. 2010. Decontamination and detoxification strategies for the Fusarium mycotoxin deoxynivalenol in animal feed and the effectiveness of microbial biodegradation. Food Addit. Contam. 4:510-520.
Bretz, M., M.Beyer, B. Cramer. A. Knecht and H.U. Humpf. 2006.Thermal degradation of the Fusarium mycotoxin deoxynivalenol. J. Agric. Food Chem. 54: 6445-6451.
Castro-Ochoa, L.D., C.R. Gomez, G.V. Alfaro and R.O. Ros. 2005. Screening, purification and characterization of the thermoalkalophilic lipase produced by Bacillus thermoleovoransCCR11.Enzyme Microb. Technol. 37: 648-654.
He, C., Y. Fan, G. Liu and H. Zhang. 2008. Isolation and identification of a strain of Aspergillus tubingensis with deoxynivalenol biotransformation capability. Int. J. Mol. Sci.9: 2366-2375.
Lukaszczyk, J. and K. Jaszcz. 2002. Studies on hydrolytic degradation of epoxy-polyester resins cured with glutaric anhydride. Polym Adv. Technol. 13:871-883.
Jurgens, D., H.Huser and H. Brunner. 1981. Purification and characterization of Staphilococcus aureus lipase.FEMS Microbiol. Lett.12: 195-199.
Kim, J.T., S.G.Kang, J.H.Woo, J.H.Lee, B.C.Jeong and S.J. Kim. 2007. Screening and its potential application of lipolytic activity from a marine environment: characterization of a novel esterase from Yarrowia lipolytica CL180. Appl. Microbiol. Biot.74:820-828.
Kumar, S., K. Kikon and A.Upadhyay. 2005. Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulansBTS-3.Protein Expres. Purif. 41: 38-44.
Laemmli, U.K.1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nat. 227: 680-685.
Mase, T., Y. Matsumiya and T.Akiba.1995. Purification and characterization of a new lipase from Fusariumsp YM-30.Biosci. Biotechnol. Biochem. 59:1771-1772.
Mateo, C., A. Archelas and R. Furstoss.2003. A spectrophotometric assay for measuring and detecting an epoxide hydrolase activity.Anal.Biochem.314: 135-141.
Mhetras, N.C., K.B. Bastawde and D.V. Gokhale. 2009. Purification and characterization of acidic lipase from Aspergillus niger NCIM 1207.Bioresour. Technol. 100:1486-1490.
Palomo, J.M., G.Munoz,G.Fernandez-Lorente,C.Mateo, M.Fuentes, J.M. Guisan and R.Fernandez-Lafuente. 2003.Modulation of Mucor miehei lipase properties via directed immobilization on different hetero-functional epoxy resins: Hydrolytic resolution of (R, S)-2-butyroyl-2-phenylacetic acid. J. Mol. Catal.21: 201-210.
Papadopoulou-Bouraoui, A., T. Vrabcheva, S.Valzacchi, J. Stroka and E.Anklam. 2004. Screening survey of deoxynivalenol in beer from the European market by an enzyme-linked immunosorbent assay. Food Addit. Contam.21: 607-617.
Pignede, G., H. Wang, F. Fudalej, C. Gaillardin, M. Seman and J.M. Nicaud. 2000. Characterization of an extracellular lipase encoded by LIP2 in Yarrowia lipolytica. J. Bacteriol.182: 2802-2810.
Pinton, P. and P. O. Isabelle. 2014. Effect of Deoxynivalenol and Other Type B Trichothecenes on the Intestine: A Review. Toxions.6:1615-1643.
Reetz, M.T., C. Torre, A. Eipper, R. Lohmer, M. Hermes, B. Brunner, A. Maichele, M. Bocola, M. Arand, A. Cronin,Y. Genzel, A. Archelas and R. Furstoss. 2004. Enhancing the enantioselectivity of an epoxide hydrolase by directed evolution.Org. Lett. 6: 177-80.
Rigo, E., J. L. Ninow and S.M. Tsai. 2012. A preliminary characterization of novel extra-cellular lipase from Penicillium crustosum under solid-state fermentation and its potential application for triglycerides hydrolysis. Food Bioprocess Technol. 5: 1592-1600.
Shim, J., S. Takase, Y. Takahashi, Y. Iwai, H. Fujimoto, M.Yamazaki and K. Ochi. 1997. Novel detoxification of the trichothecene mycotoxin deoxynivalenol by a soil bacterium isolated by enrichment culture. Appl. Env. Microbiol. 63:3825-3830.
Taiana, D.S., S.S. Caldas, G.P. Edneiand B.F.Eliana.2015. Exposure to deoxynivalenol, Ht-2 and T-2 toxins by consumption of wheat-based product in southern Brazil. Food Cont.50: 789-793.
Tay, F.R., D.H. Pashley, R.J. Loushine , S. Kuttler , F. García-Godoy , N.M. King and M. Ferrari . 2005. Susceptibility of a polycaprolactone-based root canal filling material to degradation.J.endod.31:593-598.
Tiemann, U., T. Viergutz, L. Jonas and F. Schneider. 2003. Influence of the mycotoxins α- and β-zearalenol and deoxynivalenol on the cell cycle of cultured porcine endometrial cells. Rep. Toxicol. 17: 209-218.
Treichel, H., D.D.Oliveira,M.A.Mazutti,M.D.Luccio,and J.V.Oliveira. 2010.A Review on microbial lipases production. Food Bioprocess Technol.3:182-196.
Ulker, S. and S.A.Karaoglu. 2012.Purification and characterization of an extracellular lipase from Mucor hiemalis f. corticola isolated from soil.J. Biosci. Bioeng.4:385-390.
Wang, H., S. Zhong and H. Ma.2012. Screening and characterization of a novel alkaline lipase from Acinetobacter calcoaceticus 1-7 isolated from Bohai Bay in China for detergent formulation.J. Microbiol.43:148-156.
Young, J.C., T. Zhou, H. Yu, H. Zhu and J.Gong. 2007. Degradation of trichothecene mycotoxins by chicken intestinal microbes. Food Chem. Toxicol. 45:136-143.
Yu, M., S. Qinand T. Tan. 2007. Purification and characterization of the extracellular lipase Lip2 fromYarrowia lipolytica. Process Biochem.42: 384-391.
Zou, Z.Y., Z.F. He, H.J. Li, P.F. Han, J. Tang, C. Xi, Y. Li, L. Zhang and X. Li. 2012. Development and application of a method for the analysis of two trichothecenes: Deoxynivalenol and T-2 toxin in meat in China by HPLC-MS/MS. Meat Sci.90:613-617.
Published
2017-11-04
How to Cite
Yang, S., Y. Wu, J. Yang, R. Yan, Y. Bao, K. Wang, G. Liu, and W. Wang. “Isolation and Identification of an Extracellular Enzyme from Aspergillus Niger With Deoxynivalenol Biotransformation Capability”. Emirates Journal of Food and Agriculture, vol. 29, no. 10, Nov. 2017, pp. 742-50, doi:10.9755/ejfa.2017.v29.i10.1295.
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Regular Articles
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