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Production, purification, and characterization of the debittering enzyme naringinase


Biotechnology Advances 18 (2000) 207–217

Research review paper

Production, purification, and characterization of the debittering enzyme naringinase
Munish Puri, Uttam Chand Banerjee*
Biochemical Engineering Research and Process Development Centre, Institute of Microbial Technology, Sector 39-A, Chandigarh 160036, India

Abstract This review discusses the debittering enzyme naringinase and its essential role in the commercial processing of citrus fruit juice. Applications of this enzyme in other areas are identified. Characterization of the enzyme is detailed and its immobilized preparations are discussed. Production of microbial naringinase by fermentation is described. ? 2000 Elsevier Science Inc. All rights reserved.
Keywords: Naringinase; Naringin; Debittering of fruit juice

1. Introduction The fruit juice industry has to deal with raw material that varies greatly in its many quality attributes. The industry aims to process fruit at the lowest possible cost while maintaining organoleptic quality and stability of the finished product. The enzymes naringinases are used in orange and grapefruit processing to improve pulp washing, increase the recovery yield of essential oils, and to debitter and clarify the juice (Grassin and Fauquembergue, 1996). The most bitter compounds present in citrus juices are naringin, limonin, and neohesperidin (Kefford, 1959; Marwaha et al., 1994). These bitter compounds are found in all parts of grapefruit and sour oranges. Naringin is the major component in grapefruit and it is by far the most bitter. Its taste threshold in water is approximately 20 ppm, but 1.5 ppm levels may be detected. Naringin is abundant in immature fruit but its concentration decreases as fruit ripens (Yusof et al., 1990). All processed grapefruit juice contains naringin above 50 ppm level; limonin present in the processed juice acts synergistically with naringin to cause bitterness. The presence of bitterness has been a major limitation in the commercial acceptance of juice. The naringin level can be reduced by technologies such as adsorptive debittering (Grif* Corresponding author. Tel. 91-172-690-908; fax: 91-172-690-585. E-mail address: uttam@koel.imtech.ernet.in (U.C. Banerjee) 0734-9750/00/$ – see front matter ? 2000 Elsevier Science Inc. All rights reserved. PII: S0 7 3 4 - 9 7 5 0 ( 0 0 ) 0 0 0 3 4 -3

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fith, 1969; Johnson and Chandler, 1988), chemical methods (Kimball, 1987; Pritchet, 1957), treatment with polystyrene divinyl benzene styrene (DVB) resins (Kimball, 1991; Puri, 1984), and -cyclodextrin treatment (Shaw and Wilson, 1983; Wagner et al., 1988). These technologies have the following inherent limitations: (1) the juice must be previously deoiled, dewaxed, depulped, and then reblended with the clarified debittered juice; (2) adsorption columns are usually regenerated with dilute alkali solution and this may affect organoleptic properties and final quality of juice; (3) the methods may alter the composition of the juice either through chemical reaction(s) or removal of nutrients, flavor, color, etc.; (4) the methods are nonspecific, inherently inefficient, and they introduce batch-to-batch variation because of non-monitorable changes; (5) the methods of extraction and finishing affect the yield, quality, and characteristics of citrus juice produced. Methods such as the acid hydrolysis of naringin produce not only rhamnose and glucose but also the bitter aglycon and naringenin. Therefore, acid hydrolysis is not suited to commercial processing. Similarly, under selected conditions of pH and temperature, activated charcoal may almost completely remove naringin from a solution; however, many of the desirable flavoring components are simultaneously removed. A radial flow chromatographic process has been described for debittering of fruit juice (Coghlan, 1997). In comparison with conventional chromatography, the radial flow systems are faster and operate at lower pressures (Chisti, 1998). The chromatographic resin in this patented process captures naringin as the juice washes through. The processed juice has the requisite body and flavor, but not the bitter taste (Coghlan, 1997). Whether this process gains commercial acceptance remains to be seen. Because of the various drawbacks, the capabilities of non-enzymatic debittering technologies are limited. A suitable debittering procedure is stepwise hydrolysis of naringin by naringinase (Habelt and Pittner, 1983; Ting, 1958). The naringinase activity corresponds to rhamnosidase and glucosidase activities, working sequentially. The use of enzymes for debittering is increasing rapidly. Enzymatic processes also minimize pollution during processing. An immobilized enzyme system consisting of amylase and polygalacturonase co-immobilized with naringinase has been used in processing of vegetable and fruit juice. This co-immobilized system yields more rapid starch and protein degradation and juice clarification than separately immobilized preparations of amylase and pectinase (Magindag, 1989). Despite many attractive features, the enzymatic debittering of citrus fruit juice is limited at present because of the following factors: (1) procurement of enzymes is expensive for processing large volumes of juice; (2) the limonin content is not at all affected by naringinase treatment; (3) the availability of neutral ion exchange resin technology for debittering and deacidifying the grapefruit juice; and (4) the inability of the available naringinase preparations to completely hydrolyze naringin. This review discusses some of the options available for enhanced utilization of naringinase, its production and characterization.

2. Applications of naringinase Naringinase has other important applications and potential applications besides debittering of fruit juice. Some of the applications are discussed.

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2.1. Preparation of the antibiotic chloropolysporin C The deglycosylation of novel glycopeptide antibiotic, chloropolysporin from Faenia interjecta, was achieved successfully with rhamnosidase activity of naringinase. Chloropolysporins A, B, and C could be enzymatically converted to deglycosylated derivatives (Sankyo, 1988). A combination of chloropolysporin C and -lactam antibiotics is synergistically effective against methicillin resistant strains of Staphylococcus. Chloropolysporins have strong activity against gram-positive bacteria including methicillin resistant Staphylococcus aureus and these antibiotics also inhibit the anaerobic gram-positive Enterobacteria. Generally, chloropolysporin C is the most active component of the chloropolysporin complex and its activity is slightly stronger than that of -avoparcins. 2.2. Preparation of rhamnose Naringinases ( -L-rhamnosidases) hydrolyze naringin to produce L-rhamnose. The enzyme mixture used is a partially purified preparation with a high rhamnosidase activity and low glucosidase activity (Daniels et al., 1990). Rhamnose is a chiral intermediate in organic synthesis and it is used as a pharmaceutical and a plant protective agent. 2.3. Preparation of prunin The flavonoid prunin may be produced from naringin using immobilized naringinase pretreated with an alkaline buffer. The product is obtained in high yield. Prunin has variable antiviral activity against DNA RNA viruses. The flavonoid possesses anti-inflammatory activity and may be used as sweetening agent for diabetics (Roitner et al., 1984a). Many naturally occurring flavonoids are known to possess a variable spectrum of antiviral activity against certain RNA (e.g. RSV, Pf-3, polio) and DNA (e.g. HSV-1) viruses. The effects of flavonoids such as naringin, hesperitin, and prunin on the infectivity and replication of herpes simplex virus type 1 (HSV-1), polio-virus type-1, parainfluenza virus type 3 (Pf-3), and the respiratory syncytial virus (RSV) have been studied in vitro in cell culture monolayers (Kaul et al., 1985). The natural flavonone glycoside of naringenin has also been reported to prevent gastric mucosal ulceration in animal models. Naringin itself is reported to have gastroprotective effect in ethanol-induced gastric injury. 2.4. The rhamnosidase activity of naringinase The rhamnosidase activity of naringinase in combination with -glucosidase and arabinosidase is considered suitable for aroma enhancement in wine making. In one study, the enzymes were immobilized to a solid carrier with the aim of developing a continuous process for wine aroma enhancement (Caldini et al., 1994). 2.5. Steroid transformation The fenugreek seeds (Trigonella foenumgraecum) upon enzymatic hydrolysis by naringinase produces sapogenins and diosgenin (a precursor of clinically useful steroid drugs). In this process, the immobilized naringinase and pectinase could be reused twice without loss of diosgenin yield (Elujoba and Hardman, 1987).

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3. Mechanism of naringinase action The available information throws little light on the molecular mechanism of enzyme action. Naringinase is an enzyme complex consisting of -rhamnosidase (EC 3.2.1.40) and flavonoid- -glucosidase (EC 3.2.1.21). In typical processing, naringinase converts naringin to naringenin in a two-step process (Fig. 1). The substrate naringin, 4 -5,7 -trihydroxyflavonone-7-rhamnoglucoside, is hydrolyzed by the rhamnosidase component to produce prunin (4 -5,7 -trihydroxyflavonone-7-glucoside), which is then converted by the flavonoid -glucosidase to naringenin (4 -5,7 -trihydronyflavonone) (Chandler and Nicol, 1975; Habelt and Pittner, 1980). Naringenin is only one-third as bitter as naringin; however, prunin is less bitter than naringenin and only the first hydrolyzing activity of naringinase is in fact essential for bitterness removal. 4. Production of naringinase Historically, naringinase has been isolated from plant sources such as celery seeds (Hall, 1938) and grapefruit leaves (Hall, 1938, Thomas et al., 1958; Ting, 1958); however, for reasons of availability, only processes based on microbial naringinases are practicable. There are only a few reports on the microbial production of naringinase. In many instances the production methods are patented and only sketchily reported. Naringinase is produced by many microorganisms, as noted in Table 1. Microbial naringinase may be produced both by submerged culture and solid-state fermentation processing, as discussed. 4.1. Submerged culture A large number of microorganisms have been screened for their naringinase producing ability (Thomas et al., 1958). Thomas et al. (1958) used crude culture extracts of microorganisms to obtain a naringinase preparation C that had an optimum pH of 5–6 and could be held at 60 C for 4 h with only 16% loss in activity. Neither the culture extracts stored at 5 C, nor the enzyme stored at room temperature showed any loss in activity during one year. The isolated enzyme was partially purified by alcohol precipitation of the culture extracts. One study focused on molds, explored 96 strains and established Aspergillus niger as the best producer of naringinase (Kishi, 1955). Later, Okada et al. (1963) purified naringinase produced by this strain and reported on the properties of the enzyme. Using these studies as the basis, the Tanabe Pharmaceuticals Company established a process for producing a naringinase preparation that was marketed as ‘Kumitanase.’ In 1960, Smythe and Thomas filed a U.S. patent that described the production of enzyme at approximately 100 U mL (Smythe and Thomas 1960). Future studies focused on increasing the productivity and the activity of the enzyme preparations.

Fig. 1. Stepwise degradation of naringin by the action of naringinase.

M. Puri, U.C. Banerjee / Biotechnology Advances 18 (2000) 207–217 Table 1 Microorganisms used for the production of naringinase Microorganism Aspergillus niger, Aspergillus oryzae, A. usamii Cochiobolus miyabeanus Coniothyrium diplodiella Penicilium decumbens Phanopsis citri Rhizotonia solani Rhizopus nigricans References

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Bram and Solomons, 1965; Kishi, 1955 Ito and Takiguchi, 1970 Nomura, 1965 Fukumoto and Okado, 1973 Ito and Takiguchi, 1970 Ito and Takiguchi, 1970 Shanmugam and Yadav, 1995

Bram and Solomons (1965) observed that the production of naringinase in A. niger was repressed by glucose. The production of enzyme decreased below pH 4 and was stimulated in the presence of the substrate (naringin). A variety of complex media were examined for enzyme production in shake flasks. The cornsteep liquor–yeast extract medium provided the best enzyme titers. The stepwise addition of small amounts of naringin was more effective in stimulating enzyme production than a higher concentration of the substrate added at the beginning of the fermentation. A standard spore suspension of 105 spores mL was used for the initiation of the fermentation. Fermentations were carried in a 10-L bioreactor, fully baffled, with disc turbine impellers, and automatic control of temperature and foam. Other work revealed enhanced naringinase production if the culture medium contained rhamnose or plant meal that had rhamnose (Mateles et al., 1965). In addition to glucose, lactate and citrate are also known to suppress the production of naringinase by A. niger NRRL 72-4. Sucrose and starch similarly suppress enzyme production, although these components support excellent growth. The inclusion of starch as whole corn meal completely represses enzyme synthesis. The production of naringinase is inducer-dependent and continuous addition or stepwise addition of an inducer increases naringinase production. Whereas, replacement of the inducer with other carbon sources supports the growth of the organism but no enzyme is produced. A phytopathogenic microorganism Coniothyrium diplodiella was used by the Sankyo Company of Japan to produce a pectic enzyme preparation that had naringinase activity. This preparation was marketed as ‘Sankyo naringinase’ and reported to have a high potency (Takiguchi, 1962). Ito and Takiguchi (1970) registered a patent for producing naringinase from Phanopsis citri, Rhizotonia solani, and Cochiobolus miyabeanus. The 5-day fermentations used media based on soybean meal and were carried out at 25 C. Fukumoto and Okada (1973) reported naringinase production using Penicillium sp. The fermentation medium comprised of soybean residue and cornsteep liquor. The process depended on the inducer hesperidin. The batch fermentation lasted 5 days at 30 C. For nearly two decades since these pioneering studies, no significant developments appear to have been reported on submerged culture production of naringinase. Then, in 1994, a patent was registered by Hoechst (1994) on the -rhamnosidase production, purification and characterization from Penicillium sp. The enzyme, produced by fermentation of Penicillium DSM 6825, was concentrated and purified from the culture broth. The enzyme had a molecular weight of 60–100 kDa with an specified N-terminal sequence and it catalyzed cleavage of the bond between terminal rhamnose and the aglycon in a rhamnose containing glycoside. The enzyme had an isoelectric pH

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of 5.6–5.8; its pH and temperature optima were 5–5.5 and 50–55 C, respectively. This enzyme had a different specificity than for the commonly known naringinase which preferentially cleaves two monosaccharide residues. The purified enzyme has been used to produce L-rhamnose. Shanmugam and Yadav (1995) demonstrated the extracellular production of L-rhamnosidase from a strain of the fungus Rhizopus nigricans. The sterilized culture medium contained sucrose and rice and was inoculated with spore suspension (106 spores mL). The naringinase activity peak was observed nearly 50 h after inoculation. The pH of the medium declined as mycelia grew. The extracellular culture filtrate containing -rhamnosidase was dialyzed overnight against distilled water at 30 C. The enzyme was shown to obey Michaelis-Menten kinetics when tested with p-nitrophenyl -L-rhamnopyranoside as the substrate. The pH and temperature optima of the enzyme were pH 6.5 and 60–80 C, respectively. In all reported fermentation processes, the naringinase was observed in the extracellular broth. Submerged fermentation appears to dominate in commercial production of naringinase. Submerged fermentations are relatively easily controlled and scaled up. Moreover, yields are generally higher in submerged culture and the risk of contamination is low. 4.2. Solid-state fermentation Compared to studies with submerged culture, solid-state fermentation has been barely investigated for production of naringinase. However, there is substantial scope for this mode of production, as revealed by demonstrated automation capabilities and operating experience with many other large-scale solid-substrate fermentation processes (Chisti, 1999). Production of naringinase by solid-state culture of Coniothrium diplodiella was reported by Nomura (1965). The micro-organism was grown on soybean cake at 23 C and for 8 days. The resulting koji cake was dried, ground, and extracted to obtain a crude preparation of naringinase. The crude enzyme was further purified to remove the contaminating pectinase. The optimum pH and temperature for the naringinase were pH 4.2 and 60–65 C, respectively. The naringinase produced was inhibited considerably by sucrose and fructose, and to a lesser extent by sorbitol.

5. Determination of naringinase activity Only a few assay procedures for naringinase are presently available. These procedures are based on the spectrophotometric determination of flavonones according to the alkaline diethylene glycol method of Davis (1947). The naringin reacts with the reagent to produce a yellow color which is measured at a wavelength of 420 nm. A high performance liquid chromatographic (HPLC) determination has also been used for naringinase activity (Horuichi et al., 1985). The HPLC method determines activity of the enzyme by measuring changes in concentration of -rhamnoside. Romero et al. (1985) used p-nitrophenyl-L-rhamnopyranoside for the measurement of L-rhamnosidase activity of naringinase, by colorimetrically following the appearance of p-nitrophenol. The use of a synthetic substrate did not affect the pH, temperature, and ionic strength optima of the enzyme. Using naringin as substrate, the naringinase activity was determined by HPLC on C18 Bondapak reverse-phase column and detection of absorbance at 280 nm.

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6. Characterization of naringinase Naringinase from various sources has been characterized in some detail. The enzyme isolated from Penicillium sp. is a glycoprotein which is electrophoretically homogenous, as determined by electrofocussing and SDS gel electrophoresis studies (Gabor and Pittner, 1984). The enzyme possesses both -rhamnosidase (EC 3.2.1.40) and -D-glucosidase (EC 3.2.1.24) activities. The pH optima for the two activities were 4.5 and 3.0, respectively (Gabor and Pittner, 1984). Using gel filtration, electrophoresis and immobilization, Schalkhammer and Pittner (1986) determined the holoenzyme and subunit structure for this enzyme. Light scattering and exclusion chromatography indicated a clear dependence of the molecular mass on the pH of buffer. Using data on the kinetic parameters Km (Michaelis constant), Vmax (the maximum rate), and Ki (the inhibition constant) for glucose and rhamnose, Schalkhammer and Pittner (1986) ascertained the existence of two catalytic sites. The chemical modification of both the active site regions supported the existence of an essential tyrosine site, as being necessary for both the activities. Modification of lysine group at one of the sites increased glucosidase activity by up to 5-fold. The commercially available naringinase of Aspergillus niger also has both -rhamnosidase and -D-glucosidase activities (Roitner et al., 1984b). The ratio of these activities varies with the protein concentration and the pH. The rhamnosidase activity is nearly independent of pH in the range from 3 to 7, whereas glucosidase activity shows a distinct optimum that varies between pH 4 and 6, depending on the pretreatment used. The enzyme complex can be separated into various oligomers by gel filtration. The naringinase of A. niger appears to be a single enzyme with two active sites, one for the -L-rhamnosidase activity and the other for -D-glucosidase activity. 7. Immobilized naringinase Many immobilized preparations of naringinase have been successfully used in debittering operations and for producing biochemicals such as rhamnose and prunin. Some of the immobilized preparations are discussed. 7.1. Aspergillus naringinase The naringinase of A. niger immobilized on copolymers of styrene and maleic anhydride has been used to hydrolyze naringin (Goldstein et al., 1971). In another case, naringinase immobilized on a tannin-aminohexyl cellulose showed a half-life of 198 days at 25 C and 88 days at 37 C during debittering of Citrus natsudaidai juice (Ono et al., 1978). In the third case, naringinase immobilized on chitin with glutaraldehyde and sodium borohydride was inactivated during debittering of grapefruit juice (Tsen, 1984). Glucose, fructose, and rhamnose were found to inhibit the enzyme, regardless of whether it was in the native or immobilized state. Both the source of the juice and the immobilization system used may affect the stability of the enzyme preparation. In a hollow-fiber reactor for the hydrolysis of naringin in unclarified grapefruit juice, naringinase could be used for 4 h at 66% efficiency (Olson et al., 1979). In further rigorous testing of this system, high levels (900 g mL) of naringin in grapefruit juice were reduced to acceptably low values (Gray and Olson, 1981). The effects

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of operational parameters (flow rates, membrane surface area, temperature, and enzyme loading) on naringin hydrolysis to prunin and naringenin, were reported (Gray and Olson, 1981). Roitner et al. (1984a) immobilized the solid phase form of naringinase on controlled pore glass in an enzyme reactor for the conversion of naringin to prunin. The process was carried out in 0.1 M glycine NaOH buffer at pH 12; 98% conversion was achieved. The naringenin in the product stream was extracted with CHCl3 and the unreacted substrate in the aqueous phase was returned to the reactor. This was claimed to be the first problem-free process for the production of prunin (Roitner et al., 1984a). 7.2. Penicillium naringinase The naringinase derived from Penicillium sp. has a high activity of -L-rhamnosidase and a low activity of -D-glucosidase. The enzyme immobilized on silicate and polysaccharides by using glutaraldehyde, is known to cleave anthraquinone and steroid glucoside in batch and continuous reactors. Immobilization of naringinase on silicate does not interfere with the analysis of the liberated L-rhamnose (Turecek and Pittner, 1987). Naringinase of Penicillium sp. when entrapped in cellulose triacetate fibers shows higher Km-values than in its soluble form (Tsen et al., 1989). The fiber-entrapped enzyme has been used for hydrolyzing naringin in real and simulated grapefruit juice. This system was also used to simultaneously degrade limonin while hydrolyzing naringin (Tsen and Yu, 1991). When grapefruit juice was debittered with this enzyme column, the sugar components, the total organic acids, and the turbidity levels remained unaltered. Moreover, the column could be regenerated by washing with warm water. In another case, naringinase from Penicillium sp. was covalently linked to glycophase coated controlled pore glass (CPG) and used in debittering of fruit juice (Manjon et al., 1985). The operational stability of this preparation was tested in a packed bed reactor (Jimeno et al., 1987). Theoretical models were used to predict the behavior of the system and good agreement with measured data were obtained (Manjon et al., 1985). 7.3. Other systems According to one source (Magindag, 1989), a system of co-immobilized amylase, polygalacturonase, and naringinase is superior for starch and pectin hydrolysis in fruit and vegetable juices relative to one in which the juice is treated with separately immobilized pectinase and amylase. In this work, the enzymes were co-immobilized by heating the enzymes and the silicate support suspension in water, adding glutaraldehyde, and washing of the resulting solids. Caldini et al. (1994) conducted kinetic and immobilization studies in fungal glucosidases for aroma enhancement in wine. The enzyme preparation of A. niger contained -glucosidase, -arabinosidase, and -rhamnosidase activities in a ratio considered suitable for aroma enhancement in wine making. The three activities were immobilized to a silianized bentonite solid carrier with glutaraldehyde with the aim of developing a continuous process for wine aroma enhancement. Puri et al. (1996) worked on the use of alginate-entrapped naringinase for debittering of kinnow juice. Various alginate matrices were screened for the immobilization and a matrix obtained with 2% sodium alginate was the optimal matrix system. A preparation that contained 30 units of naringinase, hydrolyzed 82% of initial naringin in 3 h. The

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immobilization broadened the pH optimum and improved the flexibility of the process for debittering juice of varying pH values. Immobilization also enhanced the thermal stability of the enzyme. The application of kinetic parameters optimized with pure naringin to kinnow juice resulted in 60% debittering. The use of covalently immobilized naringinase has also been proposed for debittering of kinnow juice (Puri et al., 1997). The wood chip matrix used was inexpensive and required a one-step activation. The covalent binding of the enzyme to wood was through glutaraldehyde. The enzyme did not leach from the support. The system required a contact time of 5 h for debittering of the kinnow juice.

8. Concluding remarks As judged from limited data, naringinase is the subject of much continuing research and it is commercially attractive. It is potentially useful in food, pharmaceutical, and flavoring applications; however, a limited supply of the enzyme restricts wider use. Commercially viable processes are needed for large-scale production of the enzyme by fermentation. Only a few reports address fermentative production of naringinase at large scale and its subsequent use in debittering of fruit juice.

Acknowledgments One of the authors (M.P.) thanks the Department of Biotechnology, Government of India, for financial support. This is IMTECH communication 044 97.

References
Bram B, Solomons GL. Production of the enzyme naringinase by Aspergillus niger. Appl Microbiol 1965;13:842–5. Caldini C, Bonomi F, Pifferi PG, Lanzarini G, Galente YM. Kinetic and immobilization studies on fungal glycosidase for aroma enhancement in wine. Enz Microb Technol 1994;16:286–91. Chandler BV, Nicol KJ. Some relationships of naringin: their importance in orange juice bitterness. CSIRO Food Res Quart 1975;35:79–88. Chisti Y. Strategies in downstream processing. In: Subramanian G, editor. Bioseparation and Bioprocessing: A Handbook, vol. 2. New York: Wiley-VCH, 1998. pp. 3–30. Chisti Y. Solid substrate fermentations, enzyme production, food enrichment. In: Flickinger MC, Drew SW, editors. Encyclopedia of Bioprocess Technology: Fermentation, Biocatalysis, and Bioseparation, vol. 5. New York: Wiley, 1999. pp. 2446–62. Coghlan A. Squeezing bitterness out of grapefruit. Nature Biotechnol 1997;15:202. Daniels L, Linhardt RJ, Bryan BA, Mayerl F, Pickenhagen M. Methods for producing rhamnose. US Patent 4,933,281. 1990. Davis DW. Determination of flavonones in citrus juice. Anal Chem 1947;19:46–8. Elujoba AA, Hardman R. Diosgenin production by acid and enzymatic hydrolysis of fenugreek. Fitoterapia 1987;58:299–303. Fukumoto J, Okada S. Naringinase production by fermentation. Japanese Patent 7,306,554. 1973. Gabor F, Pittner F. Characterization of naringinase from Penicillium sp. Hoppe Seyler-Z Physiol Chem 1984;365:914–6.

216

M. Puri, U.C. Banerjee / Biotechnology Advances 18 (2000) 207–217

Goldstein L, Lifshitz A, Skolovsky M. Water insoluble derivatives of naringinase. Int J Biochem 1971;10:448–9. Grassin C, Fauquembergue P. Fruit juices. In: Godfrey T, West S, editors. Industrial Enzymology, 2nd edition. New York: Macmillan Press, 1996, pp. 225–64. Gray GM, Olson AC. Hydrolysis of high levels of naringin in grapefruit juice using a hollow fibre naringinase reactor. J Agric Food Chem 1981;29:1298–301. Griffith FP. Process for reactivating polyamides resins used in debittering citrus juice. US Patent 3,463,763. 1969. Habelt K, Pittner F. A rapid method for the determination of naringin, prunin, and naringin applied to the assay of naringinase. Anal Biochem 1983;134:393–7. Hall DH. A new enzyme of the glycosidase type. Chem Ind 1938;57:473. Hoechst. Alpha-rhamnosidase production by Penicillium sp., and purification and characterization. German Patent EP 599, 159. 1994. Horuichi S, Yamamoto H, Asakoshi T, Tanaka T. High pressure liquid chromatographic determination of naringinase activity. J Jpn Soc Food Sci Technol 1985;32(8):582–5. Ito T, Takiguchi Y. Naringinase production by Cochiobolus miyabeanus. Japanese Patent 7,014,875. 1970. Jimeno A, Manjon A, Canvos M, Iborra JL. Use of naringinase immobilized on glycophase coated porous glass for fruit juice debittering. Process Biochem 1987;22:13–7. Johnson RL, Chandler BV. Adsorptive removal of bitter principles and titrable acid from citrus juice. Food Technol 1988;45:130–7. Kaul TN, Middleton E, Ogra PL. Antiviral effect of flavonoids on human viruses. J Med Virol 1985;15:71–9. Kefford JF. The chemical constituents of citrus fruits. Adv Food Res 1959;9:285. Kimball DA. Debittering of citrus juices using supercritical carbon dioxide. J Food Sci 1987;52:481–2. Kimball DA. Citrus Processing: Quality Control Technology. New York: Van Nostrand Reinhold, 1991. Kishi K. Production of naringinase from Aspergillus niger. Kagaku to Kogyo (Chemistry and Industry, Japan) 1955;29:140. Magindag. Amylase, polygalacturonase and naringinase co-immobilization. German Patent EP 298,954. 1989. Manjon A, Bastida J, Romero C, Jimeno A, Iborra JL. Immobilization of naringinase on glycophase coated porous glass beads. Biotechnol Let 1985;7:487–92. Marwaha SS, Puri M, Bhular M, Kothari RM. Optimization of parameters for the hydrolysis of limonin for debittering of kinnow mandarin juice by Rhodococcus fascians. Enz Microb Technol 1994;16:723–5. Mateles RI, Perlman D, Humphery AE, Deindorfer FH. Fermentation review. Biotechnol Bioeng 1965;7:54–8. Nomura D. 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 1965;29:272–82. Okada S, Kishi K, Higashihara M, Fukumoto J. Studies on the purification properties of naringinase from Aspergillus niger. J Agric Chem Soc (Japan) 1963;37:142. Olson AC, Gray GM, Guadagni DG. Naringinase bitterness of grapefruit juice debittered with naringinase immobilized in a hollow fibre. J Food Sci 1979;44:1358–60. Ono M, Tosa T, Chibata I. Preparation and properties of immobilized naringinase using tannin-aminohexylcellulose. Agric Biol Chem 1978;42:1847–53. Pritchet DE. Extraction of the bitter principle from navel orange juice. US Patent 2,816,033. 1957. Puri A. Preparation and properties of citrus juices, concentrates and dried powders which are reduced in bitterness. US Patent 4,439,458. 1984. Puri M, Marwaha SS, Kothari RM. Studies on the applicability of alginate entrapped naringinase for the debittering of kinnow fruit juice. Enz Microb Technol 1996;18:281–5. Puri M, Dharni HH, Marwaha SS. The applicability of covalently immobilized naringinase for the debittering of kinnow fruit juice. In International conference on Bioencapsulation VI, Barcelona, Spain, 30 Aug–Sep 1, 1997. Roitner M, Schalkhammer T, Pittner F. Preparation of prunin with the help of immobilized naringinase pretreated with alkaline buffer. Appl Biochem Biotechnol 1984a;9:483–8. Roitner M, Schalkhammer T, Pittner F. Characterization of naringinase from Aspergillus niger. Monatsh Cheme 1984b;115:1255–67. Romero C, Manjon A, Bastida J, Iborra JL. A method for assaying rhamnosidase activity of naringinase. Anal Biochem 1985;149:566–71.

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Sankyo. Preparation of antibiotic chloropolysporin-C. Japanese Patent 63,146,797. 1988. Schalkhammer T, Pittner F. Characterization of rhamno-glucosidase from Penicillium sp. Biol Chem Hoppe-Seyler 1986;367, 364 (conference abstract). Shanmugam V, Yadav KDS. Extracellular production of alpha-rhamnosidase by Rhizopus nigricans. Ind J Exp Biol 1995;33:705–7. Shaw PE, Wilson CW. Debittering citrus juices with -cyclodextrin polymer. J Food Sci 1983;48:646–7. Smythe CW, Thomas DW. Conversion of flavonoid glycosides. US Patent 2,950,974. 1960. Takiguchi Y. Annual Reports of Takamine Institute, Tokyo. Agric Soc Jpn 1962;14:101. Thomas DW, Smythe CV, Labbee MD. Enzymatic hydrolysis of naringin, the bitter principle of grapefruit. Food Res 1958;23:591–8. Ting SV. Enzymatic hydrolysis of naringin in grapefruit. J Agric Food Chem 1958;6:546–9. Tsen HY. Factors affecting the inactivation of naringinase immobilized on chitin during debittering from fruit juices. J Ferment Technol 1984;62:263–7. Tsen HY, Tsai SY, Yu GJ. Fibre entrapment of naringinase from Penicillium sp. and application to fruit juice debittering. J Ferment Technol 1989;67:186–9. Tsen HY, Yu GJ. Limonin and naringin removal from grapefruit juice with naringinase entrapped in cellulose triacetate fibres. J Food Sci 1991;56:31–4. Turecek PL, Pittner F. Application of enzyme immobilization to the analysis of naturally occurring compounds as exemplified by alpha-rhamnosides. Sci Pharm 1987;55:275–83. Wagner CJ, Wilson CW, Shaw PE. Reduction of grapefruit bitter components in a fluidized -cyclodextrin polymer bed. J Food Sci 1988;53:516–8. Yusof S, Gazali HM, King GS. Naringin content in local citrus fruit. Food Chem 1990;37:113–21.


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