Fruit Juice Clarification Research Articles

Valorization of agro-industrial residues for pectinase production by Aspergillus aculeatus: Application in cashew fruit juice clarification

Valorization of agro-industrial residues for pectinase production by Aspergillus aculeatus: Application in cashew fruit juice clarification

Characterization of a New Laccase from Vibrio sp. with pH-stability, Salt-tolerance, and Decolorization Ability.

Laccases have been widely used for fruit juice clarification, food modification, and paper pulp delignification. In addition, laccases exhibit remarkable performance in the degradation of toxic substances, including pesticides, organic synthetic dyes, antibiotics, and organic pollutants. Thus, the screening and development of robust laccases has attracted significant attention. In this study, Vibrio sp. LA is a strain capable of producing cold-adapted laccases. The laccase coding gene L01 was cloned from this strain and expressed in Yarrowia lipolytica, a host with good secretion ability. The secreted L01 (approximate MW of 56,000 Da) had the activity and specific activity of 18.6 U/mL and 98.6 U/mg toward ABTS, respectively. The highest activity occurred at 35 °C. At 20 °C, L01 activity was over 70% of the maximum activity in pH conditions ranging from 4.5-10.0. Several synthetic dyes were efficiently degraded by L01. Owing to its robustness, salt tolerance, and pH stability, L01 is a promising catalytic tool for potential industrial applications.

Clarification of citrus fruit juices using microfiltration technique equipped with indigenously developed novel ceramic membrane.

The online version contains supplementary material available at 10.1007/s13197-023-05734-y.

Characterization of Two Novel Rumen-Derived Exo-Polygalacturonases: Catalysis and Molecular Simulations

Pectinases are a series of enzymes that degrade pectin and have been used extensively in the food, feed, and textile industries. The ruminant animal microbiome is an excellent source for mining novel pectinases. Two polygalacturonase genes, IDSPga28-4 and IDSPga28-16, from rumen fluid cDNA, were cloned and heterologously expressed. Recombinant IDSPGA28-4 and IDSPGA28-16 were stable from pH 4.0 to 6.0, with activities of 31.2 ± 1.5 and 330.4 ± 12.4 U/mg, respectively, against polygalacturonic acid. Hydrolysis product analysis and molecular dynamics simulation revealed that IDSPGA28-4 was a typical processive exo-polygalacturonase and cleaved galacturonic acid monomers from polygalacturonic acid. IDSPGA28-16 cleaved galacturonic acid only from substrates with a degree of polymerization greater than two, suggesting a unique mode of action. IDSPGA28-4 increased the light transmittance of grape juice from 1.6 to 36.3%, and IDSPGA28-16 increased the light transmittance of apple juice from 1.9 to 60.6%, indicating potential application in the beverage industry, particularly for fruit juice clarification.

Low-Fouling Plate-and-Frame Ultrafiltration for Juice Clarification: Part 1—Membrane Preparation and Characterization

Membrane technology provides advantages for separating and purifying food materials, including juice clarification. Ultrafiltration processes for fruit juices aim to remove haze components and maintain the stabilization of the juices. However, the membrane process during the clarification of fruit juices suffers from fouling, which deteriorates the process performance and products. Biofouling usually is found in the applications of the membrane in food processing. In this study, phenolic substances extracted from garlic bulbs are incorporated into a matrix of polymeric membranes to prevent fouling during juice clarification. Hydrophilic cellulose acetate was used as the polymer matrix of the membrane, and dimethylformamide was used as the solvent. The phenolic substances from garlic bulbs were incorporated into polymer solutions with different concentrations of 0%, 1%, 1.25%, and 1.5% w/v. The composite membrane was prepared using the phase inversion method, and the resulting membranes were later characterized. The results show that incorporating those phenolic compounds as the dope solution additive resulted in membranes with higher hydrophilic properties. They also modeled antibacterial properties, as shown by the reduced attachment of Bacillus subtilis of up to 91.5 ± 11.7% and Escherichia coli of up to 94.0 ± 11.9%.

Extracellular pectinase production from a novel Yarrowia phangngaensis XB3 grown on banana waste and its application in fruit juice clarification

Pectinases are important enzymes used in several industries, including food and beverage. Yarrowia phangngaensis, a yeast with pectinase-producing potentials, was investigated for its ability to produce copious pectinase using agro-wastes as substrates. Culture optimization studies were conducted, followed by partial purification by ethanol precipitation and application in apple juice clarification. The optimum pH and temperature for the pectinase activity were studied, and the suitable parameters for apple juice clarification were determined. Culture optimization remarkably influenced pectinase production by Yarrowia phangngaensis. Banana peels elicited the highest pectinase production compared to potato, pawpaw, orange and lemon peels. Yeast extract and ammonium sulphate proved best for pectinase synthesis. The optimum condition for pectinase production was 5% banana peel, pH 7.5, 1.4 × 104 cells ml−1 and incubation for 36 h. Culture optimization increased the pectinase yield by 182%. Ethanol precipitation (50%) resulted in a 5.31-fold increase in the specific activity of the pectinase. The optimum juice clarification was achieved by treating 10 ml apple juice with 7.03 IU ml−1 pectinase for 120 min. The turbidity and clarity reduction percentages were 39.33% and 59.84%, respectively. The pectinase from Yarrowia phangngaensis is applicable in food and other industries.

Pectinase Production by Aspergillus niger and Its Applications in Fruit Juice Clarification

Pectinases (EC 3.2.1.15) are a class of enzymes that catalyze the depolymerization or de-esterification reactions that degrade pectic substances. In the present study, we have isolated the Aspergillus niger strain from soil samples, in India and evaluated pectinase production. The highest pectinase producing A. niger strain was further evaluated and optimized with various agricultural wastes. Plackett-Burman design (PBD) and Central composite design ‘(CCD)’ were used to determine the best parameters for maximum pectinase production. Pectinase activity was increased to 99.21 U/ml after optimizing the production medium using PBD and CCD statistical analysis. A positive correlation of pectinase activity between predicted (112.65 U/ml) and experimental (99.21 U/ml with SD=0.005) optimum was observed. Maximum pectinase was produced by A. niger under submerged fermentation, utilizing orange peel, which is a cost-effective, adaptable, and environmentally friendly approach. The partially purified pectinase showed significant application for apple juice clarification and showed the ability to degrade pectin and therefore the colour change was observed in apple juice within 120 min. Maximum pectinase was produced by A. niger using agricultural waste orange peel under submerged fermentation which is an economical, versatile and eco-friendly process and pectinase showed a significant application for apple juice clarification.

CLARIFICATION TREATMENTS OF PINEAPPLE (Malus domestica) AND ORANGE (Citrus sinensis) JUICES BY PECTINASE FROM Yarrowia lipolytica STRAINS IDENTIFIED FROM COCOA JUICE IN FERMENTATION

Background: Pectinases are a group of enzymes that break down pectin, a polysaccharide that is found in plant cell walls. Today, the application of pectinolytic enzymes plays an important role in food technology for the maceration of fruits and vegetables, including for the extraction and clarification of juice.
 Aim of the Study: This study aimed to test the clarifying ability of pectinase of three Yarrowia lipolytica strains (Tias J0-6; YA J3-1; Buy J2-1) identified for cocoa juice in fermentation.
 Materials and Methods: The pectinase from Yarrowia lipolytica strains (Tias J0-6; YA J3-1; Buy J2-1) were produced using a basic liquid medium and partially purified through ion exchange chromatography. The fruit juices were clarified by enzymatic treatments against a control (free enzymatic treatment juice) and physicochemical and phytochemical parameters and antioxidant activities of the clarified juices were determined.
 Results: The results revealed that from three strains, Tias J0-6 strain produced pectinase which showed the best clarification activity on pineapple and orange juices. The juices clarified exhibited an increasing of clarity values and decreasing viscosity and pH values. Thus, pectinase of Tias J0-6 allowed increasing clarity to 41.81% and 7.27%, to decrease viscosity up to 14.03% and 6.55% respectively for pineapple juices and orange juices clarified. The pH values decreased from 4.036±00 to 4.031±00 and from 5.010±00 to 4.592±00 respectively for orange and pineapple juices. The antioxidant activities of juices clarified were strongly correlated to total tannins and total flavonoids (r = 0.717, r = 0.754, r = 0.855).
 Conclusion: Tias J0-6 strain seemed a suitable candidate to biotechnology applications such as fruit juices clarification.

Engineering magnetic nanobiocatalytic systems with multipurpose functionalities for biocatalysis, biotechnology and bioprocess applications

Enzymes are high-performance natural biological catalysts with wide-ranging applications in agricultural, medical, food and environmental sectors. Nevertheless, lack of efficient recovery, reusability, and high cost of the soluble form of enzymes are the most daunting challenges rendering biocatalytic systems inadequate for industrial exploitation. In order to deal with these inadequacies, immobilization appears to be a prodigious approach for enhancing the stability and catalytic efficiency of enzymes, as well as enabling their separation and reusability in continuous reaction batches. Among different nanostructures, magnetic nanomaterials have garnered supreme interest as support matrices for biomolecules and enzymes immobilization because of their substantial surface area, larger surface-to-volume ratio, modifiable surface, and adjustable surface particle size, stability, and high mass transferring ability. In addition, they can be quickly recovered from the complex reaction system by a simple external magnetic field. Magnetic nanoparticles incorporated biocatalysts demonstrated a broad-working temperature and pH profile and augmented storage and thermal stabilities compared to their native derivatives. This paper provides a recent and state-of-the-art overview of the development and application of multifunctional magnetic nanobiocatalytic systems for an array of biotechnological purposes. In the first half, the development, functionalization, and use of nanostructured magnetic materials as enzyme immobilization supports are delineated. Then the prospective applications of magnetic nanobiocatalytic systems in different industrial sectors, including wastewater treatment, biodiesel and butanol production, hydrolysis of lignocellulosic biomass, glucose monitoring, fruit juice extraction and clarification, and synthesis of non-natural benzylisoquinoline alkaloid are comprehensively vetted with representative examples. Finally, the conclusive points and future prospects in this evolving field are also directed.

Candida krusei is the major contaminant of ultrafiltration and reverse osmosis membranes used for cranberry juice production

Ultrafiltration (UF) and reverse osmosis (RO) are commonly used for the clarification and concentration of fruit juices. However, one of the main limitations of filtration membranes is biofouling, which reduces membrane efficiency and can contaminate the filtered product and lead to spoilage. In this study, the microbial fouling layers of UF and RO membranes from a Canadian cranberry juice processing plant were characterized. Unlike the microbiota found in cranberry juice, which is dominated by Bacillus sp. and other bacteria, both UF and RO membranes were mainly colonized by several strains of the yeast Candida krusei. A variation in bacterial and yeasts count was observed between tubular UF and spiral-wound RO membranes, and the analysis of the spatial distribution highlighted the homogeneity of the contamination across each membrane. Surprisingly, RO membranes had a higher level of contamination when compared to UF membranes. Furthermore, six strains of C. krusei were further characterized through multilocus sequence typing analysis, five of which exhibited unique allelic profiles and two of which were found to contain a new TRP1 allele.

Production of microalgae-derived exo-polygalacturonase for fruit juice clarification

Due to the growing demand for commercial enzymes derived from renewable and sustainable resources with higher efficacy, algal biomass has gained more attention. Thus, the production of exo-polygalacturonase (PGase) from Bacillus subtilis under submerged fermentation (SmF) using algal feedstocks as a carbon source in fermentation medium and its potential use for the clarification of apple and carrot juices with the optimization of the enzyme loading, pH, and time using Box–Behnken response surface methodology (RSM) were prompted. The maximum PGase activity of 510.61 ± 7.86 U/L was achieved at pH 5.0 after 48 h at 40°C using 0.5% (w/v) pectin, 0.1% (w/v) yeast extract, 0.02% (w/v) MgSO47H2O, 0.02% (w/v) K2HPO4, 0.3% (w/v) glucose, and 0.583% (w/v) microalgae hydrolysate as a fermentation medium. PGase production without glucose was favored by SmF with a shake flask working volume of 100 ml (248.90 ± 4.58 U/L) as compared to a bioreactor with a working volume of 1 L (192.99 ± 0.84) for 48 h. Both carrot and apple juices were treated with the crude PGase (248.9 U/L) at different concentrations (0.1–0.5%), temperature (30–50°C), and time (30–120 min), and the maximal clarification conditions were obtained as 0.3% (w/v) enzyme concentration at 50°C for 30 min exposure time. This study revealed that microalgae-derived PGase in crude form could be effectively used for clarification of the juices.

Isolation and characterization of pectinase-producing bacteria (Serratia marcescens) from avocado peel waste for juice clarification

BackgroundBacterial pectinase is an enzyme that could be employed in numerous sectors to break down pectin polysaccharide compounds. The goal of this study is to find pectinase-producing bacteria in avocado peel waste and see if the pectinase enzyme produced can be used to make fruit juice clarification.ResultsThe researchers isolated four different bacterial strains from avocado peel waste samples. The potential two bacterial isolates that were identified as being Serratia marcescens and Lysinibacillus macrolides. Finally, the analysis of pectinase production and its application in fruit juice clarification were performed using one of the bacterial strains of Serratia marcescens. The clear apple, lemon, and mango juices were further processed to assess each juice's properties. The highest antioxidant activity was recorded in lemon juice samples. The lemon juice showed the highest total titratable acidity and total phenol content. Apple juices contained the highest total soluble solids, reducing sugar content, and viscosity and the mango juices have the maximum pH value recorded.ConclusionsThe pectinase isolated from the bacterium Serratia marcescens could clear fruit juices. This pectinase needs to be studied more to make sure it works better in the fruit industry and other businesses.

Characterization and optimization of Casimiroa (Casimiroa Edulis) fruit juice using Response Surface Methodology (RSM)

Background: Casimiroa edulis fruit is an underutilized seasonal fruit of Ethiopia, is a rich source of many medicinal and nutritional properties. The fruit is found in different parts of the country, Ethiopia. However, there is no clear research evidences of the fruits on post-harvest handling, preservation, processing, and value addition. Purpose of the study: The present study is aimed at the characterization, optimization of enzyme assisted casimiroa fruit juice clarification process based on selected quality parameters of the final product. Methodology: D-Optimal Mixture Design and RSM were used to investigate level of ingredients and the effects of independent variables on physicochemical composition, functional properties, mineral values and acceptability tests of casimiroa fruit juice. Graphical and numerical optimization were conducted to find the best variables conditions. Casimiroa fruit was treated at various enzyme concentrations (0.01-0.2%), incubation temperature (30-55oC) and incubation time (30-100 mini.) of treatment. A second order Central composite (CCD) was employed to identify the effect of independent variables on the final product. Results: In this study, a statistical significant difference (p < 0.05) was observed within the process variables on nutritional profiles of casimiroa fruit juice. Furthermore, the enzyme treatment reduced the cloudy appearance of casimiroa fruit juice and improved the clarity of juice. The coefficient of determination, R2 values for the dependent variables were almost greater than 0.8. This study revealed that, clarity, viscosity, TSS, TA, pH, phenolic compounds, flavonoids, β-carotene, ascorbic acid, Na, K, Ca, Fe, Zn, P and overall acceptability were significantly (p < 0.05) correlated to incubation temperature, incubation time and enzyme concentration. Incubation time and enzyme concentration were the most important factors affecting the characteristics of the casimiroa fruit juice as it exerted a highly significant influence (p < 0.05) on the dependent variables. Conclusion: In conclusion, enzyme-treatment improves qualities of fruit juice and the overall optimization suggests that enzymatic-treated juice made with 75% fruit pure, 21.25% water, 2.63% sugar and 1.13% citric acid with process variables of incubation temperature (44.01oC), incubation time (62.74 min.) and enzyme concentration (0.2%), respectively achieved the best formulation for this combination of variables with a desirable of 0.9029.

Immobilization, biochemical, thermodynamic, and fruit juice clarification properties of lignocellulosic biomass–derived exo-polygalacturonase from Penicillium paxilli

Immobilization, biochemical, thermodynamic, and fruit juice clarification properties of lignocellulosic biomass–derived exo-polygalacturonase from Penicillium paxilli

Polygalacturonase from Aspergillus japonicus (PGAj): Enzyme production using low-cost carbon source, biochemical properties and application in clarification of fruit juices

Aspergillus japonicus polygalacturonase (PGAj) was produced using passion fruit peel, a residue from the agricultural and juice industries, as a low-cost carbon source. The biochemical properties of the enzyme and its application in the clarification of fruit juices were investigated. The maximum PGAj production was obtained using solid state fermentation (SSF) in 48 h (227.75 ± 13.1 U). PGAj was purified in two chromatography steps with a specific activity of 7.9 U/mg protein, 2.9 -fold purification, and 81% final yield, exhibiting a molecular weight about 40 kDa. The optimal pH and temperature for PGAj activity were pH 4.0 and 55 °C, respectively. Moreover, the PGAj enzyme retained up to 90% of its initial activity for 4 h at pH 4.0, 5.0 and 6.0. The enzyme maintained 83% residual activity after 20 min at 50 °C. PGAj best preferred citrus pectin (8.45 ± 1.6 U/mg protein) as substrate. The best results for juice clarification using 3 U/mL PGAj were obtained with the pulp of Palmer and Tommy mango varieties, white guava, banana, and apple, with increases of 65%, 41%, 40%, 11%, and 9.4% in transmittance values, respectively, compared to the control. This was superior to that obtained using commercial pectinase for some of the pulps, as 49% and 21% for Palmer and Tommy mango pulps, respectively. Thus, PGAj may be suitable for application as a clarifying agent in the fruit juice industry. • Polygalacturonase (PGAj) was produced using a low-cost carbon source (passion fruit peel). • PGAj was obtained in 48 h solid state fermentation (SSF). • Better clarification of mango juice achieved with PGAj than with commercial pectinase.

Microfluidic spinning of fucoxanthin-loaded nanofibers for enhancing antioxidation and clarification of fruit juice.

Fruit juice is one of the most popular drinks, which requires strict processing conditions to ensure its quality, especially to prevent enzymatic browning and turbidity loss. In this work, a new strategy for the preparation of composite nanofibers for juice clarification and anti-browning control was proposed. The strategy used microfluidic spinning to combine Fucoxanthin (Fx), hydroxypropyl-γ-cyclodextrin (HP-γ-CD) and polyvinyl pyrrolidone (PVP) to prepare Fx/HP-γ-CD-PVP (PCF) nanofibers, which not only reflected the excellent antioxidant properties of cyclodextrin-wrapped Fx, but also achieved a more optimized juice clarification agent dosage. Molecular docking technique was used to prove that the stable inclusion complex of Fx and HP-γ-CD could be formed by hydrogen bonding when the molar ratio of Fx to HP-γ-CD was 1 : 2, and the binding energy was as low as -10.23 kcal mol-1. SEM, XRD, FT-IR and TGA were used to characterize the structure of the composite nanofibers, which showed that the thermal stability and water solubility of the embedded Fx were improved. Further studies showed that the apple juice with PCF nanofibers containing inclusion complexes of Fx and HP-γ-CD at a molar ratio of 1 : 2 (PCF 1 : 2) could significantly improve the DPPH and ABTS radical scavenging activity, and could significantly protect the cell membrane integrity of RAW264.7 cells against H2O2 oxidative damage. Finally, the effects of PCF nanofibers on the quality of fresh juice were studied, including clarification experiment and sensory evaluation. The results showed that the dosage of PVP in PCF 1 : 2 was only about 4% of the conventional dosage, and the browning index of fresh juice was significantly reduced with the best clarification. The available data provided in this study would provide a promising safety strategy for the food processing of fresh juice and the extension of its storage life.

EXTRACELLULAR ALKALINE PECTINASES PRODUCTION: A REVIEW

Pectinases are most significant enzymes of commercial sector. Among all the commercially produced industrial enzymes, pectinases occupy 25% share in the overall enzyme market. Pectinases have wide applications in various processes such as fruit juice clarification, wastewater remediation and textile and paper industry etc. Pectinase is most suitable environment friendly tool of nature for sustainable development. Although, pectinases alike many other commercial enzymes are facing the imperative of low harvest and yield. Hence, pectinases have been the objective of research to accomplish the commercial scale articulation levels. To find unique physicochemical properties of pectinases often microbial pectinases are utilized among all the natural sources due to ease in production. Present study primarily centers on the different types of recent techniques for the alkaline pectinases production and optimization strategies advancement for high production yield. For achieving higher production of the extracellular alkaline pectinases by utilizing basic fermentation media and simple molecular devices along with genetic engineering approaches are included in the present study. For the improved alkaline pectinase production advanced genetic engineering techniques such as gene cloning, tools of r-DNA technology and cell immobilization technique are discussed.

Statistical Optimization of Media Components for Xylanase Production by Aspergillus spp. Using Solid State Fermentation and its Application in Fruit Juice Clarification

Xylanases are enzymes that convert xylan into xylose, xylobiose, and xylotriose. The present study deals with the production and optimization of xylanase through Solid-State Fermentation (SSF) using different agricultural wastes by Aspergillus spp. The Plackett Burman (PB) design was used to screen significant media components affecting the xylanase production. The carbon sources screened were wheat bran, rice bran, sugarcane bagasse, corn cob, and orange peel. The nitrogen sources screened were yeast extract, peptone, (NH4)2SO4, Na2NO3, and urea. Also, nine different salts such as KCl, MgSO4, Na2HPO4, CaCl2, FeSO4, ZnSO4, Na2CO3, KH2PO4, and NaH2PO4 which act as trace elements were screened. The results showed that wheat bran, yeast extract, Na2NO3 and KCl are the significant factors that affect xylanase production. A 33 Full Factorial Design (FFD) was performed to optimize the significant media components (wheat bran, KCl, yeast extract) obtained from PB design using Response Surface Methodology (RSM). Statistical analysis of results showed that wheat bran, KCl, yeast extract, and interaction between wheat bran and yeast extract were found to be significant. The optimum concentration of wheat bran, KCl, yeast extract was 8 g/L, 0.1 g/L and 3 g/L. The Partial purification of xylanase was carried out using ammonium salt precipitation and dialysis. Gel filtration chromatography was performed to optimize the elution time, which was found to be 6 minutes. Application of xylanase in orange juice clarification was studied at 40 °C, 50 °C, and 60 °C. The optimum temperature obtained was 60 ºC.

Optimization of Xylanase Production from Aspergillus spp. under Solid-state Fermentation Using Lemon Peel as Substrate

Xylanase is an enzyme that acts on xylan and degrades it into xylose. Xylanase may be produced by several different microbes such as bacteria and fungi. Xylanase has significant applications in food industries for improving dough handling and improving the quality of baked products in the extraction of starch, coffee, and plant oils. Xylanase, along with pectinase and cellulose, has enormous application in the clarification of fruit juices. In the present study, xylanase was produced by solid-state fermentation using lemon peel as substrates. Aspergillus Niger was induced to produce xylanase by frequent sub-culturing on a medium containing 2% xylan. OFAT was analyzed by using several fermentation parameters such as moisture content, particle size, incubation period, incubation temperature, peptone concentration, and pH of the extraction buffer. Under optimized conditions, the maximum xylanase activity was found for particle size of 1.7mm, moisture content of 80%, peptone concentration in nutrient solution at 0.3%, and extraction pH of 7.0. Hence, xylanase can be further subjected to down streaming processes and therefore for various applications.

Kinetic and Thermodynamic Characterization of two Polygalacturonases Isolated from the Digestive Juice of the Snail Limicolaria flammea

Polygalacturonases are extensively used in food industries for pectic substances degradation. In this paper, we investigate on thermal stability parameters of two Polygalacturonases previously isolated from digestive juice of the snail Limicolaria flammea for several industrial applications such as fruit juice clarification. Thermal inactivation was carried out in the temperature range of 55°C to 80°C from 15 to 120 min. All results were statistically analysed. The results shown that thermal inactivation of studied acid phosphatases follows first order kinetics. At their optimum temperatures, these enzymes showed high half-lives ranging from 462.06 to 630.10 min and D values from 1535.00 to 2093.64 min suggesting that these two enzymes had a large thermal stability. The high values of ΔG# (93.96 to 94.97 kJ/mol) reveal a better resistance to denaturation. The relatively high activation energies (from 120.35 to 129.13 kJ/mol) and average enthalpy values (from 117.67 to 126.44 kJ.mol−1) could corroborate the good stability of these biocatalyst. All these results suggest that Polygalacturonases from digestive juice of the snail Limicolaria flammea may be profitably exploited in future food industrial applications.