Optimization of Ascorbic Acid-2-Phosphate Production from Ascorbic Acid Using Resting Cell of Brevundimonas diminuta

  • Shin, Woo-Jung (Department of Biotechnology and Genetic Engineering, College of Life and Environmental Sciences, Korea University) ;
  • Kim, Byung-Yong (Korean Agricultural Culture Collection(KACC), Microbial Genetics Division, National Institute of Agricultural Biotechnology) ;
  • Bang, Won-Gi (Department of Biotechnology and Genetic Engineering, College of Life and Environmental Sciences, Korea University)
  • Published : 2007.05.31

Abstract

With the aim to produce ascorbic acid-2-phosphate(AsA-2-P) from L-ascorbic acid(AsA, Vitamin C), nine bacteria conferring the ability to transform AsA to AsA-2-P were isolated from soil samples alongside known strains from culture collections. Most isolates were classified to the genus Brevundimonas by 16S phylogenetic analysis. Among them, Brevundimonas diminuta KACC 10306 was selected as the experimental strain because of its the highest productivity of AsA-2-P. The optimum set of conditions for the AsA-2-P production from AsA using resting cells as the source of the enzyme was also investigated. The optimum cultivation time was 16 h and the cell concentration was 120g/l(wet weight). The optimum concentrations of AsA and pyrophosphate were 550mM and 450mM, respectively. The most effective buffer was 50mM sodium formate. The optimum pH was 4.5 and temperature was $40^{\circ}C$. Under the above conditions, 27.5g/l of AsA-2-P was produced from AsA after 36 h of incubation, which corresponded to a 19.7% conversion efficiency based on the initial concentration of AsA.

Keywords

References

  1. Colven, R. M. and S. R. Pinnell. 1996. Topical vitamin C in aging. Clin. Dermatol. 14: 227-234 https://doi.org/10.1016/0738-081X(95)00158-C
  2. Felsenstein, J. 1985. Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39: 783-791 https://doi.org/10.2307/2408678
  3. Geesin, J. C., J. S. Gordon, and R. A. Berg. 1993. Regulation of collagen synthesis in human dermal fibroblasts by the sodium and magnesium salts of ascorbyl-2-phosphate. Skin Pharmacol. 6: 65-71 https://doi.org/10.1159/000211089
  4. Hata, R. and H. Senoo. 1989. L-Ascorbic acid 2-phosphate stimulates collagen accumulation, cell proliferation, and formation of a three-dimensional tissuelike substance by skin fibroblasts. J. Cell. Physiol. 138: 8-16 https://doi.org/10.1002/jcp.1041380103
  5. Huang, A., J. A. Vita, R. C. Venema, and J. F. Keaney Jr. 2000. Ascorbic acid enhances endothelial nitric-oxide synthase activity by increasing intracellular tetrahydrobiopterin. J. Biol. Chem. 275: 17399-17406 https://doi.org/10.1074/jbc.M002248200
  6. Kim, B. Y., S. Y. Lee, H. Y. Weon, S. W. Kwon, S. J. Go, Y. K. Park, P. Schumann, and D. Fritze. 2006. Ureibacillus suwonensis sp. nov., isolated from cotton waste composts. Int. J. Syst. Evol. Microbiol. 56: 663-666 https://doi.org/10.1099/ijs.0.63703-0
  7. Kim, T. W., S. W. Jung, Y. H. Go, and Y. H. Lee. 2006. Novel suspension-phase enzyme reaction system using insoluble extrusion starch as glycosyl donor for intermolecular transglycosylation of L-ascorbic acid. J. Microbiol. Biotechnol. 16: 1678-1683
  8. Kimura, M. 1983. The Neutral Theory of Molecular Evolution. Cambridge University Press, Cambridge
  9. Kobayashi, S., M. Takehana, S. Itoh, and E. Ogata. 1996. Protective effect of magnesium-L-ascorbyl-2-phosphate against skin damage induced by UVB irradiation. Photochem. Photobiol. 64: 224-228 https://doi.org/10.1111/j.1751-1097.1996.tb02447.x
  10. Kumar, S., K. Tamura, and M. Nei. 2004. MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief. Bioinform. 5: 150-163 https://doi.org/10.1093/bib/5.2.150
  11. Kwon, K. S., S. H. Lee, and W. G. Bang. 2000. Production of ascorbic acid-2-phosphate from ascorbic acid by Pseudomonas sp. Korean J. Appl. Microbiol. Biotechnol. 28: 33-38
  12. Lee, C. Y., K. H. Kim, S. Y. Hur, J. H. Heo, and M. H. Choi. 2006. Enzymatic synthesis of ascorbic acid fructoside by transfructosylation using levan fructotransferase. J. Microbiol. Biotechnol. 16: 64-67
  13. Mima, H., H. Nomura, Y. Imai, and H. Takashima. 1970. Chemistry and application of ascorbic acid phosphate. Vitamins Japan 41: 387-398
  14. Nomura, H. and S. Morimoto. 1971. Chem. Pharm. Bull. 19: 335 https://doi.org/10.1248/cpb.19.335
  15. Ono, M., Y. Aratani, I. Kitagawa, and Y. Kitagawa. 1990. Ascorbic acid phosphate stimulates type IV collagen synthesis and accelerates adipose conversion of 3T3-L1 cell. Exp. Cell. Res. 187: 309-314 https://doi.org/10.1016/0014-4827(90)90096-S
  16. Saika, S., R. Kanagawa, K. Uenoyama, K. Hiroi, and J. Hiraoka. 1991. L-Ascorbic acid 2-phosphate, a phosphate derivative of L-ascorbic acid, enhances the growth of cultured rabbit keratocytes. Graefes Arch. Clin. Exp. Ophthalmol. 229: 79-83 https://doi.org/10.1007/BF00172267
  17. Saitou, N. and M. Nei. 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molec. Biol. Evol. 4: 406-425
  18. Sauberlich, H. E. 1994. Pharmacology of vitamin C. Annu. Rev. Nuctr. 14: 371-391 https://doi.org/10.1146/annurev.nu.14.070194.002103
  19. Sekine M., T. Futatsugi, and T. Hata. 1982. Silyl phosphites. 21. A new method for the synthesis of L-ascorbic acid 2-Ophosphate by utilizing phosphoryl rearrangement. J. Org. Chem. 47: 3453-3456 https://doi.org/10.1021/jo00139a014
  20. Takamura, K. and M. Ito. 1997. Effects of metal ions and flavonoids on the oxidation of ascorbic acid. Chem. Pharm. 25: 3218-3225
  21. Takebayashi, J., A. Tai, E. Gohda, and I. Yamamoto. 2006. Characterization of the radical-scavenging reaction of 2-Osubstituted ascorbic acid derivatives, AA-2G, AA-2P, and AA-2S: A kinetic and stoichiometric study. Biol. Pharm. Bull. 29: 766-771 https://doi.org/10.1248/bpb.29.766
  22. Thompson, J. D., D. G.. Higgins, and T. J. Gibson. 1994. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22: 4673-4680 https://doi.org/10.1093/nar/22.22.4673