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EFFECTS OF COMMERCIAL PECTINASES APPLICATION ON SELECTED PROPERTIES OF RED PITAYA JUICE

A.R. NUR ALIAA, M.K. SITI MAZLINA1 and F.S. TAIP Department of Process and Food Engineering Faculty of Engineering Universiti Putra Malaysia 43400 UPM Serdang Selangor, Malaysia
Accepted for Publication February 19, 2009

ABSTRACT Pitaya or dragon fruits are becoming more popular in Malaysia due to their nutritional benets. These exotic fruits can be utilized to produce many functional products such as fruit juice. This work examined the effects of commercial enzymes, i.e., Pectinex Ultra SP-L and Pectinex CLEAR on the chemical composition, vitamin C and total polyphenols contents of red pitaya (Hylocereus polyrhizus) juice, which is the most popular variety of pitaya. We have also observed the differences of these properties in fresh fruit pulp and juice produced without enzymatic treatment. The pitaya juice produced after enzymation and pasteurization has not shown marked changes in the most of its major chemical parameters such as moisture, ash, fat, carbohydrate and calorie. However, enzymation using Pectinex CLEAR leads to the juice with higher yield of protein. The protein content after the treatment increased to 2.20% w/w from 0.23% w/w of the raw juice. Vitamin C content in treated samples had reduced slightly because of the heat treatment. However, phenolics amounts were slightly higher up to 7% in the treated samples which suggest that pitaya beverage is rich in antioxidant capacity than in the unprocessed fruit. This study demonstrated the effects of pectinases application on several properties of red pitaya juice with an attempt to obtain some interesting and valuable data for the fruit processing industries. The data obtained were helpful for the optimization of red pitaya juice production as a step toward promoting the potential of red pitaya juice as a functional beverage.

Corresponding author. TEL: +00-603-89466363; FAX: +00-603-86567123; EMAIL: siti@ eng.upm.edu.my

Journal of Food Process Engineering 34 (2011) 15231534. All Rights Reserved. Copyright the Authors Journal Compilation 2009 Wiley Periodicals, Inc. DOI: 10.1111/j.1745-4530.2009.00388.x

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PRACTICAL APPLICATIONS This paper aims to provide some useful data for pitaya or dragon fruit juice production. Since not much work has been done on this, the aim of this study was to determine the effects of commercial enzymatic treatment on clarication of red pitaya juice. Pectinase enzymes have been used in this study to help improve the quality of the juice physically and at the same time preserve some of the important properties of the juice such as vitamin C and total polyphenols contents. The process developed is considered suitable and very helpful for the processing of pitaya or dragon fruit processing.

INTRODUCTION Pitaya or dragon fruit is known to be highly nutritious. This exotic fruit is being converted into ready-to-drink beverages, jam and wine. Currently this fruit utilization is further expanded to produce dragon fruit enzyme and dragon fruit concentrated juice. These products are now available in the market in Malaysia. This expansion has promoted dragon fruit as a great source of functional beverages. However, less attention is being given to the appearance and texture of the drinking products. The texture of the so-called concentrated juice causes difculties in mixing the concentrate with water to be drunk. This situation may not attract the consumers to buy this product again. Hence, action should be taken to overcome this problem. Juice with low viscosity, high clarity and yet high in nutrition is more desirable for consumers. Enzymes can play a key role in improving juice clarity, stability as well as reducing the viscosity. Enzymes are being used to clarify and stabilize the juice and degrade soluble pectins and starches that cause haze. Enzymatic treatment using pectinase is an effective way to reduce the pectin in the fruit juices because pectinase has the ability to hydrolyze pectin and cause pectin-protein complexes to occulate (Rai et al. 2004; Lee et al. 2006; Liew Abdullah et al. 2007; Sin et al. 2006). Pectinases are obtained from plants and fungi. However, the major commercial source of pectinases is Aspergillus sp. Pectinases are used industrially to weaken plant cell walls and enhance extraction of fruit juice. They help to improve juice recovery, decrease viscosity and prevent gelation in the extracts. The degradation of plant cell walls by exogenous enzymatic treatment results in easier release of the components contained in cells (Janser 1997). Fruit and vegetable juice production is nowadays unthinkable without the use of enzymes (Baumann 1981). A complete enzymatic breakdown of pectin is the key to producing clear and stable fruit juice. Generally, fruit juice extraction involves maceration of the pulp followed by centrifugation and

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ltration, to separate the juice from the solids. The aim of mash treatment is to increase juice yield and to facilitate the extraction of the soluble substances from the cells (Baumann 1981). Treatment of fruit pulp with appropriate enzyme preparations is a general practice in juice processing (Ramadan and Moersel 2007). Filtration is employed to remove insoluble plant material from the enzyme-treated juice. For a few years back, researches have been done to study on the color pigments of red pitaya (Stintzing et al. 2002; Mobhammer et al. 2005; Herbach et al. 2006; Esquivel et al. 2007). Research on the cultivation of pitaya has also been done by Zee et al. (2004). The general chemical composition of red pitaya has been reported (Ilovepitaya.com 2007). In this work, the chemical composition, vitamin C and total polyphenols contents of fresh and treated red pitaya juice are reported for the rst time. The data obtained are helpful for the optimization of pitaya juice production as a step toward promoting the potential of pitaya juice as a functional beverage. MATERIALS AND METHODS Materials Ripe red-eshed pitaya (Hylocereus polyrhizus) were obtained from a local farm in Sepang (Selangor, Malaysia). Two commercial pectinase enzymes preparations were applied: Pectinex Ultra SP-L (from Aspergillus niger) and Pectinex CLEAR (from Aspergillus aculeatus and Aspergillus niger) both obtained from Novo Nordisk Ferment Ltd., Dittigen, Switzerland were used for enzymatic treatment of red pitaya juice and stored at 4C. Pectinex Ultra SP-L and Pectinex CLEAR are commercial enzymes used to produce clear and stable fruit juices. The commercial enzyme contains mainly polygalacturonase, pectinesterase and pectin transeliminase. The activity of Pectinex Ultra SP-L and Pectinex CLEAR enzymes are 26,000 PG/mL (polygalacturonase activity per mL). It appears that such enzymatic preparations rupture the juice retaining particles of the pulp, such effect being apparently due in part to the action of polygalacturonase in splitting the peptic chain and the action of pectinesterase in hydrolyzing the methyl ester groups of the pectin molecule (Wieland 1972; Landbo et al. 2007). The optimum enzyme reaction conditions are at pH 3.56.0 and temperature range below 50C (Kashyap et al. 2001). These are ideal conditions for fruit processing. Methods Production of Claried Red Pitaya Juice at Laboratory Scale. Pitayas were peeled and extracted using a juice extractor (Philips Juicer Model:

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TABLE 1. OPTIMUM PROCESSING CONDITION FOR ENZYMATIC CLARIFICATION OF RED PITAYA PULP Types of enzymes Enzyme concentration (%) 0.10 0.09 Incubation temperature (C) 40 46 Incubation time (min) 45 82

Pectinex Ultra SP-L Pectinex CLEAR

HR 1858155; Royal Philips Electronics, Eindhoven, The Netherlands) to remove the seeds and coarse cloud particles. Two commercial enzyme preparations were added separately to the pitaya pulp. The enzyme concentration, time and temperature of enzymatic treatment for Pectinex CLEAR enzyme have been obtained from trials done by Siti Mazlina et al. (2007). For Pectinex Ultra SP-L, the processing condition data for enzymatic clarication of red pitaya juice was also obtained from trials (data not published). For each sample, Pectinex Ultra SP-L and Pectinex CLEAR were added separately to the pulp and incubated (Wise Cube Shaking Incubator Model: WIS-S10; DAIHAN Scientic, Co., Ltd., Seoul, Korea) with continuous shaking (250 rpm). Table 1 shows the optimum conditions for enzymatic clarication of red pitaya pulp using both enzymes, respectively. The treated pulp was pasteurized at 90C for 5 min in a hot waterbath (Laboratory Waterbath Model: LMWB-6PC; PLT Scientic Sdn. Bhd., Selangor, Malaysia) and then cooled at 2C using circulating waterbath (Model: 71; PolyScience, IL). These steps were performed on the samples to inactivate the enzyme action before ltration. The treated red pitaya juice was centrifuged (Refrigerated Centrifuge 5800; Kubota Corporation, Fujioka, Japan) at 3,000 g for 10 min, and the supernatant was collected. The juice was ltered through a lter paper (Whatman No.1, Whatman International Ltd., Kent, England), which has the pore size of 11 mm using oil-less vacuum pump (Model: Rocker 300; Rocker Scientic Co., Ltd., Taipei County, Taiwan). The separation of the pulp from the juice by ltering the liquid pulp on a rotary vacuum lter avoids the costly pressing step and the juice obtained is clear (Baumann 1981). Figure 1 illustrates the step for the production of claried red pitaya juices at laboratory scales. A control sample was produced without enzymatic treatment step but was pasteurized at 90C for 5 min followed by cooling at 2C. Determination of Chemical Composition, Vitamin C and Total Polyphenols Contents of Red Pitaya Juice. Protein, fat, carbohydrate, moisture, ash and calorie were determined according to AOAC (1995). Ascorbic

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Peeling

Pulp Extraction

Fruit Pulp

Maceration with enzyme (0.01 0.1%) for (20 100 min) at 30 50C Enzyme deactivation at 90oC for 5 minutes and cooling at 2oC

Centrifugation

Precipitate (Solid)

Supernatant (Juice)

Filtration (Oil-less vacuum pump )

Clear Fruit Juice

FIG. 1. STEPS FOR EXTRACTION AND SUBSEQUENT CLARIFICATION BY ENZYMATIC TREATMENT OF RED PITAYA JUICE

acid was determined volumetrically with 2-6 dichlorophenol-indophenol reagent (AOAC 1995). Total polyphenols was measured by modied FolinCiocalteu method using gallic acid as a standard. Five grams of juice was extracted with 20 mL mixture of methanol-water and with 30 mL mixture of acetone-water separately under stirring for 1 h. Then the supernatant were combined in volumetric ask and made up to 50 mL with distilled water and is called extract. 0.5 mL of extract is added with 0.5 mL Folin-Ciocalteu reagent and allowed to react for 3 min under continuous stirring. After that, 10 mL of sodium bicarbonate (75 g/L) was added and the mixture was diluted to 25 mL with distilled water. The solution was kept at room temperature for 2 h in the dark; the absorbance was then measured at 750 nm using UV-Vis Spectrophotometer (Model UV-260; Shimadzu Corporation, Tokyo, Japan). The mean value of total polyphenols was obtained from triplicate experiments.

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RESULTS AND DISCUSSION Treatment of fruit pulp with appropriate enzyme preparations is a general practice in juice processing (Ramadan and Moersel 2007). Enzymes are used in the juice industry to aid in the separation of juice from the fruit cells and to clarify the juice by the removal of pectin and naturally occurring starches that contribute to undesired viscosity, poor ltration and a cloudy appearance (Enzyme Technical Association 2002). Since juice extracted from ripe fruit contain substantial amount of pectins and starches, they lead to a cloudy appearance to the juice, making the juice less appealing by consumers due to the appearance and mouth-feel of the juice. The cloudiness that pectin causes is difcult to remove unless by enzymatic depectinization. Variety of enzymes available in fruit and vegetable processing contribute to some degree to the conversions, degradations and breakdown reactions desired (Wieland 1972). Depectinization affected the composition of juice in relation to the type of juice, the type of enzyme and the time considered (Versari et al. 1998). In this study, ripe red pitaya fruits were selected from a local farm in Malaysia with considerably higher quality that is reected by the fruits larger size, well-developed color and very juicy pulp. Ripe fruits lead to juice freshness and quality that is crucial to the quality of the nal product. Two pectinase enzyme application that were Pectinex Ultra SP-L and Pectinex CLEAR were examined to demonstrate the effects of pectinases on the quality of fruit juice. Effect of Enzyme Application on Chemical Composition of Pitaya Juice The inuence of different enzyme preparations on chemical parameters of red pitaya juice is presented in Table 2. By enzymatic treatment, not only the yield of juice increased, but also the macro- and micro-components contents of the product (Acar et al. 1999). The pitaya juice produced after enzymation and pasteurization has not shown marked changes in the most of its major chemical parameters such as moisture, ash, fat, carbohydrate and calorie. From the study done by Ramadan and Moersel (2007) on goldenberry juice, they found out that the effect of enzymation on these macro- and micro-constituents seems to be insignicant. It can be seen that enzymetreated and pasteurized red pitaya juice contained zero fat level. An interesting result is that the juice treated with Pectinex CLEAR enzyme preparation, resulted in markedly higher yield of protein as shown by the chart in Fig. 2. For enzyme-treated red pitaya juice, the carbohydrate content has reduced slightly. The reduction may be due to the effects of the enzymes used on the structure of red pitaya juice and hence, affected the release of microand macro-components into the juice. Carbohydrate content in other fruits was reported to be as follows: strawberry, watermelon and mango (3%); apricot,

TABLE 2. CHEMICAL COMPOSITION OF RED PITAYA JUICE Fat (% w/w) 0.00 Moisture (% w/w) Ash (% w/w) Carbohydrate (% w/w) Calorie (Kcal/100 g)

Treatment

Protein (% w/w) 0.06

Fresh fruit pulp (untreated pulp) Control sample (pasteurized only) Juice treated with Pectinex Ultra SP-L Juice treated with Pectinex CLEAR 0.20

0.23 0.00 0.00 2.20

0.10 0.00 0.00 0.00

88.40 88.43 88.97 88.17

0.00 0.06 0.06 0.06

0.50 0.50 0.50 0.60

0.00 0.00 0.00 0.00

10.77 11.07 10.53 9.03

0.06 0.06 0.06 0.15

45.00 44.00 44.00 45.00

0.00 0.00 0.00 0.00

PECTINASE EFFECT ON RED PITAYA JUICE

Values given are the mean of three replicates standard deviation. % Carbohydrate = 100 - (% Protein + % Fat + % Moisture + % Ash). Calorie (Kcal) = 4 (% Carbohydrate) + 4 (% Protein) + 9 (% Fat).

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FIG. 2. PROTEIN (% w/w) CONTENT IN RED PITAYA JUICE

TABLE 3. VITAMIN C AND TOTAL POLYPHENOLS CONTENTS OF RED PITAYA JUICE Treatment Vitamin C (mg/100 g) 6.06 3.08 4.61 3.27 0.05 0.006 0.02 0.31 Total polyphenols (mg/100 g) 1.244 1.322 1.336 1.255 0.70 0.34 0.14 0.12

Fresh fruit pulp (untreated pulp) Control sample (pasteurized only) Juice treated with Pectinex Ultra SP-L Juice treated with Pectinex CLEAR Values given are the mean of three replicates

standard deviation.

guava, lemon, orange, papaya, peach and plum (6%); apple, grape, mango and pineapple (15%); and banana and prune (20+%) (UMass Medical School 2008). Since the range of carbohydrate content in red pitaya juice was 9.03 11.07%, respectively, it can be concluded that carbohydrate in pitaya juice was higher than that of other tropical fruits such as guava and papaya, and lower than that of mango, pineapple and banana. The calories in fresh and processed pitaya juices are less than 100 kcal and can be considered as low. Thus, pitaya juice can be classied as a healthy beverage. Inuence of Enzymatic Treatment on Vitamin C and Total Polyphenols Contents of Red Pitaya Juice Table 3 shows the data obtained for vitamin C and total polyphenols content in red pitaya juice. The untreated pulp had slightly higher vitamin C content than enzyme-treated and control sample. It was observed that vitamin C content in red pitaya juices was higher than that of white pitaya juice (data not published yet). This may be due to the antioxidants available in red pitaya

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FIG. 3. TOTAL POLYPHENOLS (mg/100 g) CONTENT IN RED PITAYA JUICE

fruit. Traditionally, deep colored fruits, vegetables or foods are recognized as more healthy to human body, especially in oriental countries (Ercisli and Orhan 2008). According to the study done by Zulueta et al. (2007), they concluded that the differences between parameters were due to the composition of the juice (types of fruit) and the storage or processing conditions. The vitamin C content of red pitaya is within the same range as other common fruits such as apple (6 mg/100 g) and plum (3 mg/100 g) as reported by Belitz and Grosch (1999). A study done by Miller and Rice-Evans (1996) suggested that the phenolic antioxidants of fruit juices protect the vitamin C content from oxidative degradation. Phenolics are important because of their contribution to the sensory quality of fruits (color, astringency, bitterness and avor), which may be affected during the technological processes used for obtaining the juices and other transformation products (Ramadan and Moersel 2007). Fair amounts of polyphenols were found in red pitaya juice that were 1.2441.336 mg/100 g juice as gallic acid equivalents. Higher amounts of phenolics were found in processed pitaya juice as shown by the chart in Fig. 3 , which suggests that pitaya beverages are rich in antioxidants capacity than in the unprocessed mash. The presence of phenolics components play a signicant contributory role as antioxidant; which is important in the maintenance of health and protection from coronary heart disease and certain cancers (Miller and RiceEvans 1996). CONCLUSIONS Recently, pitayas have attracted great interest from the fruit growers to the fruit processing industries due to their nutritional and health benets. The

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utilization of dragon fruit or pitaya has also been widened to produce wine, soft drinks, jam, fruit concentrate and enzyme drink. However, the dragon fruit products such as the dragon fruit juice concentrate that has been marketed in small and medium scale industry in Malaysia does not have good quality in terms of structure and appearance. Hence, processing of dragon fruit juice using enzyme is important to produce high quality fruit juice that meets the consumer interest. Enzyme such as pectinase has been used since a long time ago as a processing aid in fruit juice production. Pectinase breaks the structure of plant cell wall and releases the micro-and macro-components contained therein. Application of enzyme proved to increase the juice recovery as well as the juice physicochemical characteristics. It also increased the nutritional composition in the juice such as protein and total polyphenols. The protein content after enzymatic treatment using Pectinex CLEAR increased to 2.20% w/w from 0.23% w/w of the raw juice. Total polyphenols content also increased by 7% from its initial value after treatment using Pectinex Ultra SP-L enzyme. The differences in chemical compositions, physicochemical parameters, vitamin C and total polyphenols between untreated juice when compared to enzyme-treated juice show that the mechanism of pectin degradation depends upon the types of enzymes used. From the study done, it can be seen that the effects of Pectinex Ultra SP-L and Pectinex CLEAR were different on red pitaya juice because pectinases act in different ways on the pectins. Pectins are situated in the primary cell walls and in the middle lamella. The mechanism of pectin degradation depends upon the pectic composition of the substrate. This study demonstrated the effect of pectinases application on the chemical composition, vitamin C and total polyphenols of red pitaya juice. Moreover, this study aims to obtain some data from the processing of fruit juice especially pitaya juice and to provide valuable and interesting information that would be useful for fruit processing industries. Utilization of pectinase as a processing aid in the production of pitaya juice proved to be a crucial step in order to enhance the efciency of the whole system.

ACKNOWLEDGMENTS The authors would like to thank Ms. Rossdiannawatty Hassan and Miss Nurul Hida for their excellent assistance in the production of pitaya juice. We also would like to acknowledge staffs of Biochemistry Laboratory (Faculty of Food Science & Technology, UPM) and Mr. Badrushah (Department of Process and Food Engineering, UPM) for all the supports given.

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