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endo-Polygalacturonanase M2
(Aspergillus aculeatus)

Product code: E-PGALUSP
€0.00

5,000 Units

Prices exclude VAT

This product has been discontinued

Content: 5,000 Units
Shipping Temperature: Ambient
Storage Temperature: 2-8oC
Formulation: In 3.2 M ammonium sulphate
Physical Form: Suspension
Stability: > 4 years at 4oC
Enzyme Activity: endo-Polygalacturonase
EC Number: 3.2.1.15
CAZy Family: GH28
CAS Number: 9032-75-1
Synonyms: polygalacturonase; (1→4)-alpha-D-galacturonan glycanohydrolase
Source: Aspergillus aculeatus
Molecular Weight: 42,000
Concentration: Supplied at ~ 1,000 U/mL
Expression: Purified from Aspergillus aculeatus
Specificity: Random hydrolysis of α-1,4-D-galactosiduronic linkages in pectate and polygalacturonans.
Specific Activity: ~ 150 U/mg (40oC, pH 5.5 on polygalacturonic acid)
Unit Definition: One Unit of endo-polygalacturonanase activity is defined as the amount of enzyme required to release one µmole of galacturonic acid from polygalacturonic acid (2.5 mg/mL) per min in sodium acetate buffer (100 mM), pH 5.5 at 40oC.
Temperature Optima: 50oC
pH Optima: 5.5
Application examples: Applications in carbohydrate research and in the food industry.

This product has been discontinued (read more).

High purity endo-Polygalacturonanase M2 (Aspergillus aculeatus) for use in research, biochemical enzyme assays and in vitro diagnostic analysis.

A replacement product - endo-Polygalacturonanase from Pectobacterium carotovorum (E-PGALPC) is available. This product exhibits similar biochemical properties and also belongs to GH family GH28. This enzyme is produced recombinantly.

Show all Carbohydrate Active enZYme products on our list.

Documents
Certificate of Analysis
Safety Data Sheet
Data Sheet
Publications
Publication

Production and molecular characterization of tailored citrus pectin-derived compounds.

Humerez-Flores, J. N., Kyomugasho, C., Gutiérrez-Ortiz, A. A., De Bie, M., Panozzo, A., Van Loey, A. M., Moldenaers, P. & Hendrickx, M. E. (2022). Food Chemistry, 367, 130635.

In this study, tailored-made citrus pectin-derived compounds were produced through controlled enzymatic and/or chemical modifications of commercial citrus pectin with different degrees of methylesterification (DM) and similar average molecular weight (MW). In the first treatment, degradation of the citrus pectin (CP) materials by endo-polygalacturonase (EPG) yielded pectins with average Mw’s (between 2 and 60 kDa). Separation and identification of the oligosaccharide fraction present in these samples, revealed the presence of non-methylesterified galacturonic acid oligomers with degree of polymerization (DP) 1-5. In the second treatment, exploiting the combined effect of EPG and pectin lyase, compounds with MW between 2 and 21 kDa, containing methylesterified and non-methylesterified polygalacturonans (DP 1-6), were generated. Finally, CP was sequentially modified by chemical saponification and the action of EPG. A sample of DM 11% and MW 2.7 kDa, containing POS (DP 1-5), was produced. Diverse pectin-derived compounds were successfully generated for further studies exploring their functionality.

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Publication

Red Ginseng Inhibits Tau Aggregation and Promotes Tau Dissociation In Vitro.

Shin, S. J., Park, Y. H., Jeon, S. G., Kim, S., Nam, Y., Oh, S. M., Lee, Y. Y. & Moon, M. (2020). Oxidative Medicine and Cellular Longevity, 2020, 7829842.

Tau, a microtubule-associated protein expressed in mature neurons, interacts with tubulin to promote the assembly and stabilization of microtubules. However, abnormally hyperphosphorylated tau dissociates from microtubules and self-aggregates. Tau aggregates, including paired helical filaments and neurofibrillary tangles, promote neuronal dysfunction and death and are the defining neuropathological feature of tauopathies. Therefore, suppressing tau aggregation or stimulating the dissociation of tau aggregates has been proposed as an effective strategy for treating neurodegenerative diseases associated with tau pathology such as Alzheimer’s disease (AD) and frontotemporal dementia. Interestingly, ginsenosides extracted from Panax ginseng reduced the hippocampal and cortical expression of phosphorylated tau in a rat model of AD. However, no studies have been conducted into the effect of red ginseng (RG) and its components on tau pathology. Here, we evaluated the effect of Korean red ginseng extract (KRGE) and its components on the aggregation and disassociation of tau. Using the thioflavin T assay, we monitored the change in fluorescence produced by the aggregation or disassociation of tau K18, an aggregation-prone fragment of tau441 containing the microtubule-binding domain. Our analysis revealed that KRGE not only inhibited tau aggregation but also promoted the dissociation of tau aggregates. In addition, the KRGE fractions, such as saponin, nonsaponin, and nonsaponin fraction with rich polysaccharide, also inhibited tau aggregation and promoted the dissociation of tau aggregates. Our observations suggest that RG could be a potential therapeutic agent for the treatment of neurodegenerative diseases associated with tauopathy.

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Publication

Characterization of a Novel Thermophilic Endopolygalacturonase Produced by Bacillus licheniformis IEB-8.

Hadri, S. H., Asad, M. J., Hyder, M. Z., Naqvi, S. M. S., Mukhtar, T., Zafar, M., Shah, S. H., Mehmood, R. T. & Wu, J. D. (2019). BioResources, 14(2), 2873-2884.

Endopolygalacturonases characterized until now have either low working temperatures, working pH in acidic range, high Michaelis-Menten constant (Km), or a high production cost. These characteristics are a hurdle in the industrial applications of these endopolygalacturonases. The purpose of this work was to characterize a novel endopolygalacturonase produced by Bacillus licheniformis IEB-8. Phylogenetic analysis of Bacillus licheniformis IEB-8 showed that the isolate was unique. Citrus peels were used as the only nutrient source for the growth of Bacillus licheniformis IEB-8, allowing a cheap production of endopolygalacturonase. All the synthetic carbon sources showed a negative impact on the production of endopolygalacturonase, while ammonium sulfate enhanced its production. Among different metal ions, Zn+2 showed a negative effect while Mg+2 and Ca+2 did not have any significant effect on the endopolygalacturonase activity. A Lineweaver-Burk plot was prepared for the characterization of the kinetic parameters including Km and Vmax, which were 0.45 mg/mL and 285.7 µM/min, respectively. A comprehensive comparison of the endopolygalacturonase from this study with the available literature indicated that it is better than the reported and commercially available endopolygalacturonases in having the optimum working temperature of 55°C, a low Km of 0.57 mg/mL, and pH of 7 to 8, which indicated its novelty.

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Publication
Active proton efflux, nutrient retention and boron-bridging of pectin are related to greater tolerance of proton toxicity in the roots of two Erica species.

Oliva, S. R., Mingorance, M. D., Sanhueza, D., Fry, S. C. & Leidi, E. O. (2018). Plant Physiology and Biochemistry, In Press.

Background and aims: Tolerance to soil acidity was studied in two species of Ericaceae that grow in mine-contaminated soils (S Portugal, SW Spain) to find out if there are interspecific variations in H+ tolerance which might be related to their particular location. Methods: Tolerance to H+ toxicity was tested in nutrient solutions using seeds collected in SW Spain. Plant growth and nutrient contents in leaves, stems and roots were determined. Viability tests and proton exchange were studied in roots exposed, short-term, to acidic conditions. Membrane ATPase activity and the cell-wall pectic polysaccharide domain rhamnogalacturonan-II (RG-II) were analysed to find out interspecific differences. Results: Variation in survival, growth and mineral composition was found between species. The H+-tolerant species (Erica andevalensis) showed greater concentration of nutrients than E. australis. Very low pH (pH 2) produced a significant loss of root nutrients (K, P, Mg) in the sensitive species. Root ATPase activity was slightly higher in the tolerant species with a correspondingly greater H+ efflux capacity. In both species, the great majority of the RG-II domains were in their boron-bridged dimeric form. However, shifting to a medium of pH 2 caused some of the boron bridges to break in the sensitive species. Conclusions: Variation in elements linked to the cell wall-membrane complex and the stability of their components (RG-II, H+-ATPases) are crucial for acid stress tolerance. Thus, by maintaining root cell structure, active proton efflux avoided toxic H+ build-up in the cytoplasm and supported greater nutrient acquisition in H+-tolerant species.

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Publication
Pectin oligosaccharides from sugar beet pulp: molecular characterization and potential prebiotic activity.

Prandi, B., Baldassarre, S., Babbar, N., Bancalari, E., Vandezande, P., Hermans, D., Bruggeman, G., Gatti, M., Elst, K. & Stefano Sforza, S. (2018). Food & Function, In Press.

Pectin oligosaccharides (POS) have been indicated as a new class of potential prebiotic compounds, which can be produced from pectin-rich food byproducts. In the present study, different technological means of POS production were explored to produce tailor-made POS mixtures starting from sugar beet pulp. The overall POS production process consisted of two steps: the extraction of pectin and the hydrolysis of pectin to tailored POS by combined hydrolysis/fractionation approaches. Different extraction as well as hydrolysis and fractionation methodologies were applied. The obtained POS were characterized for their total galacturonic acid content and, at a deeper level, using a HILIC-ESI/MS methodology, for the POS structure and composition. The composition of POS fractions was studied as a function of the technology used to obtain them. Finally, the potential prebiotic properties of the POS mixtures obtained were thoroughly explored by several in vitro experiments aimed at detecting lactic acid bacteria (LAB) stimulation by POS fractions. Several fractions were very efficient in stimulation, in a species-dependent manner. The overall best fractions were in general those rich in arabinans having a low degree of polymerization, obtained from the enzymatic extraction of biomass and subsequent fractionation with low-medium molecular weight cut-off. Quite interestingly, no POS fraction was able to stimulate pathogenic E. coli strains. The data reported here clearly indicate the possibility to obtain diverse fractions with different prebiotic properties starting from the same biomass, and outline clear potential for POS obtained from sugar beet pulp with the appropriate technology to act as prebiotic compounds.

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Publication
Active pectin fragments of high in vitro antiproliferation activities toward human colon adenocarcinoma cells: Rhamnogalacturonan II.

Ai, L., Chung, Y. C., Lin, S. Y., Lee, K. C., Lai, P. F. H., Xia, Y., Wang, G. & Cui, S. W. (2018). Food Hydrocolloids, 83, 239-245.

This study was to identify active pectin fragments of high in vitro anti-proliferation activities against human colon adenocarcinoma Caco-2 cells, using various pectin fragment preparations dominant with rhamnogalacturonan I (RG-I), rhamnogalacturonan II (RG-II), and galacturonic acid oligomers (GAOS). The samples were prepared from citrus high and low methoxy pectins (HPE and LPE) by endo-α-D-(1,4)-polygalacturonase action coupling with a green process as ultrafiltration (membrane molecular weight cut off  > 10k, 1k-10k, and < 1k, giving fragment preparations I, II, and III, respectively). It is indicated that, in the fragment preparations HPE-I, II, and III, the molecular fractions HPE-F1 (RG-I), HPE-F2 (RG-II), and HPE-F3(GAOS) showed an averaged Mw = 26.7k, 3.4k, and 0.3k, respectively, notably higher than their LPE counterparts. All fragment preparations exhibited concentration-dependent, inhibition effects on Caco-2 cells, where HPE-II (RG-II-domain) revealed an interesting inhibition effect, significantly higher than those did HPE-I (RG-I domain) and HPE-III (GAOS). The HPE-F2 (RG-II) was elucidated and found of IR = 88% at 1.0 mg/mL and half-inhibition concentration (IC50) = 0.12 mg/mL, doubling the effect of HPE-F1 (RG-I) (IR = 45% at 1.0 mg/mL). Conclusively, citrus RG-II fragment showed a promisingly high anti-Caco-2 activity, significantly higher than did RG-I and GAOS, possibly attributed to its special branched structure and low molecular size.

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Publication
Remodeling of pectin and hemicelluloses in tomato pericarp during fruit growth.

Guillon, F., Moïse, A., Quemener, B., Bouchet, B., Devaux, M. F., Alvarado, C. & Lahaye, M. (2017). Plant Science, 257, 48-62.

Tomato fruit texture depends on histology and cell wall architecture, both under genetic and developmental controls. If ripening related cell wall modifications have been well documented with regard to softening, little is known about cell wall construction during early fruit development. Identification of key events and their kinetics with regard to tissue architecture and cell wall development can provide new insights on early phases of texture elaboration. In this study, changes in pectin and hemicellulose chemical characteristics and location were investigated in the pericarp tissue of tomato (Solanum lycopersicon var Levovil) at four stages of development (7, 14 and 21 day after anthesis (DPA) and mature green stages). Analysis of cell wall composition and polysaccharide structure revealed that both are continuously modified during fruit development. At early stages, the relative high rhamnose content in cell walls indicates a high synthesis of rhamnogalacturonan I next to homogalacturonan. Fine tuning of rhamnogalacturonan I side chains appears to occur from the cell expansion phase until prior to the mature green stage. Cell wall polysaccharide remodelling also concerns xyloglucans and (galacto)glucomannans, the major hemicelluloses in tomato cell walls. In situ localization of cell wall polysaccharides in pericarp tissue revealed non-ramified RG-I rich pectin and XyG at cellular junctions and in the middle lamella of young fruit. Blocks of non-methyl esterified homogalacturonan are detected as soon as 14 DPA in the mesocarp and remained restricted to cell corner and middle lamella whatever the stages. These results point to new questions about the role of pectin RGI and XyG in cell adhesion and its maintenance during cell expansion.

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Publication

Partial purification of a polygalacturonase from a new Aspergillus sojae mutant and its application in grape mash maceration.

Yıldız, S., Mata‐Gómez, M. A., Tarı, C. & Rito‐Palomares, M. (2017). International Journal of Food Science & Technology, 52(3), 834-842.

The use of polygalacturonase (PG) preparations in winemaking promotes the release of phenolic compounds. A PG from a new source, Aspergillus sojae mutant, was semi-purified and tested for grape mash maceration. Crude extract (CE), a commercial pectinase, and two high PG activity semi-purified preparations, FI and FII, were applied for maceration at PG activity of 3.5 U g-1 of grape for 46 h. Enzyme-assisted maceration significantly (P < 0.05) increased the total phenolic content from 255.8 to 916.3 ± 5.2, 5732.9 ± 9.9, 563.4 ± 6.7 and 620.6 ± 18.4 mg L-1 for CE, commercial pectinase, FI and FII, respectively. The content of individual phenolics such as gallic, protocatechuic, chlorogenic and p-coumaric acids was improved. Principal component and hierarchical clustering analyses suggested that CE has a better performance upon the release of phenols. Semi-purified preparations acted similar to commercial pectinase. These findings open an opportunity for the potential use of PG from the mutant strain as an alternative macerating enzyme.

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Publication
Introduction and characterization of charged functional domains into an esterified pectic homogalacturonan by a citrus pectin methylesterase and comparison of its modes of action to other pectin methylesterase isozymes.

Kim, Y., Williams, M. A., Luzio, G. A. & Cameron, R. G. (2017). Food Hydrocolloids, 69, 422-431.

One of the four pectin methylesterase types isolated from Citrus sinensis var. Valencia fruit was used to demethylesterify a model homogalacturonan (HG) to 30%, 50% and 70% degree of methylesterification (DM) at pH 4.5 and 7.0, respectively. Introduced demethylesterified blocks (DMBs) were released by a limited  endo-polygalacturonase (EPG) digestion, separated and quantified by HPAEC. Average DMB size (BS) and number of such blocks per molecule (BN) differed depending on final DM and reaction pH (P < 0.05). BS and BN were significantly higher in 30% DM HG than 50 and 70 DMs. pH 4.5 series showed significantly larger BS compared to pH 7.5 series (P < 0.01). Distribution of DMBs released by limited EPG digest was predicted by mathematical modeling and in silico modeled processive (degree of processivity = 10), multiple attack mode of action best explains the experimental block distributions. Absolute degree of blockiness (DBabs) obtained from exhaustive   EPG digestions, displayed a linear relationship with DM regardless of reaction pH (P < 0.001). Significant correlation coefficients between BS, BN, DBabs, and DM manifested the effectiveness of the block information gained from both EPG digestion to estimate DMB distribution pattern (P < 0.05). However, comparison of block distribution information of three isozymes revealed that difference in block pattern could be manifested by parameters from limited EPG digest (BS, BN ) but not by those from exhaustive digest (DBabs). The results suggested the possibility to control BS and to customize specific population of demethylesterified pectin molecules using PME isozymes from Valencia orange.

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Publication
Cell separation in kiwifruit without development of a specialised detachment zone.

Prakash, R., Hallett, I. C., Wong, S. F., Johnston, S. L., O’Donoghue, E. M., McAtee, P. A., Seal, A. G., Atkinson, R. G. & Schröder, R. (2017). BMC Plant Biology, 17(1), 86.

Background: Unlike in abscission or dehiscence, fruit of kiwifruit Actinidia eriantha develop the ability for peel detachment when they are ripe and soft in the absence of a morphologically identifiable abscission zone. Two closely-related genotypes with contrasting detachment behaviour have been identified. The ‘good-peeling’ genotype has detachment with clean debonding of cells, and a peel tissue that does not tear. The ‘poor-peeling’ genotype has poor detachability, with cells that rupture upon debonding, and peel tissue that fragments easily. Results: Structural studies indicated that peel detachability in both genotypes occurred in the outer pericarp beneath the hypodermis. Immunolabelling showed differences in methylesterification of pectin, where the interface of labelling coincided with the location of detachment in the good-peeling genotype, whereas in the poor-peeling genotype, no such interface existed. This zone of difference in methylesterification was enhanced by differential cell wall changes between the peel and outer pericarp tissue. Although both genotypes expressed two polygalacturonase genes, no enzyme activity was detected in the good-peeling genotype, suggesting limited pectin breakdown, keeping cell walls strong without tearing or fragmentation of the peel and flesh upon detachment. Differences in location and amounts of wall-stiffening galactan in the peel of the good-peeling genotype possibly contributed to this phenotype. Hemicellulose-acting transglycosylases were more active in the good-peeling genotype, suggesting an influence on peel flexibility by remodelling their substrates during development of detachability. High xyloglucanase activity in the peel of the good-peeling genotype may contribute by having a strengthening effect on the cellulose-xyloglucan network. Conclusions: In fruit of A. eriantha, peel detachability is due to the establishment of a zone of discontinuity created by differential cell wall changes in peel and outer pericarp tissues that lead to changes in mechanical properties of the peel. During ripening, the peel becomes flexible and the cells continue to adhere strongly to each other, preventing breakage, whereas the underlying outer pericarp loses cell wall strength as softening proceeds. Together these results reveal a novel and interesting mechanism for enabling cell separation.

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Safety Data Sheet
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