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Synthesis of raffinose by fungal α-galacotosidase from Absidia corymbifera

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Abstract

In order to investigate the optimal conditions for raffinose synthesis, α-galactosidase was purified from Absidia corymbifera IFO8084 with a recovery yield of approximately 8.1% (8.36 mg). The molecular weight of the wild-type α-galactosidase was about 83 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The native molecular mass of the enzyme was approximately 330 kDa by gel filtration chromatography, indicating that α-galactosidase from A. corymbifera IFO8084 is a homotetrameric enzyme. The purified enzyme displayed optimal enzyme activity at pH 4.5 and 60°C. When the purified α-galactosidase was incubated in a substrate solution of sucrose and D-galactose for 48 hr at 37°C, raffinose was synthesized and was confirmed by thin layer chromatography (TLC), high performance liquid chromatography (HPLC), and 13C-nuclear magnetic resonance (13C-NMR) spectrometer analysis. Maximum rates of conversion were observed with 1.67 M galactose, 2.04 M sucrose, and 100 U α-galactosidase at pH 6.0 and 70°C. Under the optimized conditions, the overall conversion ratio was 10%(w/v), representing 2.5 times the synthesis yield that would be possible without the optimized conditions.

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References

  1. Ballongue J, Bifidobacteria and probiotic action. pp. 357–442. In: Lactic Acid Bacteria, Microbiology, and Functional Aspects. Salminen S, von Wright A (eds). Marcel Dekker, Inc, New York, NY, USA (1998)

    Google Scholar 

  2. Mituoka T. Bifidobacteria and their role in human health. J. Ind. Microbiol. 6: 263–268 (1990)

    Article  Google Scholar 

  3. Gibson GR, Roberfroid MB. Dietary modulation of human colonic microbiota: Introducing the concept of prebiotic action. J. Nutr. 125: 1401–1412 (1995)

    CAS  Google Scholar 

  4. Mitsuoka T, Hidaka H, Eida T. Effect of fructooligosaccharides on intestinal microflora. Nahrung 31: 5–6 (1987)

    Article  Google Scholar 

  5. Lehle L, Tanner W. The function of myo-inositol in the biosynthesis of raffinose. Eur. J. Biochem. 38: 103–110 (1973)

    Article  CAS  Google Scholar 

  6. Schmidt RR. New methods for the synthesis of glycosides and oligosaccharides. Are there alternatives to the Koenigs-Knorr methods? Angew. Chem. Int. Edit. 25: 212–235 (1986)

    Article  Google Scholar 

  7. Wong CH, Haynie SL, Whitesides GM. Enzyme-catalyzed synthesis of N-acetu;actpsa, ome wotj in situ regeneration of uridine 5′-diphosphate glucose and uridine 5′-diphosphate galactose. J. Org. Chem. 47: 5416–5418 (1982)

    Article  CAS  Google Scholar 

  8. Lowry OH, Farr NJ, Randall RL. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265–275 (1951)

    CAS  Google Scholar 

  9. Laemmli UK. Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970)

    Article  CAS  Google Scholar 

Download references

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  1. Department of Food Science and Human Nutrition, and Research Institute of Human Ecology, Chonbuk National University, Jeonju, Jeonbuk, 561-756, Korea

    Sang-Ho Baik

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  1. Sang-Ho Baik
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Correspondence to Sang-Ho Baik.

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Baik, SH. Synthesis of raffinose by fungal α-galacotosidase from Absidia corymbifera . Food Sci Biotechnol 19, 83–87 (2010). https://doi.org/10.1007/s10068-010-0012-3

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  • DOI: https://doi.org/10.1007/s10068-010-0012-3

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