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Characterization of Three Extracellular β-Glucosidases Produced by a Fungal Isolate Aspergillus sp. YDJ14 and Their Hydrolyzing Activity for a Flavone Glycoside

  • Oh, Jong Min (Department of Pharmacy, and Research Institute of Life Pharmaceutical Sciences, Sunchon National University) ;
  • Lee, Jae Pil (Department of Pharmacy, and Research Institute of Life Pharmaceutical Sciences, Sunchon National University) ;
  • Baek, Seung Cheol (Department of Pharmacy, and Research Institute of Life Pharmaceutical Sciences, Sunchon National University) ;
  • Jo, Yang Do (Department of Agricultural Chemistry, Sunchon National University) ;
  • Kim, Hoon (Department of Pharmacy, and Research Institute of Life Pharmaceutical Sciences, Sunchon National University)
  • Received : 2018.02.28
  • Accepted : 2018.03.14
  • Published : 2018.05.28

Abstract

A cellulolytic fungus, YDJ14, was isolated from compost and identified as an Aspergillus sp. strain. Three extracellular ${\beta}$-glucosidases, BGL-A1, BGL-A2, and BGL-A3, were separated using ultrafiltration, ammonium sulfate fractionation, and High-Q chromatography. The molecular masses of the three enzymes were estimated to be 100, 45, and 40 kDa, respectively, by SDS-PAGE. The optimum pH and temperature of BGL-A3 were 5.0 and $50^{\circ}C$, respectively, whereas the optimum pH and temperature of BGL-A1 and BGL-A2 were identical (4.0 and $60^{\circ}C$, respectively). The half-life of BGL-A3 at $70^{\circ}C$ (2.8 min) was shorter than that of BGL-A1 and BGL-A2 (12.1 and 8.8 min, respectively). All three enzymes preferred p-nitrophenyl-${\beta}$-$\text\tiny{D}$-glucopyranoside (pNPG) and hardly hydrolyzed cellobiose, suggesting that these enzymes were aryl ${\beta}$-glucosidases. The $K_m$ of BGL-A3 (1.26 mM) for pNPG was much higher than that of BGL-A1 and BGL-A2 (0.25 and 0.27 mM, respectively). These results suggested that BGL-A1 and BGL-A2 were similar in their enzymatic properties, whereas BGL-A3 differed from the two enzymes. When tilianin (a flavone glycoside of acacetin) was reacted with the three enzymes, the inhibitory activity for monoamine oxidase, a target in the treatment of neurological disorders, was similar to that shown by acacetin. We conclude that these enzymes may be useful in the hydrolysis of flavone glycosides to improve their inhibitory activities.

Keywords

References

  1. Ketudat Cairns JR, Esen A. 2010. ${\beta}$-Glucosidases. Cell. Mol. Life Sci. 67: 3389-3405. https://doi.org/10.1007/s00018-010-0399-2
  2. Sorensen A, Lubeck M, Lubeck PS, Ahring BK. 2013. Fungal beta-glucosidases: a bottleneck in industrial use of lignocellulosic materials. Biomolecules 3: 3612-3631.
  3. Patchett ML, Daniel RM, Morgan HW. 1987. Purification and properties of a stable beta-glucosidase from an extremely thermophilic anaerobic bacterium. Biochem. J. 243: 779-787. https://doi.org/10.1042/bj2430779
  4. Ahmed A, Nasim Fu-H, Batool K, Bibi A. 2017. Microbial ${\beta}$-glucosidase: sources, production and applications. J. Appl. Environ. Microbiol. 5: 31-46.
  5. Krisch J, Tako M, Papp T, Vagvolgyi C. 2010. Characteristics and potential use of ${\beta}$-glucosidases from Zygomycetes, pp. 891-896. In Mendez-Vilas A (ed.). Current Research, Technology and Education Topics in Applied Microbiology and Microbial Biotechnology. Formatex Research Center, Badajoz, Spain.
  6. Singh G, Verma AK, Kumar V. 2016. Catalytic properties, functional attributes and industrial applications of ${\beta}$-glucosidases. 3 Biotech 6: 3.
  7. Tokpohozin SE, Fischer S, Sacher B, Becker T. 2016. ${\beta}$-D Glucosidase as "key enzyme" for sorghum cyanogenic glucoside (dhurrin) removal and beer bioflavouring. Food Chem. Toxicol. 97: 217-223. https://doi.org/10.1016/j.fct.2016.09.015
  8. Cao P, Wang L, Wang Y, Zhou N, Chen Y. 2015. Alkali- tolerant ${\beta}$-glucosidase produced by newly isolated Aspergillus fumigatus WL002 from rotten wood. Int. Biodeterior. Biodegradation 105: 276-282. https://doi.org/10.1016/j.ibiod.2015.09.010
  9. Sorensen A, Ahring BK, Lubeck M, Ubhayasekera W, Bruno KS, Culley DE, et al. 2012. Identifying and characterizing the most significant ${\beta}$-glucosidase of the novel species Aspergillus saccharolyticus. Can. J. Microbiol. 58: 1035-1046. https://doi.org/10.1139/w2012-076
  10. Thongpoo P, Srisomasap C, Chokchaichamnankit D, Kitpreechavanich V, Svasti J, Kongsaeree PT. 2014. Purification and characterization of three ${\beta}$-glycosidases exhibiting high glucose tolerance from Aspergillus niger ASKU28. Biosci. Biotechnol. Biochem. 78: 1167-1176. https://doi.org/10.1080/09168451.2014.915727
  11. Asha P, Jose Divya, Bright Singh IS. 2016. Purification and characterisation of processive-type endoglucanase and ${\beta}$-glucosidase from Aspergillus ochraceus MTCC 1810 through saccharification of delignified coir pith to glucose. Bioresour. Technol. 213: 245-248. https://doi.org/10.1016/j.biortech.2016.03.013
  12. Kudo K, Watanabe A, Ujiie S, Shintani T, Gomi K. 2015. Purification and enzymatic characterization of secretory glycoside hydrolase family 3 (GH3) aryl ${\beta}$-glucosidases screened from Aspergillus oryzae genome. J. Biosci. Bioeng. 120: 614-623. https://doi.org/10.1016/j.jbiosc.2015.03.019
  13. Yan FY, Xia W, Zhang XX, Chen S, Nie XZ, Qian LC. 2016. Characterization of ${\beta}$-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones. J. Zhejiang Univ. Sci. B 17: 455-464. https://doi.org/10.1631/jzus.B1500317
  14. Chang KH, Jo MN, Kim KT, Paik HD. 2012. Purification and characterization of a ginsenoside Rb(1)-hydrolyzing ${\beta}$-glucosidase from Aspergillus niger KCCM 11239. Int. J. Mol. Sci. 13: 12140-12152. https://doi.org/10.3390/ijms130912140
  15. Oh JM, Lee JP, Baek SC, Kim SG, Jo YD, Kim J, et al. 2018. Characterization of two extracellular ${\beta}$-glucosidases produced from the cellulolytic fungus Aspergillus sp. YDJ216 and their potential applications for the hydrolysis of flavone glycosides. Int. J. Biol. Macromol. 111: 595-603. https://doi.org/10.1016/j.ijbiomac.2018.01.020
  16. Zang X, Liu M, Wang H, Fan Y, Zhang H, Liu J, et al. 2017. The distribution of active ${\beta}$-glucosidase-producing microbial communities in composting. Can. J. Microbiol. 63: 998-1008. https://doi.org/10.1139/cjm-2017-0368
  17. Yan FY, Xia W, Zhang XX, Chen S, Nie XZ, Qian LC. 2016. Characterization of ${\beta}$-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones. J. Zhejiang Univ. Sci. B 17: 455-464. https://doi.org/10.1631/jzus.B1500317
  18. Park DJ, Lee YS, Choi YL. 2013. Characterization of a cold- active ${\beta}$-glucosidase from Paenibacillus xylanilyticus KJ-03 capable of hydrolyzing isoflavones daidzin and genistin. Protein J. 32: 579-584. https://doi.org/10.1007/s10930-013-9520-3
  19. Yang X, Ma R, Shi P, Huang H, Bai Y, Wang Y, et al. 2014. Molecular characterization of a highly-active thermophilic ${\beta}$-glucosidase from Neosartorya fischeri P1 and its application in the hydrolysis of soybean isoflavone glycosides. PLoS One 9: e106785. https://doi.org/10.1371/journal.pone.0106785
  20. Shin KC, Nam HK, Oh DK. 2013. Hydrolysis of flavanone glycosides by ${\beta}$-glucosidase from Pyrococcus furiosus and its application to the production of flavanone aglycones from citrus extracts. J. Agric. Food Chem. 61: 11532-11540. https://doi.org/10.1021/jf403332e
  21. Lee HW, Ryu HW, Baek SC, Kang MG, Park D, Han HY, et al. 2017. Potent inhibitions of monoamine oxidase A and B by acacetin and its 7-O-(6-O-malonylglucoside) derivative from Agastache rugosa. Int. J. Biol. Macromol. 104: 547-553. https://doi.org/10.1016/j.ijbiomac.2017.06.076
  22. Cho KM, Kwon EJ, Kim SK, Kambiranda DM, Math RK, Lee YH, et al. 2009. Fungal diversity in composting process of pig manure and mushroom cultural waste based on partial sequence of large subunit rRNA. J. Microbiol. Biotechnol. 19: 743-748.
  23. Zhang Z, Schwartz S, Wagner L, Miller W. 2000. A greedy algorithm for aligning DNA sequences. J. Comput. Biol. 7: 203-214. https://doi.org/10.1089/10665270050081478
  24. Morgulis A, Coulouris G, Raytselis Y, Madden TL, Agarwala R, Schaffer AA. 2008. Database indexing for production MegaBLAST searches. Bioinformatics 24: 1757-1764. https://doi.org/10.1093/bioinformatics/btn322
  25. Jeong YS, Na HB, Kim SK, Kim YH, Kwon EJ, Kim J, et al. 2012. Characterization of Xyn10J, a novel family 10 xylanase from a compost metagenomic library. Appl. Biochem. Biotechnol. 166: 1328-1339. https://doi.org/10.1007/s12010-011-9520-8
  26. Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254. https://doi.org/10.1016/0003-2697(76)90527-3
  27. Laemmli UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685. https://doi.org/10.1038/227680a0
  28. Phuong ND, Jeong YS, Selvaraj T, Kim SK, Kim YH, Jung KH, et al. 2012. Production of XynX, a large multimodular protein of Thermoanaerobacterium sp., by protease-deficient Bacillus subtilis strains [corrected], Appl. Biochem. Biotechnol 168: 375-382. Appl. Biochem. Biotechnol. 168: 1349-1350. https://doi.org/10.1007/s12010-012-9900-8
  29. Miller GL. 1959. Use of dinitrosalicylic acid reagent for the determination of reducing sugar. Anal. Chem. 31: 428-436.
  30. Shin ES, Yang MJ, Jung KH, Kwon EJ, Jung JS, Park SK, et al. 2002. Influence of the transposition of the thermostabilizing domain of Clostridium thermocellum xylanase (XynX) on xylan binding and thermostabilization. Appl. Environ. Microbiol. 68: 3496-3501. https://doi.org/10.1128/AEM.68.7.3496-3501.2002
  31. Kwon KS, Kang HG, Hah YC. 1992. Purification and characterization of two extracellular ${\beta}$-glucosidases from Aspergillus nidulans. FEMS Microbiol Lett. 76: 149-153.
  32. Madhu KM, Beena PS, Chandrasekaran M. 2009. Extracellular ${\beta}$-glucosidase production by a marine Aspergillus sydowii BTMFS 55 under solid state fermentation using statistical experimental design. Biotechnol. Bioprocess Eng. 14: 457-466. https://doi.org/10.1007/s12257-008-0116-2
  33. Kalyani D, Lee KM, Tiwari MK, Ramachandran P, Kim H, Kim IW, et al. 2012. Characterization of a recombinant aryl ${\beta}$-glucosidase from Neosartorya fischeri NRRL181. Appl. Microbiol. Biotechnol. 94: 413-423. https://doi.org/10.1007/s00253-011-3631-6
  34. Li X, Zhao J, Shi P, Yang P, Wang Y, Luo H, et al. 2013. Molecular cloning and expression of a novel ${\beta}$-glucosidase gene from Phialophora sp. G5. Appl. Biochem. Biotechnol. 169: 941-949. https://doi.org/10.1007/s12010-012-0048-3
  35. Plant AR, Oliver JE, Patchett ML, Daniel RM, Morgan HW. 1988. Stability and substrate specificity of a ${\beta}$-glucosidase from the thermophilic bacterium Tp8 cloned into Escherichia coli. Arch. Biochem. Biophys. 262: 181-188. https://doi.org/10.1016/0003-9861(88)90180-4
  36. Gong G, Zheng Z, Liu H, Wang L, Diao J, Wang P, et al. 2014. Purification and characterization of a ${\beta}$-glucosidase from Aspergillus niger and its application in the hydrolysis of geniposide to genipin. J. Microbiol. Biotechnol. 24: 788-794. https://doi.org/10.4014/jmb.1401.01053
  37. Pei X, Zhao J, Cai P, Sun W, Ren J, Wu Q, et al. 2016. Heterologous expression of a GH3 ${\beta}$-glucosidase from Neurospora crassa in Pichia pastoris with high purity and its application in the hydrolysis of soybean isoflavone glycosides. Protein Expr. Purif. 119: 75-84. https://doi.org/10.1016/j.pep.2015.11.010
  38. Yan Q, Zhou XW, Zhou WX, Li W, Feng MQ, Zhou P. 2008. Purification and properties of a novel ${\beta}$-glucosidase, hydrolyzing ginsenoside Rb1 to CK, from Paecilomyces Bainier. J. Microbiol. Biotechnol. 18: 1081-1089.
  39. Lin F, Guo X, Lu W. 2015. Efficient biotransformation of ginsenoside Rb1 to Rd by isolated Aspergillus versicolor, excreting ${\beta}$-glucosidase in the spore production phase of solid culture. Antonie Van Leeuwenhoek 108: 1117-1127. https://doi.org/10.1007/s10482-015-0565-5

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