The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
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Production and Characterization of the Beneficial β-glucuronidase Inhibitor from non pathogenic wild yeast, Candida oleophila WP5-19-1, and its effects on gut microbes

비병원성 야생효모 Candida oleophila WP5-19-1로부터 장내유용세균 유해 효소 β-glucuronidase 저해제의 생산 및 특성

  • Seon-Jeong Park (Department of Biomedicine and Biotechnology, Paichai University) ;
  • Jong-Soo Lee (Department of Biomedicine and Biotechnology, Paichai University)
  • 박선정 (배재대학교 바이오의약학부) ;
  • 이종수 (배재대학교 바이오의약학부)
  • Received : 2022.01.27
  • Accepted : 2022.01.22
  • Published : 2022.03.31
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Abstract

We aimed to produce a potent β-glucuronidase inhibitor from wild yeast that could inactivate toxic substances in the colon. Culture supernatants and cell-free extracts of non-pathogenic wild yeasts were prepared and their β-glucuronidase inhibitory activities were measured. Cell-free extract from Candida oleophila WP5-19-1 showed the highest β-glucuronidase inhibitory activity (49.0%). The β-glucuronidase inhibitor was maximally produced (IC50 value; 8.4 mg) when C. oleophila WP5-19-1 was cultured in potato dextrose medium containing 5% dextrose (initial pH; 6.0) at 30℃ for 24 hours. β-glucuronidase inhibitor of C. oleophila WP5-19-1 was partially purified by trypsin hydrolysis, ultrafiltration (3 kDa), and Sephadex G-50 filtration. The partially purified β-glucuronidase inhibitor was stable from 30℃ to 60℃ and at pH 6.0 9.0, and showed residual inhibitory activity of about 80%.

비병원성 야생효모로부터 대장에서의 독성물질 활성화에 관여하여 대장암을 유발시키는 효소 중의 하나인 β-glucuronidase를 저해하는 물질을 생산하기 위하여 대전광역시 월평공원으로부터 분리 동정한 야생효모들 중 비병원성 야생효모들을 선별하여 배양 상등액과 무세포 추출물을 각각 제조한 후 β-glucuronidase 저해활성을 측정하였다. 133균주의 비병원성 야생효모들 중 C. oleophila WP5-19-1의 무세포추출물이 49.0%의 가장 높은 저해활성을 보여 우수 균주로 최종 선발하였다. β-Glucuronidase 저해물질 최적 생산 조건은 C. oleophila WP5-19-1를 초기 pH를 6.0으로 조정한 dextrose 5% 함유한 potato dextrose 배지에 접종하여 30℃에서 24시간 배양하였을 때였고 이때 IC50값; 8.4 mg으로 가장 높은 저해활성을 보였다. C. oleophila WP5-19-1의 β-glucuronidase 저해물질을 한외여과와 Sephadex G-50으로 gel 여과를 실시하여 부분 정제하였고 이들은 30-60℃, pH 6.0-9.0에서 80% 이상의 잔존 저해활성을 보여 안정하였다. 또한 철, 구리, 아연과 같은 중금속 이온들은 저해활성을 활성화시켰다.

Keywords

References

  1. Kim DH, Choi HJ, Bae EA, Han MJ, Park SY. Effect of artificially cultured Phellinus linteus on harmful intestinal bacterial enzymes and rat intestinal α-glucosidases. J Fd Hyg Safety 1998;13:20-3. 
  2. Lee KA, Kim MS, Cho HB. Effect of extract of fermented dropwort on intestinal bacteria and enzymes in vitro. Kor J Microbiol 2008;44:358-61. 
  3. Lee HC, Andrew MJ, Low CS, Lee YK. Effect of tea phenolics and their aromatic fecal bacterial metabolites on intestinal microbiota. Research in Microbiol 2006;157:876-84.  https://doi.org/10.1016/j.resmic.2006.07.004
  4. Rhee YK, Kim DH, Han MJ. Inhibitory effect of Zizyphi fructus on β-glucuronidase and tryptophanase of human intestinal bacteria. Kor J Food Sci Technol 1998;30:199-205. 
  5. Myung DS, Joo YE. Gut microbial influence and probiotics on colorectal cancer. Korean J Gastronterol 2012;60:275-84.  https://doi.org/10.4166/kjg.2012.60.5.275
  6. Kim EJ, Shin IU, Kwun SY, Park EH, Yi JH, Kim MD. Exploration of β-glucuronidase activity of lactic acid bacteria isolated from Kimchi. Microbiol Biotechnol Lett 2019;47:434-40.  https://doi.org/10.4014/mbl.1901.01002
  7. Kim KY, Choi KS, Hideyuki K, Kim SM. β-Glucuronidase inhibitory activity of bromophenols purified from Grateloupia elliptica. Food Sci Biotechnol 2008;17:1110-4. 
  8. Han MJ, Bae EA, Rhee YK, Kim DH. Effect of mushrooms on the growth of intestinal lactic acid bacteria. Korean J of Food Sci Technol 1996;28:947-52. 
  9. Weng ZM, Wang P, Ge GB, D ZR, Wu DC, Zou LW, Dou TY, Hou J, Zhang TY, Yang L. Structure-activity relationships of flavonoids as natural inhibitors against E.coli β-glucuronidase. Food Chem Toxicol 2017;109:975-83.  https://doi.org/10.1016/j.fct.2017.03.042
  10. Bai Y, Chen L, Cao YF, Hou XD, Jia SN, Zhou Q, He YQ, Hou J. Beta-glucuronidase inhibition by constituents of mulberry bark. Planta Med 2021;87:631-41.  https://doi.org/10.1055/a-1402-6431
  11. Kim JY, Han SM, Lee JS. Isolation and tyrosinase inhibitory activity of wild yeasts obtained from soil in the fields of medicinal plants, ginseng and Korean angelica. Kor J Mycol 2018;46:315-323. 
  12. Kim JH, Lee DH, Jeong SC, Chung KS, Lee JS. Characterization of antihypertensive angiotensin I-converting enzyme inhibitor from Saccharomyces cerevisiae. J Microbiol Biotechnol 2004;14:1318-23. 
  13. Lee DH, Lee DH, Lee JS. Characterization of a new antidementia β-secretase inhibitory peptide from Saccharomyces cerevisiae. Enzyme Microb Technol 2007;42:83-8.  https://doi.org/10.1016/j.enzmictec.2007.08.003
  14. Lee DH, Lee JS, Yi SH, Lee JS. Production of the acetylcholinesterase inhibitor from Yarrowia lipolytica S-3. Mycobiol 2008;36:102-5.  https://doi.org/10.4489/MYCO.2008.36.2.102
  15. Jeong SC, Lee DH, Lee JS. Production and characterization of an anti-angiogenic agent from Saccharomyces cerevisiae K-7. J Micriobiol Biotechnol 2006;16:1904-11. 
  16. Jang IT, Kim YH, Kang MG, Yi SH, Lim SI, Lee JS. Production of tyrosinase inhibitor from Saccharomyces cerevisiae. Kor J Mycol 2012;40:60-4.  https://doi.org/10.4489/KJM.2012.40.1.060
  17. Jang IT, Kim YH, Yi SH, Lim SI, Lee JS. Screening of a new fibrinolytic substances-producing yeast. Kor J Mycol 2011;39:227-8.  https://doi.org/10.4489/KJM.2010.39.3.227
  18. Kim HK, Kim JY, Han SM, Kim CM, Lee JS. Microbiological characteristics and physiological functionalities of unrecorded wild yeast strains in the soils of Hajodae and Gyungpodae beaches in Korea. Kor J Mycol 2019;47:249-58. 
  19. Han SM, Kim JY, Kim C, Lee JS. Characteristics of unrecorded wild yeasts obtained from soil of spices plant fields and its physiological functionality. Kor J Mycol 2019;47:75-81. 
  20. Lee JS, Yi SH, Kwon SJ, Ahn C, Yoo JY. Microbiological characteristics and physiological functionality of unrecorded yeasts from mountains soils in Daejeon metropolitan city and Chungcheongnam-do, Korea. Kor J Mycol 2016;44:138-44. 
  21. Jeong SC, Hyun KW, Kim JH, Lee JS. Isolation of a halotolerant yeast and the production of extracellular protease. Kor J Biotechnol Bioeng 2001;16:158-62. 
  22. Jang JE, Park SJ, Lee JS. Isolation of wild yeasts and characterization of physiological functionalities of unrecorded wild yeasts obtained from flowers and soils of the Wolpyung park, Daejeon city and Gykpo beach of Jeollabuk-do in Korea. Kor J Mycol 2021;49:87-100. 
  23. Kim JH, Lee DH, Jeong SC, Chung KS, Lee JS. Characterization of antihypertensive angiotensin I-converting enzyme inhibitor from Saccharomyces cerevisae. J Microbiol Biotechnol 2004;14:1318-23. 
  24. Oh EH, Park JM, Kim SH, Song IG, Han NS, Yoon HS. Biological activities of Phellinus linteus mycelium culture with cassia semen extract on β-glucuronidase inhibitory activity. Kor J Food & Nutr 2012;25:620-8.  https://doi.org/10.9799/ksfan.2012.25.3.620
  25. Yoo J, Hwang JS, Goo TW, Yun EY. Comparative analysis of nutritional and harmful components in Korean and Chinese mealworms (Tenebrio molitor). J Kor Soc Food Sci Nutr 2013;42;249-54. https://doi.org/10.3746/jkfn.2013.42.2.249