D-Glucuronic/D-Galacturonic Acid Assay Kit

Reference code: K-URONIC
SKU: 700004354

100 assays (manual) / 1000 assays (microplate) / 1000 assays (auto-analyser)

Content: 100 assays (manual) / 1000 assays (microplate) / 1000 assays (auto-analyser)
Shipping Temperature: Ambient
Storage Temperature: Short term stability: 2-8oC,
Long term stability: See individual component labels
Stability: > 1 year under recommended storage conditions
Analyte: D-Galacturonic Acid, D-Glucuronic Acid
Assay Format: Spectrophotometer, Microplate, Auto-analyser
Detection Method: Absorbance
Wavelength (nm): 340
Signal Response: Increase
Linear Range: 5 to 150 µg of D-glucuronic acid or D-galacturonic acid per assay
Limit of Detection: ~ 15.5 mg/L
Reaction Time (min): ~ 10 min at 25oC or ~ 5 min at 37oC
Application examples: Hydrolysates of plant material and polysaccharides and other materials.
Method recognition: Novel method

The D-Glucuronic/D-Galacturonic test kit is a simple, reliable and accurate method for the measurement and analysis of D-hexuronic acids (specifically D-glucuronic acid and D-galacturonic acid) in plant extracts, culture media/supernatants and other materials.

Note for Content: The number of manual tests per kit can be doubled if all volumes are halved.  This can be readily accommodated using the MegaQuantTM  Wave Spectrophotometer (D-MQWAVE).

See our complete range of mono/disaccharide test kits.

Scheme-K-URONIC URONIC Megazyme

  • Very cost effective 
  • All reagents stable for > 2 years during use 
  • Only test kit available 
  • Simple format 
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing 
  • Standard included 
  • Suitable for manual, microplate and auto-analyser formats
Certificate of Analysis
Safety Data Sheet
FAQs Assay Protocol Data Calculator Validation Report
Megazyme publication

Measurement of available carbohydrates in cereal and cereal products, dairy products, vegetables, fruit and related food products and animal feeds: First Action 2020.07.

McCleary, B. V. & McLoughlin, C. (2021). Journal of AOAC International, qsab019.

Background: The level of available carbohydrates in our diet is directly linked to two major diseases; obesity and Type II diabetes. Despite this, to date there is no method available to allow direct and accurate measurement of available carbohydrates in human and animal foods. Objective: The aim of this research was to develop a method that would allow simple and accurate measurement of available carbohydrates, defined as non-resistant starch, maltodextrins, maltose, isomaltose, sucrose, lactose, glucose, fructose and galactose. Method: Non-resistant (digestible) starch is hydrolysed to glucose and maltose by pancreatic α-amylase and amyloglucosidase at pH 6.0 with shaking or stirring at 37°C for 4 h. Sucrose, lactose, maltose and isomaltose are completely hydrolyzed by specific enzymes to their constituent monosaccharides, which are then measured using pure enzymes in a single reaction cuvette. Results: A method has been developed that allows the accurate measurement of available carbohydrates in all cereal, vegetable, fruit, food, and feed products, including dairy products. Conclusions: A single-laboratory validation was performed on a wide range of food and feed products. The inter-day repeatability (%RSDr) was <3.58% (w/w) across a range of samples containing 44.1 to 88.9% available carbohydrates. The LOD and LOQ obtained were 0.054% (w/w) and 0.179% (w/w), respectively. The method is all inclusive, specific, robust and simple to use. Highlights: A unique method has been developed for the direct measurement of available carbohydrates, entailing separate measurement of glucose, fructose and galactose; information of value in determining the glycemic index of foods.

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Sequential and enzyme-assisted extraction of algal bioproducts from Ecklonia maxima.

Mabate, B. & Pletschke, B. I. (2024). Enzyme and Microbial Technology, 173, 110364.

Brown algae are gaining recognition as sources of bio-compounds with diverse properties and potential applications in the food, nutraceutical, and pharmaceutical industries. Compounds such as polyphenols, alginates and fucoidan possess multiple bioactivities, including antidiabetic, antioxidant, anticancer, anti-inflammatory, and antibacterial properties. Conventional extraction methods provide low yields, posing challenges for the industrial applications of biocompounds. However, innovations are rapidly emerging to address these challenges, and one such approach is enzyme-assisted extraction. Furthermore, extracting single compounds undervalues algal biomass as valuable compounds may remain in the waste. Therefore, the aim of our study was to develop a framework for the sequential and enzyme-assisted extraction of various bio-compounds using the same biomass in a biorefinery process. The Ecklonia maxima algal biomass was defatted, and polyphenols were extracted using solid-liquid extraction with aqueous ethanol. The remaining residue was treated with an enzyme combination (Cellic® Ctec 2 and Viscozyme L) to liberate carbohydrates into solution, where an alginate and fucoidan fraction were isolated. A second alginate fraction was harvested from the residue. The phenolic fraction yielded about 11% (dry weight of extract/dry weight of seaweed biomass), the alginate fraction 35% and the fucoidan fraction 18%. These were analysed using a variety of biochemical methods. Structural analyses, including FTIR, NMR and TGA, were performed to confirm the integrity of these compounds. This study demonstrated that a sequential extraction method for various algal bioproducts is possible, which can pave the way for a biorefinery approach. Furthermore, our study primarily employed environmentally and eco-friendly extraction technologies promoting an environmentally sustainable industrial approach. This approach enhances the feasibility and flexibility of biorefinery operations, contributing to the development of a circular bio-economy.

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Experimental study and modelling of a packed bed bioreactor for urea removal in wines.

Mazzù, R., Tavilli, E. & Fidaleo, M. (2023). Food and Bioproducts Processing, 140, 230-241.

The study involved the development and modeling of a fixed-bed bioreactor for the removal of urea from wines. The reactor, based on the immobilization of acid urease enzyme, was studied under both stationary and non-stationary conditions. The developed model, including internal and external catalyst particle mass transfer, Michaelis-Menten kinetics, convection and dispersion in the liquid along the reactor axis, was able to produce urea concentration profiles in both the solid and liquid phases under various volumetric flow rates and inlet urea concentrations. The experimental results were in good agreement with the model predictions, the mean relative error between simulated and experimental outlet ammonia concentration ranging from 4.1 % to 16.4 %. Model simulations confirmed that in wines the reaction kinetics is of the pseudo-first order and that internal and external catalyst particle diffusion limitations are negligeable. Simulations of the decrease of urea concentration as a function of space velocity for the reactor under study operating in the continuous mode and for three different wines were obtained confirming that urea removal by immobilized urease in wines is more difficult than in sake. The results obtained form the basis for the designing and scaling up of bioreactors for the treatment of wines.

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Changes in the cocoa shell dietary fiber and phenolic compounds after extrusion determine its functional and physiological properties.

Benítez, V., Rebollo-Hernanz, M., Braojos, C., Cañas, S., Gil-Ramírez, A., Aguilera, Y. & Martín-Cabrejas, M. A. (2023). Current Research in Food Science, 6, 100516.

The influence of different extrusion conditions on the cocoa shell (CS) dietary fiber, phenolic compounds, and antioxidant and functional properties was evaluated. Extrusion produced losses in the CS dietary fiber (3-26%), especially in the insoluble fraction, being more accentuated at higher temperatures (160°C) and lower moisture feed (15-20%). The soluble fiber fraction significantly increased at 135°C because of the solubilization of galactose- and glucose-containing insoluble polysaccharides. The extruded CS treated at 160°C-25% of feed moisture showed the highest increase of total (27%) and free (58%) phenolic compounds, accompanied by an increase of indirect (10%) and direct (77%) antioxidant capacity. However, more promising results relative to the phenolic compounds’ bioaccessibility after in vitro simulated digestion were observed for 135°C-15% of feed moisture extrusion conditions. The CS’ physicochemical and techno-functional properties were affected by extrusion, producing extrudates with higher bulk density, a diminished capacity to hold oil (22-28%) and water (18-65%), and improved swelling properties (14-35%). The extruded CS exhibited increased glucose adsorption capacity (up to 2.1-fold, at 135°C-15% of feed moisture) and α-amylase in vitro inhibitory capacity (29-54%), accompanied by an increase in their glucose diffusion delaying ability (73-91%) and their starch digestion retardation capacity (up to 2.8-fold, at 135°C-15% of feed moisture). Moreover, the extruded CS preserved its cholesterol and bile salts binding capacity and pancreatic lipase inhibitory properties. These findings generated knowledge of the CS valorization through extrusion to produce foods rich in dietary fiber with improved health-promoting properties due to the extrusion-triggered fiber solubilization.

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A Novel polysaccharide DSPP-1 from Durian seed: structure characterization and its neuroprotective effects against Alzheimer's disease in a transgenic Caenorhabditis elegans model.

Xiao, C., Chen, T., Yuan, M., Li, Y., & Wang, F. Europe PMC, (2022), In Press.

Plant polysaccharides have attracted much attention because of their various biological activities. The structure characterization of polysaccharide from durian seed and its neuroprotective effects against Alzheimer's disease in a transgenic Caenorhabditis elegans model were conducted in this study. A water-soluble polysaccharide was obtained using atmospheric pressure plasma treatment, and named DSPP-1. DSPP-1 was composed of rhamnose, galactose and galacturonic acid and its molecular weight was 3.765×105 Da. The study in vitro showed that DPPH radical scavenging activity of DSPP-1 was 79.20% and the inhibitory rate on Aβ1-42 aggregation was 24.65%. In vivo results showed that DSPP-1 could decrease abnormal Aβ 1-42 aggregation to delay the paralysis process of AD-nematodes. Moreover, DSPP-1 significantly improved the antioxidant enzyme activities and reduced lipid peroxidation in AD-nematodes. Taken together, these results indicated that DSPP-1 could be used as a potential natural source for the prevention and treatment of AD.

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The Effect of Dekkera bruxellensis Concentration and Inoculation Time on Biochemical Changes and Cellulose Biosynthesis by Komagataeibacter intermedius.

Devanthi, P. V. P., Pratama, F., Kho, K., Taherzadeh, M. J. & Aslanzadeh, S. (2022). Journal of Fungi, 8(11), 1206.

Bacterial Cellulose (BC) is a biopolymer with numerous applications. The growth of BC-producing bacteria, Komagataeibacter intermedius, could be stimulated by Dekkera bruxellensis, however, the effect on BC yield needs further investigation. This study investigates BC production and biochemical changes in the K. intermedius-D. bruxellensis co-culture system. D. bruxellensis was introduced at various concentrations (103 and 106 CFU/mL) and inoculation times (days 0 and 3). BC yield was ~24% lower when D. bruxellensis was added at 103 CFU/mL compared to K. intermedius alone (0.63 ± 0.11 g/L). The lowest BC yield was observed when 103 CFU/mL yeast was added on day 0, which could be compromised by higher gluconic acid production (10.08 g/L). In contrast, BC yields increased by ~88% when 106 CFU/mL D. bruxellensis was added, regardless of inoculation time. High BC yield might correlate with faster sugar consumption or increased ethanol production when 106 CFU/mL D. bruxellensis was added on day 0. These results suggest that cell concentration and inoculation time have crucial impacts on species interactions in the co-culture system and product yield.

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The effect of black rot on grape berry composition.

Kellner, N., Antal, E., Szabó, A. & Matolcsi, R. (2022). Acta Alimentaria, 51(1), 126-133.

Guignardia bidwellii, indigenous to North America, is a significant pathogen of grapes long known in Hungary, infecting only the growing green parts of the vine (leaves, petioles, shoots, and bunches). In the absence of adequate plant protection and extreme weather conditions such as a predominantly humid, warm year, black rot of grapes can be expected. The pathogen can cause high yield losses due to grape rot and reduce wine quality if the infection is severe. The evolution of certain biogenic amine compounds were investigated under the influence of grape black rot. The results obtained showed that they were present in low concentrations from an oenological point of view. Polyphenol composition was consistent with the literature, blackening affected mainly the concentration of catechin. Black rot fungus does not produce β-glucosidase enzyme. In terms of resveratrol content, black rot has no particular effect. However, like Botrytis cinerea, it produces glycerol and, proportionally, gluconic acid in lower concentrations. It can be concluded that black rot of grapes does not cause health problems when introduced into wine processing.

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A Combination approach in inhibiting Type 2 diabetes-related enzymes using Ecklonia radiata fucoidan and acarbose.

Mabate, B., Daub, C. D., Malgas, S., Edkins, A. L. & Pletschke, B. I. (2021). Pharmaceutics, 13(11), 1979.

Although there are chemotherapeutic efforts in place for Type 2 diabetes mellitus (T2DM), there is a need for novel strategies (including natural products) to manage T2DM. Fucoidan, a sulphated polysaccharide was extracted from Ecklonia radiata. The integrity of the fucoidan was confirmed by structural analysis techniques such as FT-IR, NMR and TGA. In addition, the fucoidan was chemically characterised and tested for cell toxicity. The fucoidan was investigated with regards to its potential to inhibit α-amylase and α-glucosidase. The fucoidan was not cytotoxic and inhibited α-glucosidase (IC50 19 µg/mL) more strongly than the standard commercial drug acarbose (IC50 332 µg/mL). However, the fucoidan lacked potency against α-amylase. On the other hand, acarbose was a more potent inhibitor of α-amylase (IC50 of 109 µg/mL) than α-glucosidase. Due to side effects associated with the use of acarbose, a combination approach using acarbose and fucoidan was investigated. The combination showed synergistic inhibition (>70%) of α-glucosidase compared to when the drugs were used alone. The medicinal implication of this synergism is that a regimen with a reduced acarbose dose may be used, thus minimising side effects to the patient, while achieving the desired therapeutic effect for managing T2DM.

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Inositol metabolism regulates capsule structure and virulence in the human pathogen Cryptococcus neoformans.

Wang, Y., Wear, M., Kohli, G., Vij, R., Giamberardino, C., Shah, A., Toffaletti, D. L., Yu, C. A., Perfect, J. R., Casadevall, A. & Xue, C. (2021). Mbio, 12(6), e02790-21.

The environmental yeast Cryptococcus neoformans is the most common cause of deadly fungal meningitis in primarily immunocompromised populations. A number of factors contribute to cryptococcal pathogenesis. Among them, inositol utilization has been shown to promote C. neoformans development in nature and invasion of central nervous system during dissemination. The mechanisms of the inositol regulation of fungal virulence remain incompletely understood. In this study, we analyzed inositol-induced capsule growth and the contribution of a unique inositol catabolic pathway in fungal development and virulence. We found that genes involved in the inositol catabolic pathway are highly induced by inositol, and they are also highly expressed in the cerebrospinal fluid of patients with meningoencephalitis. This pathway in C. neoformans contains three genes encoding myo-inositol oxygenases that convert myo-inositol into d-glucuronic acid, a substrate of the pentose phosphate cycle and a component of the polysaccharide capsule. Our mutagenesis analysis demonstrates that inositol catabolism is required for C. neoformans virulence and deletion mutants of myo-inositol oxygenases result in altered capsule growth as well as the polysaccharide structure, including O-acetylation. Our study indicates that the ability to utilize the abundant inositol in the brain may contribute to fungal pathogenesis in this neurotropic fungal pathogen.

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Efficient production of nutraceuticals and lactic acid from lignocellulosic biomass by combining organosolv fractionation with enzymatic/fermentative routes.

Karnaouri, A., Asimakopoulou, G., Kalogiannis, K. G., Lappas, A. A. & Topakas, E. (2021). Bioresource Technology, 341, 125846.

The aim of this work was to investigate the use of isobutanol as organic solvent for the efficient delignification and fractionation of beechwood through the OxiOrganosolv process in the absence of any catalyst. The results demonstrate that cellulose-rich solid pulp produced after pretreatment is a source of fermentable sugars that can be easily hydrolyzed and serve as a carbon source in microbial fermentations for the production of omega-3 fatty acids and D-lactic acid. The C5 sugars are recovered in the aqueous liquid fractions and comprise a fraction rich in xylo-oligosaccharides with prebiotic potential. The maximum production of optically pure D-lactic from Lactobacillus delbrueckii sp. bulgaricus reached 51.6 g/L (0.57 g/gbiomass), following a simultaneous saccharification and fermentation strategy. Crypthecodenium cohnii accumulated up to 52.1 wt% lipids with a DHA content of 54.1 %, while up to 43.3 % hemicellulose recovery in form of oligosaccharides was achieved in the liquid fraction.

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Characterization of a novel multidomain CE15-GH8 enzyme encoded by a polysaccharide utilization locus in the human gut bacterium Bacteroides eggerthii.

Kmezik, C., Krska, D., Mazurkewich, S. & Larsbrink, J. (2021). Scientific Reports, 11(1), 1-13.

Bacteroidetes are efficient degraders of complex carbohydrates, much thanks to their use of polysaccharide utilization loci (PULs). An integral part of PULs are highly specialized carbohydrate-active enzymes, sometimes composed of multiple linked domains with discrete functions-multicatalytic enzymes. We present the biochemical characterization of a multicatalytic enzyme from a large PUL encoded by the gut bacterium Bacteroides eggerthii. The enzyme, BeCE15A-Rex8A, has a rare and novel architecture, with an N-terminal carbohydrate esterase family 15 (CE15) domain and a C-terminal glycoside hydrolase family 8 (GH8) domain. The CE15 domain was identified as a glucuronoyl esterase (GE), though with relatively poor activity on GE model substrates, attributed to key amino acid substitutions in the active site compared to previously studied GEs. The GH8 domain was shown to be a reducing-end xylose-releasing exo-oligoxylanase (Rex), based on having activity on xylooligosaccharides but not on longer xylan chains. The full-length BeCE15A-Rex8A enzyme and the Rex domain were capable of boosting the activity of a commercially available GH11 xylanase on corn cob biomass. Our research adds to the understanding of multicatalytic enzyme architectures and showcases the potential of discovering novel and atypical carbohydrate-active enzymes from mining PULs.

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The protective effect of Scenedesmus dimorphus polysaccharide as an antioxidant and antiaging agent on aging rat model induced by D-galactose.

Armaini, A. & Imelda, I. (2021). Journal of Applied Pharmaceutical Science, 11(05), 054-063.

Premature skin aging occurs due to the increased formation of reactive oxygen species (ROS), which causes oxidative stress, DNA damage, and collagen degradation. This study investigates the protective effect of Scenedesmus dimorphus polysaccharides (SDP) as an antioxidant and anti-aging agent on an aging rat model induced by D-galactose (D-gal). This study used 48 male Wistar rats divided into six groups: (1) normal control, (2) polysaccharide control, (3) aging control induced by D-gal 0.25 mg/g bw/days, (4) drug control (vitamin E) treated twice with SDP in an aging rat model, (5) D-gal + SDP (0.2 mg/g bw), and (6) D-gal + SDP (0.8 mg/g bw) treated with oral treatment and observed for periods of 2, 4, and 8 weeks. Giving SDP at a dose of 0.8 mg/g bw can increase superoxide dismutases and catalase activity and reduce malondialdehyde after 8 weeks of observation, which is better than giving vitamin E. The treatment of SDP can stimulate collagen synthesis and reduce advanced glycation end products. Histopathology shows an increase in the area of fibrocollagen proportions and deposition from the matrix on giving SDP, which is better than vitamin E, since SDP can repair skin tissue. Thus, SDP can be useful as an antioxidant and anti-aging agent in an aging Wistar rats model.

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A new, quick, and simple protocol to evaluate microalgae polysaccharide composition.

Decamp, A., Michelo, O., Rabbat, C., Laroche, C., Grizeau, D., Pruvost, J. & Gonçalves, O. (2021). Marine Drugs, 19(2), 101.

In this work, a new methodological approach, relying on the high specificity of enzymes in a complex mixture, was developed to estimate the composition of bioactive polysaccharides produced by microalgae, directly in algal cultures. The objective was to set up a protocol to target oligomers commonly known to be associated with exopolysaccharides’ (EPS) nutraceutical and pharmaceutical activities (i.e., rhamnose, fucose, acidic sugars, etc.) without the constraints classically associated with chromatographic methods, while maintaining a resolution sufficiently high to enable their monitoring in the culture system. Determination of the monosaccharide content required the application of acid hydrolysis (2 M trifluoroacetic acid) followed by NaOH (2 M) neutralization. Quantification was then carried out directly on the fresh hydrolysate using enzyme kits corresponding to the main monosaccharides in a pre-determined composition of the polysaccharides under analysis. Initial results showed that the enzymes were not sensitive to the presence of TFA and NaOH, so the methodology could be carried out on fresh hydrolysate. The limits of quantification of the method were estimated as being in the order of the log of nanograms of monosaccharides per well, thus positioning it among the chromatographic methods in terms of analytical performance. A comparative analysis of the results obtained by the enzymatic method with a reference method (high-performance anion-exchange chromatography) confirmed good recovery rates, thus validating the closeness of the protocol. Finally, analyses of raw culture media were carried out and compared to the results obtained in miliQ water; no differences were observed. The new approach is a quick, functional analysis method allowing routine monitoring of the quality of bioactive polysaccharides in algal cultures grown in photobioreactors.

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Investigation of a thermostable multi-domain xylanase-glucuronoyl esterase enzyme from Caldicellulosiruptor kristjanssonii incorporating multiple carbohydrate-binding modules.

Krska, D. & Larsbrink, J. (2020). Biotechnology for Biofuels, 13, 1-13.

Background: Efficient degradation of lignocellulosic biomass has become a major bottleneck in industrial processes which attempt to use biomass as a carbon source for the production of biofuels and materials. To make the most effective use of the source material, both the hemicellulosic as well as cellulosic parts of the biomass should be targeted, and as such both hemicellulases and cellulases are important enzymes in biorefinery processes. Using thermostable versions of these enzymes can also prove beneficial in biomass degradation, as they can be expected to act faster than mesophilic enzymes and the process can also be improved by lower viscosities at higher temperatures, as well as prevent the introduction of microbial contamination. Results: This study presents the investigation of the thermostable, dual-function xylanase-glucuronoyl esterase enzyme CkXyn10C-GE15A from the hyperthermophilic bacterium Caldicellulosiruptor kristjanssonii. Biochemical characterization of the enzyme was performed, including assays for establishing the melting points for the different protein domains, activity assays for the two catalytic domains, as well as binding assays for the multiple carbohydrate-binding domains present in CkXyn10C-GE15A. Although the enzyme domains are naturally linked together, when added separately to biomass, the expected boosting of the xylanase action was not seen. This lack of intramolecular synergy might suggest, together with previous data, that increased xylose release is not the main beneficial trait given by glucuronoyl esterases. Conclusions: Due to its thermostability, CkXyn10C-GE15A is a promising candidate for industrial processes, with both catalytic domains exhibiting melting temperatures over 70°C. Of particular interest is the glucuronoyl esterase domain, as it represents the first studied thermostable enzyme displaying this activity.

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Tailored and synergistic enzyme-assisted extraction of carotenoid-containing chromoplasts from tomatoes.

Lombardelli, C., Liburdi, K., Benucci, I. & Esti, M. (2020). Food and Bioproducts Processing, 121, 43-53.

Despite significant efforts recently made to improve the recovery yield of food pigments from natural sources, the development of green and sustainable biotechnological approaches is currently under investigation. Within the context of circular economy, food wastes represent a cheap source for the recovery of valuable compounds including food ingredients. In this study, a conservative approach consisting in a tailored enzyme-assisted extraction protocol for the recovery of carotenoid-containing chromoplasts from unsold tomatoes in which lycopene is stable and protected against oxidation, has now been developed. A tailored enzymatic mix based on polygalacturonase, pectin lyase, cellulase and xylanase, has been designed taking into account the polysaccharide composition of the tomato cell wall. The optimal process conditions for enhancing the recovery of carotenoids from tomatoes i.e.; temperature, pH, enzymatic mix of total dosage and processing time, have been investigated. The suitable temperature and pH identified by the RSM analysis, were found to be: 45-55°C at pH 5-5.5. The treatment carried out using the total dosage of 25 U/g for 180 min was the most convenient for maximizing the recovery yield [about 4.30 ± 0.08 (mgLyc/Kgtomato)/U as carotenoid-containing chromoplasts and about 5.43 ± 0.04 (mgLyc/Kgtomato)/U as total carotenoids].

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Safety Information
Symbol : GHS05, GHS08
Signal Word : Danger
Hazard Statements : H315, H318, H360
Precautionary Statements : P201, P202, P264, P280, P302+P352
Safety Data Sheet
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