Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Partial Purification and Characterization of Limonoate Dehydrogenase from Rhodococcus fascians for the Degradation of Limonin

  • Published : 2002.08.01
Download PDF
⟨ Previous Next ⟩

Abstract

An extracellular limonoate dehydrogenase was purified 10-fold from a cell-free extract of Rhodococcus fascians by ammonium sulfate precipitation, dialysis, and ultrafiltration. This purified dehydrogenase catalyzed the conversion of limonoate to 17-dehydrolimonoate. The enzyme showed optimum activity at pH 8.0 and $40^{\circ}C$, with $K_m$ value of 0.9$\muM$, and requires Zn ions and sulfhydryl groups for catalytic action. The enzyme activity was inhibited by $Hg^{2+}\;and\;NaN_3$ ions. The degradation of limonin (66%) in Kinnow mandarin juice was successfully demonstrated with partially purified limonoate dehydrogenase. With scale-up preparation of limonoate dehydrogenase, a successful debittering operation of fruit juices appears feasible.

Keywords

References

  1. Official Methods of Analysis(14th ed.) AOAC
  2. Indian Food Packers v.12 Studies of bitterness in orange cultivars Bhalerao, S. D.;G. V. Mulmuley
  3. Anal. Chem. v.31 Determination of flavonones in citrus fruits Davis, W. B. https://doi.org/10.1021/ac60147a030
  4. J. Sci. Food Agric. v.40 A review of limonin in grapefruit juice, its relationship to flavor and efforts to reduce it Fellers, P. J.
  5. J. Agric. Food Chem. v.20 Limonoate dehydrogenase from Arthrobacter globiformis Hasegawa, S.;R. D. Bennett;V. P. Maier;A. D. King https://doi.org/10.1021/jf60183a001
  6. J. Food Sci. v.38 Use of limonoate dehydrogenase of Arthrobacter globiformis for the prevention or removal of limonin bitterness in citrus products Hasegawa, S.;L. C. Brewster;V. P. Maier https://doi.org/10.1111/j.1365-2621.1973.tb07225.x
  7. J. Agric. Food Chem. v.31 A species of bacterium producing constitutive enzymes for limonoid metabolism Hasegawa, S.;A. D. King https://doi.org/10.1021/jf00118a032
  8. Bitterness in Foods and Beverages Biochemistry of limonid citrus juice bitter principles and biochemical debittering processes Hasegawa, S.;V. P. Maier;R. L. Rouseff(ed.)
  9. Appl. Microbiol. Biotechnol. v.41 Continuous limonin degradation by immobilized Rhodococcus fascians cells k-carragenaan Iborra, J. L.;A. Manjon;M. Canovas;P. Lazano;C. Martinez
  10. Punjab Horticulture J. v.13 Kinnow holds out promise for Punjab Jawanda, J. S.;K. Singh
  11. J. Biol. Chem. v.193 Protein measurement with Folin phenol reagent Lowry, O. H.;A. L. Rosebrough;A. L. Farr;R. J. Randall
  12. Phytochemistry v.8 Limonin-D-ring lactone hydrolase. A new enzyme from Citrus seeds Maier, V. P.;S. Hasegawa;E. Hera https://doi.org/10.1016/S0031-9422(00)85439-4
  13. Anal. Chem. v.31 Use of dinitrosalicylic acid reagent for determination of reducing sugars Miller, G. L. https://doi.org/10.1021/ac60147a030
  14. Enz. Microb. Technol. v.16 Optimization of parameters for hydrolysis of limonin for debittering of Kinnow mandarin juice by Rhodococcus fascians Marwaha, S. S.;M. Puri;M. Bhullar;R. M. Kothari https://doi.org/10.1016/0141-0229(94)90097-3
  15. PhD thesis, Punjabi University Immobilized enzyme technology for the clarification and debittering of citrus fruit juices with special reference to Kinnows Puri, M.
  16. CRC Rev. Biotechnol. v.16 Biochemical basis of bitterness in citrus fruit juices and biotech approaches for debittering Puri, M.;S. S. marwaha;R. M. Kothari;J. F. Kennedy https://doi.org/10.3109/07388559609147419
  17. Enz. Microbiol. Technol. v.18 Studies on the applicability of alginate entrapped naringinase for the debittering of Kinnow juice Puri, M.;S. S. Marwaha;R. M. Kothari https://doi.org/10.1016/0141-0229(95)00100-X
  18. Biotechnol. Adv. v.18 Production, purification and characterization of the debittering enzyme naringinase Puri, M.;U. C. Banerjee https://doi.org/10.1016/S0734-9750(00)00034-3
  19. Food Biotechnol. v.15 Debittering of kinnow mandarin juice by covalently bound naringinase on hen egg white Puri, M.;M. Seth;S. S. Marwaha;R. M. Kothari https://doi.org/10.1081/FBT-100103891
  20. Anal. Chem. v.52 Determination of limonin in citrus juices by high pressure liquid chromatography Rouseff, R. L.;J. F. Fischer https://doi.org/10.1021/ac50058a017
  21. Indian Farming Kinnow orange in juice and beverage making Sethi, V;J. C. Anand;S. K. Saxena
  22. J. Agric. Food Chem. v.6 Enzymatic hydrolysis of naringin in grapefruit Ting, S. V. https://doi.org/10.1021/jf60089a010
  23. J. Agric. Food Chem. v.29 Debittering of orange juice by bacteria which degrade limonin Vaks, B.;A. Lifshitz https://doi.org/10.1021/jf00108a039