DOI QR코드

DOI QR Code

Production of Deglucose-ApioseXylosylated Platycosides from Glycosylated Platycosides by Crude Enzyme from Aspergillus tubingensis

  • Shin, Kyung-Chul (Department of Integrative Bioscience and Biotechnology, Konkuk University) ;
  • Kil, Tae-Geun (Department of Bioscience and Biotechnology, Konkuk University) ;
  • Kang, Su-Hwan (Department of Bioscience and Biotechnology, Konkuk University) ;
  • Oh, Deok-Kun (Department of Integrative Bioscience and Biotechnology, Konkuk University)
  • Received : 2021.12.10
  • Accepted : 2022.02.15
  • Published : 2022.04.28

Abstract

Platycosides, Platycodi radix (Platycodon grandiflorus root) saponins, are used as food supplements and exert diverse pharmacological activities. Deglycosylation of saponins enhances their biological efficacy, and deglycosylated platycosides are produced mainly through enzymatic hydrolysis. However, the types of available deglycosylated platycosides remain limited because of a lack of hydrolyzing enzymes that can act on specific glycosides in glycosylated platycosides. In this study, a crude enzyme from Aspergillus tubingensis converted platycoside E (PE) and polygalacin D3 (PGD3) into deglucose-apiose-xylosylated (deGAX)-platycodin D (PD) and deGAX-polygalacin D (PGD), respectively. The products were identified through LC/MS analysis by specifically hydrolyzing all glucose residues at C-3, and apiose and xylose residues at C-28 of platycoside. The hydrolytic activity of the crude enzyme obtained after the cultivation of the fungus using citrus pectin and corn steep solid as carbon and nitrogen sources, respectively, in culture medium was increased compared with those using other carbon and nitrogen sources. The crude enzyme from A. tubingensis was the most effective in producing deGAX platycoside at pH 5.0 and 60℃. The crude enzyme produced 0.32 mg/ml deGAX-PD and 0.34 mg/ml deGAX-PGD from 1 mg/ml PE and 1 mg/ml PGD3 (at pH 5.0 and 60℃) for 12 and 10 h, with productivities of 32.0 and 42.5 mg/l/h and molar yields of 62.1 and 59.6%, respectively. To the best of our knowledge, this is the first study to produce deGAX platycosides from glycosylated platycosides.

Keywords

Acknowledgement

This study was supported by the KU Research Professor Program of Konkuk University.

References

  1. Nyakudya E, Jeong JH, Lee NK, Jeong YS. 2014. Platycosides from the roots of Platycodon grandiflorum and their health benefits. Prev. Nutr. Food Sci. 19: 59-68. https://doi.org/10.3746/PNF.2014.19.2.059
  2. Kim JY, Hwang YP, Kim DH, Han EH, Chung YC, Roh SH, et al. 2006. Inhibitory effect of the saponins derived from roots of Platycodon grandiflorum on carrageenan-induced inflammation. Biosci. Biotechnol. Biochem. 70: 858-864. https://doi.org/10.1271/bbb.70.858
  3. Kim M, Hwang IG, Kim SB, Choi AJ. 2020. Chemical characterization of balloon flower (Platycodon grandiflorum) sprout extracts and their regulation of inflammatory activity in lipopolysaccharide-stimulated RAW 264.7 murine macrophage cells. Food Sci. Nutr. 8: 246-256. https://doi.org/10.1002/fsn3.1297
  4. Ryu CS, Kim CH, Lee SY, Lee KS, Choung KJ, Song GY, et al. 2012. Evaluation of the total oxidant scavenging capacity of saponins isolated from Platycodon grandiflorum. Food Chem. 132: 333-337. https://doi.org/10.1016/j.foodchem.2011.10.086
  5. Luo H, Lin S, Ren F, Wu L, Chen L, Sun Y. 2007. Antioxidant and antimicrobial capacity of Chinese medicinal herb extracts in raw sheep meat. J. Food Prot. 70: 1440-1445. https://doi.org/10.4315/0362-028X-70.6.1440
  6. Xie Y, Ye YP, Sun HX, Li D. 2008. Contribution of the glycidic moieties to the haemolytic and adjuvant activity of platycodigenin-type saponins from the root of Platycodon grandiflorum. Vaccine 26: 3452-3460. https://doi.org/10.1016/j.vaccine.2008.04.023
  7. Noh EM, Kim JM, Lee HY, Song HK, Joung SO, Yang HJ, et al. 2019. Immuno-enhancement effects of Platycodon grandiflorum extracts in splenocytes and a cyclophosphamide-induced immunosuppressed rat model. BMC Complement. Altern. Med. 19: 322. https://doi.org/10.1186/s12906-019-2724-0
  8. Zhao X, Wang Y, Yan P, Cheng G, Wang C, Geng N, et al. 2017. Effects of polysaccharides from Platycodon grandiflorum on immunity-enhancing activity in vitro. Molecules 22: 1918. https://doi.org/10.3390/molecules22111918
  9. Yim NH, Hwang YH, Liang C, Ma JY. 2016. A platycoside-rich fraction from the root of Platycodon grandiflorum enhances cell death in A549 human lung carcinoma cells via mainly AMPK/mTOR/AKT signal-mediated autophagy induction. J. Ethnopharmacol. 194: 1060-1068. https://doi.org/10.1016/j.jep.2016.10.078
  10. Kim YS, Kim JS, Choi SU, Kim JS, Lee HS, Roh SH, et al. 2005. Isolation of a new saponin and cytotoxic effect of saponins from the root of Platycodon grandiflorum on human tumor cell lines. Planta Med. 71: 566-568. https://doi.org/10.1055/s-2005-864161
  11. Khan M, Maryam A, Zhang H, Mehmood T, Ma T. 2016. Killing cancer with platycodin D through multiple mechanisms. J. Cell Mol. Med. 20: 389-402. https://doi.org/10.1111/jcmm.12749
  12. Han LK, Zheng YN, Xu BJ, Okuda H, Kimura Y. 2002. Saponins from platycodi radix ameliorate high fat diet-induced obesity in mice. J. Nutr. 132: 2241-2245. https://doi.org/10.1093/jn/132.8.2241
  13. Zhao HL, Harding SV, Marinangeli CP, Kim YS, Jones PJ. 2008. Hypocholesterolemic and anti-obesity effects of saponins from Platycodon grandiflorum in hamsters fed atherogenic diets. J. Food Sci. 73: H195-200. https://doi.org/10.1111/j.1750-3841.2008.00915.x
  14. Hwang KA, Hwang YJ, Im PR, Hwang HJ, Song J, Kim YJ. 2019. Platycodon grandiflorum extract reduces high-fat diet-induced obesity through regulation of adipogenesis and lipogenesis pathways in mice. J. Med. Food 22: 993-999. https://doi.org/10.1089/jmf.2018.4370
  15. Ha YW, Na YC, Seo JJ, Kim SN, Linhardt RJ, Kim YS. 2006. Qualitative and quantitative determination of ten major saponins in Platycodi Radix by high performance liquid chromatography with evaporative light scattering detection and mass spectrometry. J. Chromatogr. A. 1135: 27-35. https://doi.org/10.1016/j.chroma.2006.09.015
  16. Yoo DS, Choi YH, Cha MR, Lee BH, Kim SJ, Yon GH, et al. 2011. HPLC-ELSD analysis of 18 platycosides from balloon flower roots (Platycodi Radix) sourced from various regions in Korea and geographical clustering of the cultivation areas. Food Chem. 129: 645-651. https://doi.org/10.1016/j.foodchem.2011.04.106
  17. Park CS, Yoo MH, Noh KH, Oh DK. 2010. Biotransformation of ginsenosides by hydrolyzing the sugar moieties of ginsenosides using microbial glycosidases. Appl. Microbiol. Biotechnol. 87: 9-19. https://doi.org/10.1007/s00253-010-2567-6
  18. Shin KC, Oh DK. 2016. Classification of glycosidases that hydrolyze the specific positions and types of sugar moieties in ginsenosides. Crit. Rev. Biotechnol. 36: 1036-1049. https://doi.org/10.3109/07388551.2015.1083942
  19. Kim WY, Kim JM, Han SB, Lee SK, Kim ND, Park MK, et al. 2000. Steaming of ginseng at high temperature enhances biological activity. J. Nat. Prod. 63: 1702-1704. https://doi.org/10.1021/np990152b
  20. Zhou W, Yan Q, Li JY, Zhang XC, Zhou P. 2008. Biotransformation of Panax notoginseng saponins into ginsenoside compound K production by Paecilomyces bainier sp. 229. J. Appl. Microbiol. 104: 699-706. https://doi.org/10.1111/j.1365-2672.2007.03586.x
  21. Jiang Y, Li W, Fan D. 2021. Biotransformation of ginsenoside Rb1 to ginsenoside CK by strain XD101: a safe bioconversion strategy. Appl. Biochem. Biotechnol. 193: 2110-2127. https://doi.org/10.1007/s12010-021-03485-0
  22. Shin KC, Kim DW, Oh YJ, Seo MJ, Na CS, Kim YS. 2021. Improved production of deglucosylated platycodin D from saponins from balloon flower leaf by a food-grade enzyme using high hydrostatic pressure. Heliyon 7: e08104. https://doi.org/10.1016/j.heliyon.2021.e08104
  23. Ahn HJ, You HJ, Park MS, Johnston TV, Ku S, Ji GE. 2018. Biocatalysis of platycoside E and platycodin D3 using fungal extracellular β-glucosidase responsible for rapid platycodin D production. Int. J. Mol. Sci. 19: 2671. https://doi.org/10.3390/ijms19092671
  24. Shin KC, Seo MJ, Kim DW, Yeom SJ, Kim YS. 2019. Characterization of β-glycosidase from Caldicellulosiruptor owensensis and its application in the production of platycodin D from balloon flower leaf. Catalysts 9: 1025. https://doi.org/10.3390/catal9121025
  25. Kil TG, Kang SH, Kim TH, Shin KC, Oh DK. 2019. Enzymatic biotransformation of balloon flower root saponins into bioactive platycodin D by deglucosylation with Caldicellulosiruptor bescii β-glucosidase. Int. J. Mol. Sci. 20: 3854. https://doi.org/10.3390/ijms20163854
  26. Kang SH, Kim TH, Shin KC, Ko YJ, Oh DK. 2019. Biotransformation of food-derived saponins, platycosides, into deglucosylated saponins Including deglucosylated platycodin D and their anti-inflammatory activities. J. Agric. Food Chem. 67: 1470-1477. https://doi.org/10.1021/acs.jafc.8b06399
  27. Jeong EK, Ha IJ, Kim YS, Na YC. 2014. Glycosylated platycosides: identification by enzymatic hydrolysis and structural determination by LC-MS/MS. J. Sep. Sci. 37: 61-68. https://doi.org/10.1002/jssc.201300918
  28. Li W, Zhao LC, Wang Z, Zheng YN, Liang J, Wang H. 2012. Response surface methodology to optimize enzymatic preparation of deapio-platycodin D and platycodin D from radix platycodi. Int. J. Mol. Sci. 13: 4089-4100. https://doi.org/10.3390/ijms13044089
  29. Ha IJ, Ha YW, Kang M, Lee J, Park D, Kim YS. 2010. Enzymatic transformation of platycosides and one-step separation of platycodin D by high-speed countercurrent chromatography. J. Sep. Sci. 33: 1916-1922. https://doi.org/10.1002/jssc.200900842
  30. Wie HJ, Zhao HL, Chang JH, Kim YS, Hwang IK, Ji GE. 2007. Enzymatic modification of saponins from Platycodon grandiflorum with Aspergillus niger. J. Agric. Food Chem. 55: 8908-8913. https://doi.org/10.1021/jf0716937
  31. Shin KC, Kil TG, Lee TE, Oh DK. 2021. Production of bioactive beapiosylated platycosides from glycosylated platycosides in balloon flower root using the crude enzyme from the food-available fungus Rhizopus oryzae. J. Agric. Food Chem. 69: 4766-4777. https://doi.org/10.1021/acs.jafc.0c06756
  32. Shin KC, Kim DW, Woo HS, Oh DK, Kim YS. 2020. Conversion of glycosylated platycoside E to deapiose-xylosylated platycodin D by cytolase PCL5. Int. J. Mol. Sci. 21: 1207. https://doi.org/10.3390/ijms21041207
  33. Kim SA, Jeong EB, Oh DK. 2021. Complete bioconversion of protopanaxadiol-type ginsenosides to compound k by extracellular enzymes from the isolated strain Aspergillus tubingensis. J. Agric. Food Chem. 69: 315-324. https://doi.org/10.1021/acs.jafc.0c07424
  34. Jeong EB, Kim SA, Shin KC, Oh DK. 2020. Biotransformation of protopanaxadiol-type ginsenosides in Korean ginseng extract into food-available compound K by an extracellular enzyme from Aspergillus niger. J. Microbiol. Biotechnol. 30: 1560-1567. https://doi.org/10.4014/jmb.2007.07003
  35. Lara-Espinoza C, Carvajal-Millan E, Balandran-Quintana R, Lopez-Franco Y, Rascon-Chu A. 2018. Pectin and pectin-based composite materials: beyond food texture. Molecules 23: 942. https://doi.org/10.3390/molecules23040942