Naringinases: occurrence, characteristics, and applications

  • Maria H. RibeiroEmail author


Naringinase, an enzyme complex, is commercially attractive due to its potential usefulness in pharmaceutical and food industries. It is of particular interest in the biotransformation of steroids, antibiotics, and mainly of glycosides hydrolysis. Moreover, it can be used in citrus juices debittering and wine industries. Naringinase expresses activity on α-l-rhamnosidase and β-d-glucosidase. Many natural glycosides, including naringin, rutin, quercitrin, hesperidin, diosgene, and ter-phenyl glycosides, containing terminal α-rhamnose and β-glucose can act as substrates of naringinase. The sources, production, activity, biochemical properties, and substrate specificity of naringinase are reviewed, along with a description of the enzymatic deglycosylation systems and applications, concluding with the identification of areas which need further extensive studies.


Naringinase α-l-Rhamnosidase β-d-Glucosidase Glycosides Deglycosylation Immobilization 


  1. Amaro MI, Rocha J, Vila-Real H, Eduardo-Figueira M, Mota-Filipe H, Sepodes B, Mota-Filipe H, Ribeiro MHL (2009) Anti-inflamatory activity of naringin and the biosynthesised naringenin by naringinase immobilised in microstructure materials in a model DSS-induced colitis in mice. Food Res Int 42:1010–1017CrossRefGoogle Scholar
  2. Birgisson H, Wheat JO, Hreggvidsson GO, Kristjansson JK, Mattiasson B (2007) Immobilization of a recombinant Escherichia coli producing a thermostable rhamnosidase: creation of a bioreactor for hydrolyses of naringin. Enz Microb Technol 40:1181–1187CrossRefGoogle Scholar
  3. Bram B, Solomons GL (1965) Production of the enzyme naringinase by Aspergillus niger. Appl Microbiol 13:842–845Google Scholar
  4. Busto MD, Meza V, Ortega N, Perez-Mateos M (2007) Immobilization of naringinase from Aspergillus niger CECT 2088 in poly(vinyl alcohol) cryogels for the debittering of juices. Food Chem 104:1177–1182CrossRefGoogle Scholar
  5. Caldini C, Bonomi F, Pifferi PG, Lanzarini G, Galente YM (1994) Kinetic and immobilization studies on fungal glycosidase for aroma enhancement in wine. Enz Microb Technol 16:286–291CrossRefGoogle Scholar
  6. Chang H-Y, Lee Y-B, Bae H-A, Huh J-Y, Nam S-H, Sohn H-S, Lee HJ, Lee S-B (2011) Purification and characterisation of Aspergillus sojae naringinase: the production of prunin exhibiting markedly enhanced solubility with in vitro inhibition of HMG-CoA reductase. Food Chem 124:234–241CrossRefGoogle Scholar
  7. Chen Y, Shen S, Lin H (2003) Rutinoside at C7 attenuates the apoptosis-inducing activity of flavonoids. Biochem Pharm 66:1139–1150CrossRefGoogle Scholar
  8. Chien PJ, Fuu S, Shyu YT (2001) Monitoring enzymatic debittering in grapefruit juice by high performance liquid chromatography. J Food Drug Anal 9:115–120Google Scholar
  9. Daniels L, Linhardt RJ, Bryan BA, Mayerl F, Pickenhagen M (1990) Methods for producing rhamnose. US Patent 4(933):281Google Scholar
  10. Davis DW (1947) Determination of flavonones in citrus juice. Anal Chem 19:46–48CrossRefGoogle Scholar
  11. Dunlap WJ, Hagen RE, Wender SH (1962) Preparation and properties of rhamnosidase and glucosidase fractions from a fungal flavonoid glycosidase preparation, “Naringinase C-100”. J Food Sci 27:597–601CrossRefGoogle Scholar
  12. Ellenrieder G, Blanco S, Daz M (1998) Hydrolysis of supersaturated naringin solutions by free and immobilized naringinase. Biotechnol Technol 12:63–65CrossRefGoogle Scholar
  13. Elujoba AA, Hardman R (1987) Diosgenin production by acid and enzymatic hydrolysis of fenugreek. Fitoterapia 58:299–303Google Scholar
  14. Erlund I (2004) Review of the flavonoids quercetin, hesperetin, and naringenin. Dietary sources, bioactivities, bioavailability, and epidemiology. Nutr Res 24:851–874CrossRefGoogle Scholar
  15. Esaki S, Ohishi A, Katsumata A, Sugiyama N, Kamiya S (1993) Synthesis of alpha-l-mannopyranosyl-containing disaccharides and phenols as substrates for the alpha-l-mannosidase activity of commercial naringinase. Biosci Biotechnol Biochem 57:2099–2103CrossRefGoogle Scholar
  16. Feng B, Kang L, Ma B, Quan B, Zhou W, Wang Y, Zhao Y, Liu Y, Wang S (2007) The substrate specificity of a glucoamylase with steroidal saponin-rhamnosidase activity from Curvularia lunata. Tetrahedron 63:6796–6812CrossRefGoogle Scholar
  17. Ferreira L, Afonso C, Vila-Real H, Alfaia A, Ribeiro MHL (2008) Debittering of grapefruit juice with naringinase. Food Technol Biotechnol 46:144–148Google Scholar
  18. Fukumoto J, Okada S (1973) Naringinase production by fermentation. Jpn Patent 7(306):554Google Scholar
  19. Gorinstein S, Leontowicz H, Leontowicz M, Krzeminski R, Gralak M, Delgado-Licon E, Ayala ALM, Katrich E, Trakhtenberg SJ (2005) Comparison of the contents of the main antioxidant compounds and the antioxidant activity of white grapefruit and his new hybrid. Agr Food Chem 53:3223–3228CrossRefGoogle Scholar
  20. Habelt K, Pittner F (1983) A rapid method for the determination of naringin, prunin, and naringenin applied to the assay of naringinase. Anal Biochem 134:393–397CrossRefGoogle Scholar
  21. Hall DH (1938) A new enzyme of the glycosidase type. Chem Ind 57:473CrossRefGoogle Scholar
  22. Hoechst (1994) Alpha-rhamnosidase production by Penicillium sp., and purification and characterization. German Patent EP 599, 159Google Scholar
  23. Ito T, Takiguchi Y (1970) Naringinase production by Cochiobolus miyabeanus. Jpn Patent 7(014):875Google Scholar
  24. Jansz ER, Nikawela JK, Gooneratne J, Theivendirarajah K (1994) Studies on the bitter principle and debittering of Palmyrah fruit pulp. J Sci Food Agr 65:185–189CrossRefGoogle Scholar
  25. Jimeno A, Manjon A, Canovas M, Iborra JL (1987) Use of naringinase immobilized on glycophase-coated porous glass for fruit juice debittering. Process Biochem 22:13–16Google Scholar
  26. Kishi K (1955) Production of naringinase from Aspergillus niger. Kagaku to Kogyo. Chem Ind Jpn 29:140Google Scholar
  27. Ko S-R, Choi K-J, Uchida K, Suzuki Y (2003a) Enzymatic preparation of ginsenosides Rg2, Rh1, and F1 from protopanaxatriol-type ginseng saponin mixture. Planta Med 69:285–286CrossRefGoogle Scholar
  28. Ko SR, Choi KJ, Suzuki K, Suzuki Y (2003b) Enzymatic preparation of ginsenosides Rg2, Rh1, and F1. Chem Pharm Bull 51:404–408CrossRefGoogle Scholar
  29. Magario I, Ma X, Neumann A, Syldatk C, Hausmann R (2008) Non-porous magnetic micro-particles: comparison to porous enzyme carriers for a diffusion rate-controlled enzymatic conversion. J Biotechnol 134:72–78CrossRefGoogle Scholar
  30. Magario I, Neumann A, Oliveros E, Syldatk C (2009a) Deactivation kinetics and response surface analysis of the stability of alpha-l-rhamnosidase from Penicillium decumbens. Appl Biochem Biotechnol 152:29–41CrossRefGoogle Scholar
  31. Magario I, Vielhauer O, Neumann A, Hausmann R, Syldatk C (2009b) Kinetic analysis and modeling of the liquid-liquid conversion of emulsified di-rhamnolipids by Naringinase from Penicillium decumbens. Biotechnol Bioeng 102:9–19CrossRefGoogle Scholar
  32. Manjon A, Bastida J, Romero C, Jimeno A, Iborra JL (1985) Immobilization of naringinase on glycophase-coated porous glass. Biotechnol Lett 7:477–482CrossRefGoogle Scholar
  33. Manjon A, Iborra JL, Gómez JL, Gómez E, Bastida J, Bódalo A (1987) Evaluation of the effectiveness factor along immobilized enzyme fixed-bed reactors: design of a reactor with naringinase covalently immobilized into glycophase-coated porous glass. Biotechnol Bioeng 30:491–497CrossRefGoogle Scholar
  34. Manzanares P, Orejas M, Gil JV, Graaff LH, Visser J, Ramon D (2003) Construction of a genetically modified wine yeast strain expressing the Aspergillus aculeatus rhaA gene, encoding an α-l-rhamnosidase of enological interest. Appl Environ Microbiol 69:7558–7562CrossRefGoogle Scholar
  35. Marques J, Vila-Real HJ, Alfaia AJ, Ribeiro MHL (2007) Modelling of the high pressure–temperature effects on naringin hydrolysis based on response surface methodology. Food Chem 105:504–510CrossRefGoogle Scholar
  36. Mateles RI, Perlman D, Humphery AE, Deindorfer FH (1965) Fermentation review. Biotechnol Bioeng 7:54–758CrossRefGoogle Scholar
  37. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428CrossRefGoogle Scholar
  38. Mishra P, Kar R (2003) Treatment of grapefruit juice for bitterness removal by amberlite IR 120 and amberlite IR 400 and alginate entrapped naringinase enzyme. J Food Sci 68:1229–1233CrossRefGoogle Scholar
  39. Nobile MAD, Piergiovanni L, Buonocore GG, Fava P, Puglisi ML, Nicolais L (2003) Naringinase immobilization in polymeric films intended for food packaging applications. J Food Sci 68:2046–2049CrossRefGoogle Scholar
  40. Nomura D (1965) Studies on the naringinase produced by Coniothyrium diplodiella I. The properties of naringinase and the removal of co-existing pectinase from the enzyme preparation. Enzymologia 29:272–282Google Scholar
  41. Norouzian D, Hosseinzadeh A, Inanlou DN, Moazami N (1999) Various techniques used to immobilize naringinase produced by Penicillium decumbens PTCC 5248. World J Microbiol Biotechnol 15:501–502CrossRefGoogle Scholar
  42. Nunes MA, Vila-Real H, Fernandes PC, Ribeiro MHL (2010) Immobilization of naringinase in PVA-alginate matrix using an innovative technique. Appl Biochem Biotechnol 160:2129–2147CrossRefGoogle Scholar
  43. Okada S, Kishi K, Higashihara M, Fukumoto J (1963) Studies on the purification properties of naringinase from Aspergillus niger. J Agric Chem Soc 37:142Google Scholar
  44. Olsons AC, Gray GM, Guadagni DG (1979) Naringin bitterness of grapefruit juice debittered with naringinase immobilized in a hollow fiber. J Food Sci 44:1358–1361CrossRefGoogle Scholar
  45. Orejas M, Ibanez E, Ramon D (1999) The filamentous fungus Aspergillus nidulans produces an α-l-rhamnosidase of potential oenological interest. Lett Appl Microbiol 28:383–388CrossRefGoogle Scholar
  46. Pedro HAL, Alfaia AJ, Marques J, Vila-Real HJ, Calado A, Ribeiro MHL (2007) Design of an immobilized enzyme system for naringin hydrolysis at high-pressure. Enz Microb Technol 40:442–446CrossRefGoogle Scholar
  47. Prakash S, Singhal RS, Kulkarni PR (2002) Enzymatic debittering of Indian grapefruit (Citrus paradasis) juice. J Sci Food Agric 82:394–397CrossRefGoogle Scholar
  48. Puri M, Banerjee U (2000) Production, purification, and characterization of the debittering enzyme naringinase. Biotechnol Adv 18:207–217CrossRefGoogle Scholar
  49. Puri M, Kalra S (2005) Purification and characterization of naringinase from a newly isolated strain of Aspergillus niger 1344 for the transformation of flavonoids. World J Microbiol Biotechnol 21:753–758CrossRefGoogle Scholar
  50. Puri M, Marwaha SS, Kothari RM, Kennedy JF (1996) Studies on the applicability of alginate entrapped naringinase for the debittering of Kinnow juice. Enz Microb Technol 18:281–285CrossRefGoogle Scholar
  51. Puri M, Seth M, Marwaha SS, Kothari RM (2001) Debittering of Kinnow juice by covalent bound naringinase on hen egg white. Food Biotechnol 15:13–23CrossRefGoogle Scholar
  52. Puri M, Kaur H, Kennedy JF (2005) Covalent immobilization of naringinase for the transformation of a flavonoid. J Chem Technol Biotechnol 80:1160–1165CrossRefGoogle Scholar
  53. Puri M, Kaur A, Singh RS, Singh A (2009) Response surface optimization of medium components for naringinase production from Staphylococcus xylosus MAK2. Appl Biochem Biotechnol 162:181–191CrossRefGoogle Scholar
  54. Puri M, Kaur A, Barrow CJ, Singh RS (2010) Citrus peel influences the production of an extracellular naringinase by Staphylococcus xylosus MAK2 in a stirred tank reactor. Appl Microbiol Biotechnol. doi: 10.1007/s0025301028974 Google Scholar
  55. Ribeiro IAC, Ribeiro MHL (2008) Kinetic modelling of naringin hydrolysis using a bitter sweet alfa-rhamnopyranosidase immobilized in k-carrageenan. J Mol Catal B Enz 51:10–18CrossRefGoogle Scholar
  56. Ribeiro IA, Rocha J, Sepodes B, Mota-Filipe H, Ribeiro MHL (2008) Effect of naringin enzymatic hydrolysis towards naringenin on the anti-inflammatory activity of both compounds. J Mol Catal BEnz 52–53:13–18CrossRefGoogle Scholar
  57. Ribeiro MHL, Afonso C, Vila-Real HJ, Alfaia AJ, Ferreira L (2010) Contribution of response surface methodology to the modeling of naringin hydrolysis by naringinase Ca- alginate beads under high pressure. LWT Food Sci Technol 43:482–487CrossRefGoogle Scholar
  58. Roitner M, Schalkhammer Th, Pittner F (1984) Preparation of prunin with the help of immobilized naringinase pretreated with alkaline buffer. App Biochem Biotech 9:483–488CrossRefGoogle Scholar
  59. Saerens K, Bogaert IV, Soetaert W, Vandamme E (2009) Production of glucolipids and specialty fatty acids from sophorolipids by Penicillium decumbens naringinase: optimization and kinetics. Biotechn J 4:517–524CrossRefGoogle Scholar
  60. Sankyo (1988) Preparation of antibiotic chloropolysporin-C. Jpn Patent 63(146):797Google Scholar
  61. Şekeroğlu G, Fadıloğlu S, Göğüş F (2006) Immobilization and characterization of naringinase for the hydrolysis of naringin. Eur Food Res Technol 224:55–60CrossRefGoogle Scholar
  62. Shanmugam V, Yadav KDS (1995) Extracellular production of alpha-rhamnosidase by Rhizopus nigricans. Ind J Exp Biol 33:705–707Google Scholar
  63. Soares N, Hotchkiss F (1998) Naringinase immobilization in packaging films for reducing naringin concentration in grapefruit. J Food Sci 63:61–65CrossRefGoogle Scholar
  64. Soria F, Ellenrieder G, Grasselli M, Navarro del Cañizo AA, Cascone O (2004) Fractionation of the naringinase complex from Aspergillus terreus by dye affinity chromatography. Biotechnol Lett 26:1265–1268CrossRefGoogle Scholar
  65. Spagma G, Barbagallo RN, Martino A, Pifferi PG (2000) A simple method of purifying glycosidase: α-l-rhamnopyranosidases from Aspergillus niger to increase the aroma of Moscato wine. Enzyme Microb Technol 27:522–30CrossRefGoogle Scholar
  66. Suzuki H (1962) Hydrolysis of flavonoid glycosides by enzymes (Rhamnodiastase) from Rhamnus and other sources. Arch Biochem Biophys 99:476–483CrossRefGoogle Scholar
  67. Thammawat K, Pongtanya P, Juntharasri V, Wongvithoonyaporn P (2008) Isolation, preliminary enzyme characterization and optimization of culture parameters for production of naringinase isolated from Aspergillus niger BCC 25166. Kasetsart J Nat Sci 42:61–72Google Scholar
  68. Thomas DW, Smythe CV, Labbee MD (1958) Enzymatic hydrolysis of naringin, the bitter principle of grapefruit. Food Res 23:591–598Google Scholar
  69. Ting SV (1958) Enzymatic hydrolysis of naringin in grapefruit. J Agric Food Chem 6:546–549CrossRefGoogle Scholar
  70. Tsen H-Y, Tsai S-Y (1988) Comparison of the kinetics and factors affecting the stabilities of chitin-immobilized naringinases from two fungal sources. J Ferment Technol 66:193–198CrossRefGoogle Scholar
  71. Tsen H-Y, Tsai S-Y, Gee-Kaite Yu (1989) Fiber entrapment of naringinase from Penicillium sp. and application to fruit juice debittering. J Ferm Bioeng 67:186–189CrossRefGoogle Scholar
  72. Vila-Real HJ, Alfaia AJ, Calado AT, Ribeiro MHL (2007) High pressure–temperature effects on enzymatic activity: naringin bioconversion. Food Chem 102:565–570CrossRefGoogle Scholar
  73. Vila-Real H, Alfaia AJ, Emilia Rosa M, Calado ART, Ribeiro MHL (2010a) An innovative sol–gel naringinase bioencapsulation process for glycosides hydrolysis. Process Biochem 45:841–850CrossRefGoogle Scholar
  74. Vila-Real H, Alfaia AJ, Rosa J, Góis P, Rosa ME, Calado ART, Ribeiro MHL (2010b) α-Rhamnosidase and β-glucosidase expressed by naringinase immobilized on new ionic liquid sol–gel matrices: activity and stability studies. J Biotechnol. doi: 10.1016/j.jbiotec.2010.08.005 Google Scholar
  75. Vila-Real H, Alfaia AJ, Calado ART, Ribeiro MHL (2010c) Improvement of activity and stability of soluble and sol–gel immobilized naringinase in co-solvent systems. J Mol Catal B Enz 65:91–101CrossRefGoogle Scholar
  76. Yanai T, Sato M (2000) Purification and characterization of α-l-rhamnosidases from Pichia angusta X349. Biosci Biotechnol Biochem 64(10):2179–85CrossRefGoogle Scholar
  77. Young NM, Johnston RAZ, Richards JC (1989) Purification of the α-l-rhamnosidase of Penicillium decumbens and characterisation of two glycopeptide components. Carbohydr Res 191:53–62CrossRefGoogle Scholar
  78. Zverlov VV, Hertel C, Bronnenmeier K, Hroch A, Kellermann J, Schwarz WH (2000) The thermostable alpha-l-rhamnosidase RamA of Clostridium stercorarium: biochemical characterization and primary structure of a bacterial alpha-l-rhamnoside hydrolase, a new type of inverting glycoside hydrolase. Mol Microb 35:173–179CrossRefGoogle Scholar

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© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Research Institute for Medicines and Pharmaceutical Sciences (i.Med-UL), Faculty of PharmacyUniversity of LisbonLisbonPortugal

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