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Pineapple Juice and Its Fractions in Enzymatic Browning
Inhibition of Banana [Musa (AAA Group) Gros Michel]
Chitsuda Chaisakdanugull, Chockchai Theerakulkait, and Ronald E. Wrolstad
J. Agric. Food Chem., 2007, 55 (10), 4252-4257• DOI: 10.1021/jf0705724 • Publication Date (Web): 18 April 2007
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4-dodecyl-.00 © 2007 American Chemical Society Published on Web 04/18/2007 and Bangkok 10900. 4252−4257 Pineapple Juice and Its Fractions in Enzymatic Browning Inhibition of Banana [Musa (AAA Group) Gros Michel] CHITSUDA CHAISAKDANUGULL. WROLSTAD‡ Department of Food Science and Technology. Further identification of the DE fraction by fractionation with ion exchange chromatography and confirmation using model systems indicated that malic acid and citric acid play an important role in the enzymatic browning inhibition of banana PPO. additional compounds. These two thiol compounds block the o-quinone by forming Fax: +66-2-562-5021. McEvily (8) reported that ficin- effectiveness as a browning inhibitor. Thailand. grape juice. Oszmianski and Lee (5) reported that honey can inhibit compounds were identified as cysteine and a dipeptide of cysteine-glutamic acid. reducing agents. Various chemicals such as 4-hexyl-.2 µM.† AND RONALD E. especially bananas. However. pineapple juice. 55. Oregon State University. however.5. Food Chem. including I50 values of 0. After storage of banana slices at 15 °C for 3 days. The brown color is developed of honey appeared to be a small peptide with an approximate due to the enzymatic oxidation of phenols to quinones by molecular mass of 600 Da. In contrast. sulfites (4). E-mail: fagicct@ku. Monsalve- PPO inhibitors. o-diphenol oxidase. The agents. and Department of Food Science and Technology. 0. KEYWORDS: Polyphenol oxidase. noncompetitive inhibitors of with other substances such as phenolic compounds and amino mushroom PPO (6). however. * To whom correspondence should be addressed. The compound responsible for this inhibitory effect their tissues are cut or bruised. and model Many types of fruits. (7) investigated the effect of proteolytic enzymes also known as tyrosinase. browning inhibition. The DE fraction also showed more inhibitory effect than 8 mM ascorbic acid in enzymatic browning inhibition of banana puree during storage at 5 °C for 24 h. Corvallis. PPO (1. Faculty of Agro-industry. The component that was are known to induce adverse allergenic effects in certain responsible for browning inhibition was 4-substituted resorcinol sensitive individuals such as asthmatics (3). Tel: +66-2-562-5032. commercially available as γ-Glu-Cys. enzymatic browning. and 4-cyclohexylresorcinol with are known to be able to reduce enzymatic browning.1021/jf0705724 CCC: $37. these quinones condense and react nonenzymatically acids. 2007. brown rapidly when systems. and chelating agents. banana INTRODUCTION enzymatic browning in apple slices.*. Subsequent research has demonstrated that dangers of synthetic food additives. Subse. sulfites browning in apple and shrimp. bromelain was effective only in To prevent browning. the PPO inhibitors in honey were t-cinnamic and p-coumaric quently.† CHOCKCHAI THEERAKULKAIT. Enzymatic browning impairs not only the color of papain were as effective as sulfite in preventing browning in fresh fruits but also the flavor and the nutritional quality (2). Fractionation of pineapple juice by a solid-phase C18 cartridge revealed that the directly eluted fraction (DE fraction) inhibited banana polyphenol oxidase (PPO) about 100% when compared to the control. they have always been found to be less effective than useful in inhibiting browning reactions in stored apples. Kasetsart University. acids to produce complex brown polymers. They found that ficin and oxidase (1). or catechol on enzymatic browning inhibition.4252 J. which acted as strong. which show competitive properties ‡ Oregon State University. Agric. hexylresorcinol beyond 0.1) is Labuza et al. respectively (9). acidulants. Sulfites act as PPO free extract prepared from the fig latex can inhibit enzymatic inhibitors by serving as reducing agents and also react with intermediates to prevent pigment formation.18. there were some additional 10. toward endive PPO. however. This consumer awareness commercial papain preparations (from papaya latex) contain has stimulated the search for natural and safe antibrowning some substances that act as “quinone-trapping” compounds. Martyniuk reported that polyphenol oxidase (PPO) in the presence of oxygen. † Kasetsart University. pineapple juice showed browning inhibition to a similar extent as 8 mM ascorbic acid but less than 4 mM sodium metabisulfite. consumers are concerned about the possible able flavor effects.03% might result in undesir- Nowadays. potatoes and apples. Oregon 97331 The effectiveness of pineapple juice in enzymatic browning inhibition was evaluated on the cut surface of banana slices. sulfite is used extensively because of its apples during storage at 4 °C. (10) showed that 4-hexylresorcinol (4-HR) was however. Gonzalez et al.

l-Ascorbic acid. pH 7.) Merr.’s farm. and various inhibiting solutions (H2O. and b* (yellow Chemical Analysis. The Inhibition Study. to provide a PPO activity of 20-30 units/ a modified method of Ngalani et al. and preventing enzymatic browning in banana puree. pineapple % inhibition ) [(activity of control - juice. VA) to ultrafiltration following the method of Ota et al. on the effectiveness The neutral phenolics and acidic phenolics from each C18 cartridge were of pineapple juice as an inhibitor of enzymatic browning in then eluted with 100 mL of methanol and were defined as the NP bananas. Banana pulp (25 g) was defined as F2.Browning Inhibition of Banana by Pineapple Juice J. and F3 fractions) at 25 °C. The supernatant was filtered through a Ewart (22).0. and the residue was dissolved in 20 the fraction that was most effective for banana PPO inhibition mL of water to test for enzymatic inhibition. Pineapples were peeled and crushed were eluted with water. lyophilized. however. Bananas with similar sizes and no external defects enzyme. (14) who found that with a resistivity of 18 MΩ cm as ASTM standard specification for pineapple juice and ion-exchanged pineapple juice were as reagent water type I (Millipore Corp. pH 7. The percent inhibition was calculated as follows: sliced into pieces approximately 3 mm thick and 7. The first effluent was rotary evaporated at 35 °C to for 30 min. and NP increase in absorbance at 475 nm. DuPont. (18) using dopamine as the substrate. containing 1% (w/v) polyvinylpyrrolidone and 0. pH 2. following the method of Jayaraman fractions and F1. permeate. Three milliliters of the DE fraction was adjusted variety) were obtained from Dole Inc. adjusted with 0. The objectives of this study were to fractionate fraction and the AP fraction. Then.] is an important fruit ultrafiltration cell (Amicon Inc. This fraction was by a modified method of Galeazzi et al. Sigma.45 µm membrane filter. Louis. Uppsala. The PPO activity was determineded by measuring the buffer. Inc. This fraction was pH 6. Organic acids and anionic substances were retained.0 with 6 N HCl and applied to the column. and 100 mM 4-HR). Fractionation by Ion Exchange Chromatography. The lyophilized pineapple juice was dissolved in distilled in water. C18 cartridge was defined as the directly eluted fraction (DE fraction).5. conduct compositional analyses. 55. They were measured the substrate. adjusted with 1 N HCl and was filled in Thailand were chosen at ripening stage 5 (yellow peel with green in a polyprep column (Bio-Rad Laboratories..1 N NaOH. The cationic harvested at maturation stage 4 (shell color is on-half to three-fourths substances were retained. DE. Deerfield. of 6 mL).5 mL of 5 mM dopamine (Sigma Chemical S-200 HR column were pooled. water was mixed with the same substrate solution before adding the tion of Banana. and 0. They were peeled and cross-sectionally cut with a enzyme responsible for a change of 1 absorbance unit at 475 nm/min stainless knife at the head and the end of fruits and then longitudinally at 25 °C. pH 2. (Sorvall RC 5 C Refrigerated Centrifuge. The crude banana PPO was extracted elution was continued with 20 mL of water. CDU) was just after immersion (0 h) and after storage at 15 °C for 6.0 with 0. Banana slices were treated by immersion in distilled water (control). Heto Lab Equipment. 0.0 (18). The L* (lightness). Newtown. and filled in the polyprep column for a total volume water to obtain 12 °Brix as fresh pineapple juice before use. organic acids and other anionic substances were homogenized with 50 mL of a cold 0.. cartridge was preconditioned for neutral phenolic adsorption by The use of pineapple juice to inhibit enzymatic browning was sequentially passing 50 mL of methanol and 50 mL of Milli Q water investigated by Lozano-de-Gonzalez et al. Cation exchange resin (SP Sephadex C 25. 100 mL of juice permeate was using various size and charge separation methods.0. 2007 4253 efficient PPO-inactivating agents whose structure and inactiva. pH 7. Pineapples (Smooth Cayenne total volume of 6 mL. The clear juice was lyophilized in powder form by a freeze 3 mL and adjusted to pH 7. pH 4. The juice was then centrifuged at 19000g was defined as F1. The elution was continued with 20 mL of gold) as Dole’s fresh pineapple color standard guide. MO) substrate solution in 20 mM sodium phosphate banana PPO.5. This fraction to juice with a hydraulic press. F2.1 N HCl and passed through the second acidic C18 cartridge to adsorb pineapple juice was the major inhibitor of enzymatic browning acidic phenolic compounds. for acidic phenolic effective as sulfite in enzymatic browning inhibition of fresh adsorption. and organic acid compositions were .0 with 0.1 N NaOH and passed through the results were achieved with a cation-exchanged fraction.5 mL of each tested fraction (AP. eluted with 20 mL of 1% formic acid in methanol. the other cartridge was preconditioned by passing 50 mL and dried apple rings. Agric. The Banana PPO Preparation. Sweden) as phosphate buffer. and retentate of pineapple juice after UltraScan XE (Hunter Associates Laboratory. There are no published reports. pH 7. 12. (16). Vol.1 M sodium phosphate buffer. and identify evaporated at 35 °C to dryness. The permeate was then crop in many tropical and subtropical countries that is consumed further fractionated by solid-phase extraction using a C18 cartridge (total volume of 20 mL. One unit of enzyme activity was defined as the amount of were selected.. Food Chem. and then. defined as µg of tyrosine produced per min at 37 °C. 10 mg of lyophilized banana PPO (0.5 mL of Effectiveness of Pineapple Juice in Enzymatic Browning Inhibi. Denmark) for use throughout anion exchange resin (DEAE Sephadex A 25. 4 mM sodium activity of treatment)/activity of control] × 100 (1) metabisulfite. The change in et al.6. 10.0.01 N HCI instead of water.0. Treated banana slices were drained and packed in a plastic bag for storage at 15 °C.5% Triton defined as F3. pH 7.06 mg protein/mg) from ammonium sulfate saturation followed by conventional column chro. Sulfhydryl contents were determined by the method of at 10000g (4 °C) for 30 min. 48. Wen preconditioned neutral C18 cartridge to adsorb the neutral phenolic and Wrolstad (15) reported that a nonvolatile organic acid in compounds. a* (red to green color dimension). Richmond. Total and 72 h. Sephacryl-S 200 HR was dissolved in 2 mL of 20 mM sodium matography on Sephacryl S-200 HR (Pharmacia. Pineapple juice is a fractions following the method of Jaworski and Lee (19). Then. CA) for a tip) according to the peel color index. The pineapple juice was centrifuged Singleton (21). water. Bananas [Musa (AAA Group) Gros Michel] grown preconditioned in water. (20) using casein as evaluate the browning inhibition of each treatment. this fraction was further fractionated by ion exchange chromatography MATERIALS AND METHODS technique. 10000 Da molecular mass cutoff ultrafiltration membrane with an Pineapple [Ananas comosus (L. Thailand. MA). phenols were analyzed using the modified method of Slinkard and Pineapple Juice Fractionation. Each of the three fractions was rotary evaporated at 35 X-100. preconditioned the study. They were to pH 2. cationic substances Pineapple Juice Preparation. 8 mM ascorbic acid.. Twenty microliter aliquots of the enzyme were added least 30% of the PPO activity of the most active fraction from Sephacryl to the mixture containing 2. MA). No. Bedford. pH 7. and the residue was dissolved in 3 mL of water. Reston. The effluent portion was then adjusted to pH 2. absorbance at 475 nm was recorded for 1 min..0. St.0. IL) into three fresh or in various processed forms (12). The three fractions were then rotary pineapple juice. adjusted with 1 N HCl. Alltech Associates Inc. 24.5 cm long. Because the DE fractions from solid-phase extraction showed the highest inhibition. Beverly. and used as a source of Co. The fractions containing at mL of enzyme.8. All determinations were performed in triplicate. CT). One unit of enzyme (casein digestion unit. first. 0.. and the filtrate was ultrafiltered through a tion mechanism remain to be elucidated (11). The bromelain activity was determined in the to blue color dimension) values of the cut surfaces were measured by pineapple juice.5 with 0. (17). After pineapple juice was fractionated of 0. the popular product due to its very pleasant aroma and flavor (13). sugar. The homogenate was centrifuged for 30 min (4 °C) at 29000g °C to dryness. The effluent from the preconditioned acidic in apple products.1 N NaOH and then applied to the dryer (Heto FD 2. the best adjusted to pH 7. Sigma) was Plant Materials. As a control.0. pH 7. To test the relative inhibition caused by each crude enzyme was then partially purified by fractionation with 80% fraction.

Alltech Associates Inc. 10.6 65. 6. MA). 12. 4-HR. Isocratic elution with 82% (v/v) tissue. The puree was then filled in plastic melain activity (11. The DE fraction had significantly more inhibition than Because controlled banana slices and banana slices treated with other fractions and resulted in 100% PPO inhibition as compared distilled water. was used at a flow rate of 0. 1. 4 mM sodium metabisulfite. and detection was measured at 214 nm.4254 J. isocratic elution with 0.1 M potassium dihydrogen phosphate. For L-ascorbic acid and organic acids analyses. had the lowest of Banana Puree. not surprisingly.0 99.5. The DE fraction was further 3 days. Each enzymatic browning inhibition on the cut surface of banana slice fraction was concentrated five times as compared to the juice was evaluated and compared to various inhibitors (Figure 1). With respect to banana slices treated with 100 mM fractionated by ion exchange chromatography. AP. Therefore. The ultrafiltration was performed for only a short time (3-4 Milford. only the to the control (Table 2).6 0.0. The AP fraction indicated that pineapple juice was as effective as 8 mM ascorbic had a higher sulfhydryl content than the NP fraction but seemed acid in inhibiting browning of banana slices stored at 15 °C for to have no inhibition effect on PPO.78 CDU/g of pineapple juice powder) containers and the L*.5 20. IL) statistical analysis al. This indicated that the major polyphenolics in treated with 8 mM ascorbic acid as well as those treated with pineapple juice were neutral phenolic compounds that could pineapple juice showed similar decreases in L* values. total phenols SH content determined by high-performance liquid chromatography (HPLC) using pH °Brix (ppm) (µM) % inhibition the methods of Wen and Wrolstad (23).) were connected in series NP 7. pH 2. the HPLC system consisted of a 4100 apple slices with 4-HR at concentrations beyond 0. However. Labuza et Statistical Analysis. Tag Chemistry Package (Waters. AP 2. Banana slices inhibition. permeate.2 349 22.4 c to a Dynamax model SD-300 pump and a HP 1040 diode array detector. Tag reagents and a h). 50%) to the same extent as pineapple the following test solutions: water with the pH adjusted to 2. Effect of Fractions Separated by Solid-Phase C18 Cartridge Figure 1. Free amino acids were determined by derivatization tration was used to remove compounds greater than 10000 Da. pH 4. This inhibit enzymatic browning to some extent. and 24 h of storage time at 5 °C. The DE fraction contained higher L* values of banana slices treated with distilled water. Glucose. and reported as g total Effectiveness of Fractions from Pineapple Juice. pH 4. For organic acids.d. L* value of banana slices treated with pineapple juice and on Banana PPO Inhibitiona various inhibitors during storage at 15 °C for 3 days. browning of amounts of total soluble solids (5. it can be concluded that the protease activity or bromelain in The purees were left to reach room temperature before measurement. (7) also reported that bromelain did not inhibit mushroom system was used for analysis of the data. The NP fraction. which consisted of Waters Acc Q.2 mL/mm. the Effectiveness of DE Fraction on Enzymatic Browning Inhibition permeate of pineapple juice. and the level of . 55. with 0. and b* values of the puree were measured showed the lowest value in %PPO inhibition (27. columns ODS-2 and ODS-1 (250 mm × 4.4 11.9 12.0.03% (w/v) LDO pump and a RI 4 LDC detector. which contained low for all treatments decreased with time.6 mm i. The statistical significance PPO activity in an aqueous model system. Significant effective in the browning inhibition of refrigerated apple. Effect of Pineapple Juice and Ultrafiltration-Treated Juice on Banana PPO Inhibitiona Bromelain activity pH °Brix (CDU/g PJ powder) % inhibition pineapple juice 3. Vol. they exhibited a turbid white appearance and had an Malic and citric acid contents were determined and reported as g total unusual odor.4 °Brix).4. amounts of total soluble solids and sulfhydryl content than the are shown. molecular mass of bromelain is more than 30000 Da (25). Phenolic profiles were determined using the method higher than that for the pineapple juice and permeate. Banana puree was prepared by blending 30 g of level of bromelain activity as shown in Table 1. banana pulp for 30 s with 20 mL of water (control) and with 20 mL of it still inhibited PPO (c.. it was difference was assessed by one-way analysis of variance.7 b a Means within the same column with different letters are significantly different (p e 0. a*.7 a DE 2.1% dithiothreitol at a flow rate of 0.7%). influenced apple flavor due to the residual content in the apple NH2 (250 mm × 4.05).05) among treatments were detected using Duncan’s McEvily (8) confirmed that ficin from fig latex did not affect multiple range test. Effectiveness of Pineapple Juice in Enzymatic Browning The juice permeate was further fractionated into three Inhibition of Banana. acidic phenolic fraction.7 mL/mm.2 40. Food Chem. had the highest banana slices treated with 100 mM 4-HR and 4 mM sodium concentration of total phenols and gave about 65% PPO metabisulfite were lower than other treatments.0 1.3 a permeate 3.0 (as the juice. method (24) using the Waters Acc Q. fructose. neutral phenolic fraction. Agric.1 5. and NP. pineapple juice was not involved in PPO inhibition.04 52. and DE fraction.9 b 5 µm particle size. summed.a. For sugar analysis. Ultrafil- sugars/100 mL. the mobile phase was 0. total soluble solids of the retentate was a little C18 column. For L-ascorbic acid. acetonitrile in water was used at a flow rate of 0. min.7 mL/ (p e 0. As the described by Wen and Wrolstad (23). The column used was Zorbax.5 122 7.0 12. therefore.9 11. Nevertheless.6 mm id).05). No. Monsalve et al. however. pH 2. and sucrose were determined. The SPSS (Chicago. the activity of mushroom PPO but the protein-free composition inhibited the enzymatic and nonenzymatic browning of foods RESULTS AND DISCUSSION and beverages susceptible to browning.78 27. browned to a similar extent.1 a retentate 4. 8 mM ascorbic acid. differences (P e 0.15 49. 2007 Chaisakdanugull et al. 3..5% potassium dihydrogen a Means within the same column with different letters are significantly different phosphate. the retentate that contained the highest bro- same pH of DE fraction). Moreover.4 3. at 0 (initial condition). The results indicated that L* values of banana slices NP and AP fractions. and detection was measured at 245 nm. Table 2. Table 1.4 923 3. The effectiveness of pineapple juice in fractions by solid-phase extraction using C18 cartridges. (10) also found that treatment of acids/100 mL.

0 b F3 2. 20% sucrose.2 g % Su (F2) 1. and organic acids.5 g % Org acid + 14. c Total sugar is the sum of sugars (glucose.1 (g/mL) Tyr (F2) 27. 4. known as a phenolase Cu-chelating agent. 2.3 0 87. Table 4. The F2 fraction contained more activity. Effect of Fractions Separated by Ion Exchange Column on Banana PPO Inhibitiona totalb acid total amino Cys Tyr ascorbic acid totalc sugars total phenols SH content pH (g/100 mL) acid (g/ mL) (g/mL) (g/mL) (mg/100 mL) (g/100 mL) (ppm) (µM) % inhibition F1 8. and fructose) determined by HPLC.8 b a Means within the same column with different letters are significantly different (p e 0. Of the amino acids. the and expected to be inhibitors of PPO. cysteine.8% inhibition. Agric. consisted of negatively charged compounds. and the inhibition of The F2 fraction.8 g % Org acid 54.1 (g/mL) Tyr (F2) 0. ways: by reacting with the o-quinones and by chelating the essential copper at the active site of PPO. tyrosine.1 0 3. Effect of Various Compounds Found in the Separated Fractions by Ion Exchange Chromatography on Banana PPO Inhibitiona solution % inhibition 27.1% malic acid and 0. sucrose. Cys.2 104 11.6 c 27. 2007 4255 Table 3.4 0 0 1 0 3. fraction from pineapple juice. p-coumaric acid. the exchange resin.0 (g/mL) Cys (F2) 6. However. Citric acid is also browning inhibition. It is generally accepted that thiol compounds inhibition of F1 indicated that positively charged compounds block the enzymatic browning reaction by forming colorless did not effectively contribute to pineapple juice enzymatic addition compounds with o-quinones (27).2 21.2 g % Su + 0. Su. The results show that sugars and tyrosine (major peak in no ascorbic acid and only a trace amount of sulfhydryl phenolic profile of F2) were not effective inhibitors of PPO. The lowest level of PPO tyrosinase (26). Food Chem. This suggests a minor major components of pineapple juice that are responsible for role for cysteine in PPO inhibition of F2. tryrosine. organic acid (0. and F2.7 36. neutral phenolic fraction.41 1.5 g % Org acid + 16. Ascorbic acid is known to inhibit PPO. F2.2 g % Su + 0.03 total 0 0.5 1550.2 2. Therefore. 3. serotonin. cysteine.6 41 51.1 d 27.7% glucose. a mixture of sugars. F2. sugar (6. and other sulfhydryl compounds.1 0 0 12 3. phenolic compounds in F2 were composed of tyrosine and To further test the effect of the possible inhibitors on PPO serotonin as shown in Figure 2.5 g % Org acid (F2) 14. organic acids. No. Chromatograms of phenolic compounds from pineapple juice fractionation tentatively identified by HPLC: 1.2 g % Su+ 0. in Table 5. 55. inhibitory effect of F2 may be due to the combined effect of Organic acids were present in low amounts in this fraction. The compounds.7 a F2 5. The organic acids.5 a 2. compounds were found in F3.0 (g/mL) Cys (F2) 27. which were detected in F2 and F3 and total amino acids than the other fractions. Combinations of sugars enzymatic browning inhibition are organic acids. Organic Acid Contents of Fractions Obtained from Ion Exchange Column concentration (g/100 mL) organic acids F1 F2 F3 malic acid 0 0.80 citric acid 0 0.13 1.4% citric acid).6% fructose). model system experiments were conducted as shown sugars.2 0.0 (g/mL) Cys (F2) 51.Browning Inhibition of Banana by Pineapple Juice J.2 g % Su + 0.05).8 193. which was directly eluted from anion PPO was attributed to the chelating action (28).1 0 0. F1. and NP. and F3. Vol.5 g % Org acid + 12. were prepared at the same level of PPO inhibition of F2 was lower than F3. sulfhydryl compounds.6 a 41.83 Table 5.05).0 c 41. Tyr. DMHF (D-methylhydroxylfuranone).1 0.4 27. fractions obtained from the pineapple juice DE fraction. Figure 2. fraction from Amino acids are known to affect PPO activity in at least two pineapple juice. which concentration of F2 except that the organic acid solution (malic consisted mainly of organic acids (malic and citric acids) as acid and citric acid) was prepared at the same concentration of shown in Table 4. and 0. Org acid. The fraction F1 consisted largely of browning in avocado and banana tissue as well as of mushroom positively charged compounds. and .0 c 51.0 c a Means within the same column with different letters are significantly different (p e 0. 10.4 c 51.54 2. total phenolics. ascorbic acid. PPO inhibition achieved by the fractions and their compositions L-cysteine was shown to be the most effective inhibitor of are shown in Table 3. These results indicate that the Cysteine alone resulted in 6.1 (g/mL) Tyr + 41. b Total acid is the sum of organic acids (malic acid and citric acid) determined by HPLC. but F3..

. S. which is less than the 21. T. J. Corvallis. Technol. and polyphenol oxidase by proteolytic enzymes “Killer Enzymes”.1%) was shown by the organic acid fraction and Their Effect on Health. DE fraction (Table 6). J.... 1982. Nutr. guidance. The fraction that (18) Jayaraman. Oregon State University. 1998. Paris. pH 2. Richard-Forget. T. and comparison with manual methods. 1977. R.: New York. neither like nor dislike.. 1999a. fractionation of the DE fraction by ion exchange chromatog- (17) Ngalani. Further polyphenoloxidase (PPO) from a Dwarf variety of banana (Musa CaVendishii. Lee. Sgarbieri. 95-100. 1990. Inhibition of enzymatic (23) Wen. Enzymatic browning inhibited in fresh and dried In conclusion. 1981... 47.9 b 63. Isolation and characterization of a ‘quinone- inhibitory effect. 49-55. E. Therefore. Technol. 1987.. 1993. P. 6. (3) Sapers. Lillemo. 55. J. Signoret. L. J. Meeting inhibitory effect was not caused by the bromelain activity. 1993. R. Federation of Fruit Juice Producers. E. (20) Ota. Ho. L. the color acceptability score of the samples Food Eng. Abstr. e 0. F. Partial purification and raphy along with model system experiments indicated that the properties of plantain polyphenol oxidase. 677-684. J. Agric.3 a 4.708. Crit. 66.981. dislike extremely. K. C. D. G. H. M. 1994. Vitic. Cerny. V. P.) Merr. (8) McEvily. While banana puree treated with sulfite had a higher (13) Rattanathanalerk.1 a 64.. Browning of food: Control by sulfites. G. Fayad. inhibition of banana slices stored at 15 °C for 3 days. inhibition of pineapple juice.. Cerny.. Durst.. 35. Y.. like moderately. J. 1991. 259-265. W. Singleton. Effect L* value (lighter color) after 3 h at 5 °C than puree treated of thermal processing on the quality loss of pineapple juice. 1993. Wrolstad.4 b 70. Wen and Wrolsad (15) reported (10) Monsalve-Gonzalez. P. M. Lee. Biochemistry Science and Technology.. (5) Oszmianski...]. R.. Steven. Wrolstad. concentrate.. Patent 4. dislike moderately.. Polyphe- Treated with DE Fraction and Various Antibrowning Solutions after noloxidase by a Carica papaya latex preparation.. 2. Srichumpoung. P. T. Food Chem. Nauk. Food Chem. 1892-1895. 8.7 d istry 1989. Food Sci. similar results in that most of the browning inhibition activity P. N. S. N. LITERATURE CITED (22) Ewart. Gross. A. Saunier. M. A. S. R. 4-hexyl- whose structure was not completely characterized..0% inhibition of F2. 75-84. S. (6) Martyniuk. 47. and 9... M. Inhibition of Banana Puree. No. (7) Labuza. J. 2007 Chaisakdanugull et al. C. Food Technol. J. M. V. Huang. ReV. Eds. for providing pineapples for the research. Taoukis. Color acceptable scores: 1. 46. aghavan. Abstract No.. Inhibition of polyphenol present in F2 that could inhibit PPO activity. Vijavar- contained cysteine and other sulfhydryl compounds also con. W.. with the DE fraction. like slightly. abisulfite. Table 6. 36. J. R. showed that the water at pH 2. A. Int. R. factors. 182-185. 3.. 1961. The highest level oxidase by phenolic compounds. J. Food Sci. Food Storage at 5 °C for 3 ha Sci. C.0 acid sulfite fraction (4) Janovitz-Klapp. 113-118. Preparation and chemical We thank Bob Durst and the staff at the Department of Food properties of purified stem and fruit brimelains. 399- was as effective as 8 mM ascorbic acid in enzymatic browning 404. lyengar. 58. 1992. Y.. (12) Avallone. Cereal Chem. gave the highest (16) Galeazzi. The preliminary experiment (11) Richard-Forget. P. S. S. Fractionation and HPLC determination ACKNOWLEDGMENT of grape phenolics. Food Sci. Guiraud. Lee. W. J. C. 28. J. 58.. Canalieri. partial purification and some properties. (1) McEvily. 435. Brillouet. Nicolas. Research Report... Curr. Wrolstad. The 1999b. which contained polar compounds. R. like extremely. A. G. Food Chem... Barbosa-Canovas. K. J..9 c from apple. John Wiley and Sons Inc. Food Sci. Some inhibitors of polyphenoloxidase in honey. Y. K. Oregon State University. purification and physicochemical characterization of acid in enzymatic browning inhibition of banana puree. Technol. Pszczelinicze Zesz. Food Chem. 32 (3). Automation Thailand. U. J. Total phenol analysis.. McEvily. 1999 IFT Annu. D.. A. 1993. 19. Symposium on The 20th International Symposium International tyrosine inhibited PPO to the same extent. M. M. Enol. Constantinides. 10. Food Sci.4256 J. P. Special thanks go to Dole Inc. C. J. E. H. C. Phenolic composition of pineapple juice browning in foods and beverages. L* Values and Color Acceptable Score of Banana Puree (2) Rigal. Barrett.0 did not exert a browning Varoquaux. J. S. A. A. treated with sulfite was lower than for those treated with the (14) Lozano-de-Gonzalez.. 2003. Agric. A. and friendship. J. S.. 1988. Agric.. 67-73. M.. Vol. in Table 6. Studies on disulfide bonds in glutenin. F. 3. cysteine.. Chiewchan. 2005. Gouzet. J. J. J. and a mixture of sugars. our results have shown that pineapple juice apple rings by pineapples juice. M. 11A-15A. p 8.3 b 6. R.. Inhibition of cysteine. Iyengar. 33. therefore. 38. 285-296.. Iyengar. 797- Effectiveness of DE Fraction on Enzymatic Browning 800.. A.. 257-259.. The (15) Wen. D. A. pp 235- the important role of organic acids in enzymatic browning 340. P. dislike slightly. A. dislike very much. 50-155. the low pH of the DE fraction is trapping’ substance from a crude Carica papaya protein prepara- not believed to be responsible for browning inhibition.05). 341-347. . M. 253-273. T. Teisson. DE fraction. V. (21) Slinkard. Polyphenoloxidase L* values 62. Phytochem- color acceptable score 3. played an important role in PPO inhibition.. Inhibition of polyphenoloxidase a Means within the same row with different letters are significantly different (p activity and browning by honey. antioxidants H2O 8 mM ascorbic 4 mM DE and other means. 65. In Phenolic Compounds in Food of inhibition (54. Richard. Y. J. having the same concentration as F3 (Table 5). M. like very much. Control of browning during in pineapple juice was caused by a nonvolatile organic acid storage of apple slices preserved by combined method. Microbiol. Am. 28. for their help. A. (19) Jaworski. This confirms T. L. 7. L). 4. Stein. 38. J. resorcinol as an antibrowning agent. Int. main organic acids (malic and citric acids) in pineapple juice 48. Pap. Enzymatic browning in some banana varieties tributed to the effectiveness of pineapple juice in enzymatic as related to polyphenol oxidase activity and other endogenous browning inhibition. 181-186. acid but lower than those treated with 4 mM sodium met. L* values of banana puree treated with the DE Enzymatic browning and biochemical alterations in black spots fraction were higher than for those treated with 8 mM ascorbic of pineapple [Ananas comosus (L. organic acids.2 c 7. 2001. N. Food Sci. 5. 2903-2907. percent PPO inhibition and was more effective than ascorbic Isolation. about 14% inhibition. Moore.S. Varoquaux.. As shown tion. F. Ramanuja. Dhakne. M. W. compounds in addition to organic acids and cysteine may be (9) McEvily. 1990. S. 1992.

M. J. Food Chem. chem. J.. Y. A. Yasuda. 1993. We express sincere gratitude (26) Kahn. Purification tizing reagent. Revised manuscript received 52. Cysteine as and Research in Agriculture (SEARCA) for the financial support. an inhibitor of enzymatic browning II. Sci. Agric. S. 111-115. Yasui. Synthesis of a fluorescent deriva. of polyphenol oxidase and the browning control of litchi fruit ate. 211. J. Kinetic studies. (25) Murachi.. 40. 1992. 2007. Michau. 79.. F. 55. Food Chem. 3. Anal. 48. 2007 4257 (24) Cohen. Y.. Agric. March 14. Food Agric. avocado and (KURDI). 50. Effect of protein. Zauberman. isters of Education Organization Regional Center for Graduate Study (27) Richard-Forget. Biochemistry 1994. Kasetsart University. P. Fu. Goupy. Bio. M. T. Nicolas. Vol. 954. D. M. Accepted March 16. 6-aminoquinolyl-N-hydroxysuccinimidyl carbam.. 279-287. J.. G. No.. (28) Jiang. 2108-2112. and Southeast Asian Min- banana. 1985. JF0705724 . J. Fuchs. 950- acid via high-performance liquid chromatography. protein hydrolysates and amino acids to the Kasetsart University Research and Development Institute on o-dihydroxyphenolase activity of mushroom. 10. 1999.Browning Inhibition of Banana by Pineapple Juice J.. V. Purification and physical characterization of stem bromelain. Thailand. M.. Y. 2007. Food Sci. 2007. and its application for the analysis of the hydrolysate amino by glutathione and citric acid. P. R. Received for review February 27. J.