Rapid Extraction of Pawpaw Juice with the Application of Locally Produced Pectic Enzymes from Saccharomyces Cerevisiae ATCC 51712

Pawpaw (Carica papaya) fruit is highly perishable; hence, conversion into a value-added product such as juice concentrate will be useful. Pawpaw fruit is, however, high in pectin; therefore, its juice extraction is difficult without an extraction aid such as the addition of pectic enzymes. Our laboratory has developed a simple enzymatic method that enables the extraction of pawpaw juice with considerable ease. Pectic enzymes were obtained from Saccharomyces cerevisiae (ATCC 52712) cultured in pawpaw juice for 6 days at room temperature (25–28°C) and centrifuged to obtain the enzyme in the supernatant. Addition of this enzyme (32 mg total protein) to 200 g pawpaw mash and incubation for 30 min resulted in significant (P < 0.05) reduction in the viscosity of the mash with corresponding increases in the flow rate of free-run juice (85%). Simultaneously, large increases in the volume of free-run juice (180%) over the controls were obtained. Since the overall increase in total juice volume was only 35% as compared with 180% for volume of free–run juice and 80% increase in flow rate for free–run juice, it was concluded that the major effect of the enzyme treatment on pawpaw juice extraction is on the decrease in processing time as a result of the degradation of the pectin in the fruit flesh.     

Enzymatic Clarification of Fruit Juices by Fungal Pectin Lyase

The activity of pectic enzymes was measured in culture filtrates of four selected fungal strains. Pectin lyase (PL) activity was produced by all fungi, but pectinesterase activity was not detected in culture filtrates of the strain Penicillium expansum F16. The use of crude enzymes resulted in good clarification of apple juice as evaluated by an increase in transmittance at 660 nm. PL activities were partially purified by dye-affinity chromatography on Blue Sepharose. The enzyme was bound to the column and eluted in a single peak, free of PE activity, and not coinciding with the major amount of protein. Apple, grape, and passion fruit juices were effectively clarified by these partially purified enzymes. Methanol content was determined by gas-liquid chromatography, being undetectable in the clarified juices. These pectic enzymes demonstrate potential for use in fruit juice processing.     

EFFECT OF PECTINOLYTIC AND AMYLOLYTIC ENZYMES ON APPLE JUICE TURBIDITY

The mechanisms governing the enzymatic clarification of apple juice were studied by electron microscopy techniques. Full ripe and unripe apple juice samples (Granny Smith) were treated with commercial pectinase (Solvay 5XLHA) and amylase (Röhalase HT) enzymes, respectively. Scanning electron microscopy studies revealed that commercial amylolytic enzymes quickly reduced starch content in unripe apple juice to undetectable values. It was also observed that after pasteurization of this juice (90C, 5 min) all starch granules gelatinized. Using transmission electron microscopy, it was possible to observe pectin bonded to ripe apple juice particles. This protective colloid is known to be responsible for cloudy juice stability. The effect of pectic enzyme to destroy the protective pectin colloid was also detected with this technique. As a result of the enzymatic treatment, average particle size initially increased from 1000 to 1500 nm and decreased thereafter to ≈1100 nm, and Z‐potential increased in absolute values from ?9.6 to ?11.4 mV. It was speculated that the destruction of the weak pectin net by the action of the specific enzyme caused particle aggregation, followed by the collapse of aggregates, increasing the number of particles <500 nm.     

THE EFFECT OF PROCESSING METHOD ON THE CHARACTERISTICS OF CARROT JUICE

ABSTRACT

The production of carrot juice using different technologies is the main subject of this investigation. Although the production and preservation of the carrot juice technology have not been improved and are not prevalent, an intensive study has been carried out on this subject recently. In this study, the effects on the yield and quality of carrot juice produced with and without total enzymatic liquefaction (Pectinex Ultra SP‐L, Novo Nordisk Ferment, Dittingen, Switzerland) as well as lactofermentation or citric acid addition have been investigated. Generally, enzymatic liquefaction of mash increased the juice yield (89.32 ± 1.17 and 90.43 ± 0.47% for lactofermented and citric‐acid‐added samples, respectively), while the control sample juice yield was found to be 59.90 ± 0.81%. Lactofermented and acid‐added samples demonstrated lower pH value. In enzyme‐treated samples, the total acidity increments were also determined. Accordingly, galacturonic acid formed after enzymatic breakdown of pectin in the raw material increased the total acidity of the samples. Magnesium contents of lactofermented juices were found to be low because lactic acid bacteria require magnesium for their growth. β‐Carotene contents have been found to be higher in both lactofermented and in nonenzyme‐treated samples.

PRACTICAL APPLICATIONS

This article describes the effects of different processing methods on the characteristics of carrot juice. It is difficult to preserve vegetable juices such as carrot juice because of their low acidity. The acidity of carrot juice (pH ～6.2) can be adjusted by the addition of citric acid or the lactofermentation method to pH 4.5. With these processes carrot juice can be preserved by pasteurization as occurs for low pH fruit juices, increasing shelf life. According to the results of this study, increasing the acidity of mash with lactofermentation or citric acid addition also increases the juice yield.

     

桑果醋发酵工艺研究

以桑果、糯米为原料，采用液态发酵法，酿制风味独特、营养丰富的桑果醋。通过对桑果汁压榨、浸提、澄清和酒精发酵、醋酸发酵等工艺的研究，确定桑果汁酶解澄清的最佳方案是：果胶酶添加量为0．035％、淀粉酶0．014%、明胶0．018％、硅溶胶0．018％；确定主要工艺参数为：酒精发酵阶段桑果清汁：糯米糖化醪=2：1，接种量5％，25℃发酵72h，酒精浓度6．0％，醋酸发酵阶段温度为33℃-35℃，接种量6．0％，起始pH6．0，500mL三角瓶装液量100mL，在转速140r／min条件下振荡培养72h左右，得到的成品醋酸度适宜，口感、风味良好。     

PRODUCTION AND USE OF YEAST PECTOLYTIC ENZYMES TO AID PINEAPPLE JUICE EXTRACTION

A crude preparation of pectolytic enzyme was obtained by culturing Saccharomyces cerevisiae (ATCC512712) in pineapple juice. Maximum cell growth was obtained on the 6th day after inoculation. Correspondingly, optimum protein concentration and enzyme activity were also observed on the 6th day old culture. Although not purified, the addition of this crude enzyme extract to crushed pineapple mash at 0.02% resulted in a significant increase in juice yield (P≥0.05). The overall processing time was also significantly reduced (P≤0.05) with the enzyme treatment compared to controls. A combination of 0.02% enzyme treatment of mash for 30 min, was found to be the most effective dosage in reducing the processing time significantly.     

THE EFFECTS OF DIFFERENT INITIAL LACTOBACILLUS PLANTARUM CONCENTRATIONS ON SOME PROPERTIES OF FERMENTED CARROT JUICE

Carrot juice (Daucus carota L.) is one of the most popular vegetable juices and represents a rich source of natural β‐carotene. Vegetable juices are available either fermented or not fermented. With the production of lactofermented juices, carrot juices are microbiologically stable, delicious and potentially provide high nutritional value. In this study, Refik Saydam Kulfur Kolleksiyanu (RSKK) 1602 Lactobacillus plantarumis used as a starter culture in carrot mash. Sensory evaluation demonstrated that an initial bacteria concentration of 3 × 10⁵ cfu/g mash resulted in the most preferred fermented carrot juice. The acidity of the fermented carrot juice can be adjusted by altering the population of the starter RSKK 1602 L. plantarum culture and the 15–16 h of fermentation time. A modified Gompertz equation was selected to describe the growth curve of RSKK 1602 L. plantarum in carrot mash.     

Enzymatic Clarification of Fruit Juices by Fungal Pectin Lyase

The activity of pectic enzymes was measured in culture filtrates of four selected fungal strains. Pectin lyase (PL) activity was produced by all fungi, but pectinesterase activity was not detected in culture filtrates of the strain Penicillium expansum F16. The use of crude enzymes resulted in good clarification of apple juice as evaluated by an increase in transmittance at 660 nm. PL activities were partially purified by dye-affinity chromatography on Blue Sepharose. The enzyme was bound to the column and eluted in a single peak, free of PE activity, and not coinciding with the major amount of protein. Apple, grape, and passion fruit juices were effectively clarified by these partially purified enzymes. Methanol content was determined by gas-liquid chromatography, being undetectable in the clarified juices. These pectic enzymes demonstrate potential for use in fruit juice processing.     

Effect of macerate enzymes on the yield,quality, volatile compounds and rheological property of prickly pear juice

Pectinase and cellulase enzymes were used to investigate efficacy for improving juice yield, stability and quality from prickly pear fruit. Pectinase improved the yield, stable color, color‐assayed as release of anthocyanins or carotinoids and clarity of the juice. A significant increase in the effectiveness of pectinase was observed as the concentration was increased from 0.05 to 0.50% v/w. However, at concentration >0.25% v/w they tended to impart a bitter flavor in the juice. Among three concentrations of pectinase and cellulase, pectinase at 0.50% v/w produced higher yield, a sediment‐free clear juice and high‐quality juice. The results indicated that depectinated clarified prickly pear juice behaves as a Newtonian fluid. It was found that the activation energy (Ea) for viscous flow was in the range of 5.02×10³–20.06×10³ kJ/mol depending on the concentrations of pectinase and cellulase enzyme treatment of prickly pear juice, in contrast to 22.15×10³ kJ/mol in untreated juice. Volatile compound concentrations of twelve compounds were not affected by pectinase and cellulase treatment. Overall the quality of prickly pear juice was better in pectinase‐treated juice compared with untreated and cellulase‐treated juice.     

Ethephon‐induced fractional changes of pectic polysaccharides in developing cape gooseberry (Physalis peruviana L) fruits

Pectic polysaccharides extracted from fruit tissue of cape gooseberry (Physalis peruviana L) at different maturity levels were fractionated and analyzed for galacturonic acid backbone molecules. Preharvest spray of the fruits at 7 days old with ethephon resulted in a higher accumulation of water‐soluble and oxalate‐soluble pectic fractions than non‐treated fruits, while this treatment effectively enhanced the polygalacturonase enzyme activity necessary for maintenance of pectin degradation. Treatment with ethephon not only enhanced the solubilization rate but also resulted in a net increase in total pectic polymer content in developing cape gooseberry fruits. Copyright © 2005 Society of Chemical Industry     

DEVELOPMENT OF A MASHING PROFILE FOR THE USE OF MICROBIAL ENZYMES IN BREWING WITH RAW SORGHUM (80%) AND MALTED BARLEY OR SORGHUM MALT (20%)

Different time and temperature programmes were used to evaluate the production of hot water extract (HWE) and free amino nitrogen (FAN) from mashes containing raw sorghum and either malted sorghum or malted barley in the presence of microbial enzymes. Two malted varieties of sorghum (SK 5912 and Zaria) were used. The former gave higher HWE but lower FAN than the latter. Sorghum malts were unable to provide enzyme activity for starch extraction and exogenous enzymes were always needed. Seventeen commercially available enzyme preparations were assessed. A double-mash process was developed. Inclusion of calcium ions (200 ppm) was beneficial but adjustment of mash pH had little effect. Raw sorghum was gelatinised at 100°C for 30–40 min in the presence of a heat-stable α-amylase followed by mixing with a malt mash (started at B0°C) to give a temperature of 65°C with a total mash time of 167 min (127 min from mixing the mashes). The inclusion of a single commercial enzyme preparation (containing both proteolytic and amylolytic activities) was sufficient to achieve satisfactory HWE and FAN. Addition of different activities or combinations of activities gave no significant advantages. To obtain levels of FAN of 100–140 mg/l however excessive amounts of enzymes were required.     

REDUCTION OF SACCHAROMYCES CEREVISIAE, ESCHERICHIA COLI AND LISTERIA INNOCUA IN APPLE JUICE BY ULTRAVIOLET LIGHT

Apple juice was inoculated separately with Saccharomyces cerevisiae, Listeria innocua (ATCC 51742) or Escherichia coli (ATCC 11775) for treatment in a double tube ultraviolet (UV) disinfection system. The apple juice was treated at six flow rates (0.073–0.548 L/min) for selected fluences (75–450 kJ/m²). The juice was also inoculated with a mixture of these three microorganisms and UV light treated from 0.548 to 0.735 L/min for 30 min. The microbial reduction was described with a first order kinetic model. Average D_uvvalues of 23.1–40.5, 8.2–20.6 and 6.0–17.7 min were obtained for S. cerevisiae, L. innocua and E. coli, respectively. A linear model was used to describe the relationship between log D_uvversus flow rate for S. cerevisiae and L. innocua. However, a third order polynomial model was more adequate for describing the E. coli D_uvvalues versus flow rate. Less than 10 (no growth), 190 and 200 cfu/mL of S. cerevisiae, L. inocua and E. coli, respectively, were observed in UV‐treated apple juice inoculated with a mixture of microorganisms.     

Suitability of Bifidobacterium spp. and Lactobacillus plantarum as Probiotics Intended for Fruit Juices Containing Citrus Extracts

A strain of Lactobacillus plantarum and 4 strains of bifidobacteria were inoculated in apple juice and in a commercial beverage labeled as “red‐fruit juice,” containing citrus extracts as natural preservatives; the suitability of the probiotics was evaluated in relation to their resistance to 2 kinds of citrus extracts (biocitro and lemon extract), survival in juices at 4 and 37 °C, and inhibition of Zygosaccharomyces bailii. Cell count of L. plantarum and bifidobacteria over time was fitted through the Weibull equation, for the evaluation of the first reduction time (δ), death time, and microbiological shelf life (the break‐point was set to 7 log cfu/mL). Bifidobacterium animalis subsp. lactis experienced the highest δ‐value (23.21 d) and death time (96.59 d) in the red‐fruit juice at 4 °C, whereas L. plantarum was the most promising strain in apple juice at 37 °C. Biocitro and lemon extract did not exert a biocidal effect toward probiotics; moreover, the probiotics controlled the growth of Z. bailii and the combination of L. plantarum with 40 ppm of biocitro reduced the level of the yeast after 18 d by 2 log cfu/mL.     

Effects of different enzymatic maceration treatments on enhancement of anthocyanins and other phenolics in black currant juice

Pre-press maceration treatments with 10 different pectinolytic enzyme preparations were investigated in experimental black currant juice production using response surface design templates. Enzyme dosage, maceration time, reaction temperature, and degree of berry crushing were varied, and the juice yields, anthocyanins, total phenols, and turbidity in the resulting juices were compared for a total of 250 different enzymatic treatments. The yields of anthocyanins and total phenols in the juices ranged from 900 to 2200 and 3050 to 5100 mg/kg wet weight black currant mash, respectively. Juice yields ranged from 66.4% to 78.9% by wet weight of mash. Turbidity levels ranged from 25 to 916 formazan nephelometric units (FNU). The reaction parameters induced larger variations in the responses than the different enzyme preparations, but the cloned Aspergillus niger/Aspergillus aculeatus preparation Pectinex BE® consistently tended to be among those giving the best responses regarding anthocyanin yields, phenols, and low juice turbidity. The optimal maceration was achieved using an enzyme dosage of 0.18% by wet weight of berries with a reaction at 60 °C for 30 min on the most finely crushed berry mash. This treatment gave similar profiles of anthocyanins in the juices with all the 10 enzyme preparations. The same 10 juices all exhibited antioxidant activity against human low-density lipoprotein oxidation in vitro, but the antioxidant potency varied depending on the enzyme preparation used in the pre-press maceration.     

Effect of juice processing on the cancer chemopreventive effect of cranberry

Cancer chemopreventive properties were evaluated in cranberries and cranberry products (mash, depectinized mash, pomace, raw juice, clarified juice and juice concentrate). Three extracts isolated from frozen cranberries and cranberry solids (mash, depectinized mash and pomace) containing anthocyanins, water-soluble and apolar phenolic compounds were tested. Cranberry juices and extracts were screened for their ability to induce the phase II xenobiotic detoxification enzyme quinone reductase (QR). The results showed that there was no cytotoxicity against the cells used in the test. All samples stimulated quinone reductase activity except the highest concentrations of the anthocyanin-rich extract of pomace, which inhibited QR activity. Also, the results showed that the QR induction for all samples varied with concentration and that there was an optimal concentration for which the QR induction was maximal. Although the three cranberry extracts were good QR inducers, our results indicated that the phenols present in aqueous extract showed QR inductions which were more important than those obtained with phenols present in solvent extracts. Also, the ability of phenols to stimulate the QR activity has been reduced continuously and significantly (P ≤ 0.05) during the technological process. Especially, it appears that conditions of the evaporation to obtain a juice concentrate exerted a significant effect (P ≤ 0.05) on inducer potencies of bioactive molecules.     

Oil extractability from enzymatically treated goldenberry (Physalis peruviana L.) pomace: range of operational variables

Goldenberry pomace (seeds and skins) represents a large portion of the waste generated during juice processing (ca. 27.4% of fruit weight). The potential of goldenberry agro‐industrial wastes for use as raw material for production of edible oil was evaluated. Fruit pomace, contained 6.6% moisture, 17.8% protein, 3.10% ash, 28.7% crude fibre and 24.5% carbohydrates. The n‐hexane‐extractable oil content of the raw by‐products was estimated to be 19.3%. Aqueous enzymatic extraction was investigated for recovery of oil from the fruit pomace. The most significant factors affecting extraction were enzyme concentration, the time of digestion with enzymes, substrate concentration in water and the particle size of substrate. A broad variation in oil recovery was obtained depending on the operational conditions during the enzyme‐aided aqueous extraction. The optimum and economical values were those obtained for 4:0.02:1 water:enzyme:substrate ratio. Generally, enzymatic treatment increased the extraction yield. The more than 42% yield by enzymation compared with the nearly 3% yield in the control process (without enzyme) implies a significant increase in yield by about 92.8%. In single‐enzyme trials, cellulase EC gave the best yield. Although proteases slightly improve yield, the enhancement values are much lower than those obtained with Cellulase EC and Pektinace L40. Rapid increase in oil yield occurred as the enzyme concentration increased from 1 to 2 g/100 g substrate. Yield increased with dilution, but it began to fall when the substrate became more diluted. Moreover, extractability increased significantly when particle size reduced. Concerning the oil composition, there were no great changes in the fatty acid pattern of the oils extracted with different hydrolytic enzymes when compared with each other or to the solvent extracted oil. The main purpose of this study was to maximise the efficiency of the enzymatic treatment for oil recovery from goldenberry pomace. As a first step toward developing goldenberry as a commercial crop, the results provide important information for the industrial application of the fruit.     

Effects of pulsed electric field treatment of apple mash on juice yield and quality attributes of apple juices

The influence of apple mash treatment with pulsed electric fields (PEF) on yield and quality attributes of the resulting cloudy juices were investigated. Apple juice was produced at laboratory scale (400 g apples per lot) by mash treatment with pulsed electric fields at three different field intensities (1, 3, 5 kV/cm, n = 30 pulses) and manual pressing at room temperature. The juices were compared with untreated control juices of the same mash and with a juice after pectolytic mash treatment. Relative to the control samples, juice yield increased with increasing field intensities. The overall composition as described by pH, total soluble solids, total acidity, density, contents of sugar, malic acid and pectin, respectively, as well as the nutritive value with respect to polyphenol contents and antioxidant capacities (TEAC, FRAP, DPPH) of the PEF-treated apple juices did not significantly differ from the controls.Industrial relevanceThis paper addresses the composition of apple juice after mash treatment applying pulsed electric fields and maceration enzymes, respectively. Since substantial equivalence of products derived from novel technologies, such as PEF, relative to their conventionally produced counterparts is required according to European food law, this aspect is of great relevance, especially for commercialisation of fruit juices based on PEF technology. Furthermore, the nutritional value of juices produced at different pulsed electric field activities is of relevance for their marketing.     

Effect of chitosan coating in preventing deterioration and preserving the quality of fresh‐cut papaya ‘Maradol’

BACKGROUND: Chitosan can form antimicrobial, semi‐permeable barriers that limit gas exchange and reduces water loss in fruits. Consumer interest in fresh‐cut papaya fruit is leading to increasing demand because of its sensorial and antioxidant properties. However, papaya is a highly perishable product that is prone to loss of weight, loss of firmness and microbial attack. The aim of this study was to evaluate the effect of chitosan coatings on the overall quality of fresh‐cut papaya. Chitosan coatings of low (LMWC), medium (MMWC) and high (HMWC) molecular weights, at concentrations of 0.01 and 0.02 g mL^?1, were applied to fresh‐cut papaya cubes. The treated cubes were stored at 5 °C and changes in quality were evaluated. RESULTS: MMWC maintained the highest color values (L* and b*) and firmness. Chitosan coatings suppressed mesophilic plate count, and the growth of molds and yeast, compared to controls. The MMWC coatings at 0.02 g mL^?1 resulted in the highest antimicrobial activity and decreased the activity of the enzymes polygalacturonase and pectin methylesterase, followed by low and high MW chitosan coatings at 0.02 g mL^?1. CONCLUSION: The application of the MMWC treatment at 0.02 g mL^?1 could be used to reduce deteriorative processes, maintain quality and increase the shelf life of fresh‐cut papaya stored at 5 °C. Copyright © 2008 Society of Chemical Industry     

THE USE OF PULSED ELECTRIC FIELDS IN PRODUCING JUICE FROM PAPRIKA (CAPSICUM ANNUUM L.)

Investigations were conducted on the impact of pulsed electric field (PEF) treatment on yield and some quality parameters of juice from paprika and results were compared to the results of juice obtained from enzymatically treated or untreated paprika mash. Paprika was comminuted mechanically after which the mash was subjected to either PEF (electric field strength = 1.7 kV/cm, pulse number = 30 and specific energy input per pulse = 0.5 kJ/kg) or a pectolytic enzyme preparation treatment. The extent of cell disintegration after the different pretreatments was monitored through cell disintegration index (Z_p) measurement prior to juice extraction through pressure (10 MPa for 4 min). Some quality parameters (pH, soluble solids (°Brix), total dry matter, color, total carotenoids as β-carotene and vitamin C) of the resultant juice samples were determined. PEF and enzyme treatments resulted in about 10% and 9% increase in juice yield, respectively. Juice from PEF treated paprika compared well in quality with enzyme treated or the untreated. And in some cases such as in color (redness, a value) of the juice, PEF treated had values (+ 18) higher than enzyme treated having + 15.5 and also in the amount of β-carotene extracted into the juice PEF treated juice had more than 60% as compared to about 44% from enzyme treated juice. The pH value for enzyme treated juice was lower than the others.     

Purification and characterization of an amylase from Opuntiaficus‐indica seeds

BACKGROUND: In Tunisia, prickly pear fruit grow spontaneously; it is consumed as fresh fruit, juice or jam. When the fruit is used for juice production, the seeds are discarded and go to waste. Our study aimed to extract biomolecules from seeds by producing value‐added products from the fruits. RESULTS: An amylase from Opuntia ficus‐indica seeds was extracted and purified to homogeneity. An increase in specific activity of 113‐fold was observed. The apparent molecular mass of the enzyme is 64 kDa. The optimum pH and temperature for enzyme activity were pH 5 and 60 °C, respectively. Under these conditions, the specific activity is 245.5 U mg^?1. The enzyme was activated by Co²⁺ and Mg²⁺ (relative activity 117% and 113% respectively) at lower ion concentrations. It was strongly inhibited by Mn²⁺ and Fe²⁺. Cu²⁺ inhibited totally the activity of this enzyme, but Ca²⁺ has an inhibitory effect which increases with ion concentration. CONCLUSION: The extracted enzyme belongs to the exo type of amylases and is classified as a β‐cyclodextrin glycosyltransferase since it generates mainly β‐cyclodextrin from starch. It exhibits high thermal stability and a broad range of pH stability, making it a promising prospect for industrial and food applications. Copyright © 2012 Society of Chemical Industry