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At least 883 records · Page 49Linked to original sources
PubMed2000

Analysis of different de-esterification mechanisms for pectin by enzymatic fingerprinting using endopectin lyase and endopolygalacturonase II from A. niger.

A series of pectins with different distribution patterns of methyl ester groups was produced by treatment with either plant (p-PME) or fungal pectin methyl esterases (f-PME) and compared with those obtained by base catalysed de-esterification. The products generated by digestion of these pectins with either endopectin lyase (PL) or endopolygalacturonase II (PG II) from Aspergillus niger were analysed using matrix assisted laser desorption ionisation mass spectrometry (MALDIMS) and high-performance anion-exchange chromatography with pulsed amperometric or UV detection (HPAEC-PAD/UV). Time course analysis using MALDIMS was used to identify the most preferred substrate for each enzyme. For PL, this was shown to be fully methyl esterified HG whereas for PG II, long regions of HG without any methyl esterification, as produced by p-PME was the optimal substrate. The blockwise de-esterification caused by p-PME treatment gave a decrease of partly methylated oligomers in PL fingerprints, which did not effect the relative composition of partly methylated oligomers. PG II fingerprints showed a constant increase of monomers and oligomers without any methyl ester groups with decreasing degree of esterification (DE), but almost no change in the concentration of partly methylated compounds. PL fingerprints of f-PME and chemically treated pectins showed decreasing amounts of partly methyl esterified oligomers with decreasing DE, together with a relative shift towards longer oligomers. PG II fingerprints were characterised by an increase of partly methylated and not methylated oligomers with decreasing DE. But differences were also seen between these two forms of homogenous de-esterification. Introduction of a certain pattern of methyl ester distribution caused by selective removal of certain methyl ester groups by f-PME is the most reasonable explanation for the detected differences.

Aspergillus niger↗
PubMed2001

L-Altruronic acid formed by epimerization of D-galacturonic acid methyl esters during saponification of citrus pectin.

While searching for oligosaccharides containing rhamnose residues in the endopolygalacturonase (EPG) digest of saponified citrus pectin, we found several oligomers containing, in addition to galacturonic acid, a sugar previously unreported in pectin. The 1- and 2-D 1H NMR spectra of the oligosaccharides were consistent with the sugar being a uronic acid with its 2- and 3-hydroxyls being axial and 4-hydroxyl being equatorial. MALDI-TOF mass spectrometry indicated that the oligomers consisted solely of uronic acids. Reduction of the uronic acids in the oligosaccharides converted them to galactose and altrose. The altrose was found to be the L enantiomer by comparison of its trimethylsilyl (-)-2-butyl glycosides to those of authentic D-altrose and a racemic mixture. The sugar was not found in oligosaccharides prepared from EPG digestion of citrus pectin deesterified with pectin methylesterase rather than saponification. Thus, it appears that during saponification, a small proportion of the methylesterified galacturonic acid residues in pectins is epimerized at C-5 leading to formation of L-altruronic acid residues.

Carbohydrate Conformation↗
PubMed2001

Simulation of endo-PG digest patterns and implications for the determination of pectin fine structure.

Novel stepwise approaches to the calculation of enzyme digest patterns are described and used in the validation of a computer simulation. Results obtained using the simulation show that, while a previously proposed model of endo-PG action captures some of the salient features of this enzymes behaviour, it is not sufficient to successfully predict experimental digest patterns from pectic substrates. Subsequently, it has been shown that a modified model incorporating existing information regarding subsite architecture and speculative site tolerances for esterified residues, goes someway towards improving the situation.

Computer Simulation↗
PubMed2003

Low molecular weight chitosans--preparation by depolymerization with Aspergillus niger pectinase, and characterization.

The viscosity of a chitosan solution was rapidly lowered in the presence of pectinase from Aspergillus niger at pH 3.0 and 37 degrees C. The low molecular weight chitosans (LMWC) had a molecular weight in the range 20,000-5000 Da. Circular dichroism spectra showed a decrease in the segment of acetylated glucosamine units, whereas X-ray diffraction and CP-MAS 13C NMR indicated higher crystallinity and polymorphism in LMWC. The latter on thermal drying resulted in structural alterations, and yielded an insoluble product. FT-IR and X-ray diffraction showed no evidence of either Schiff's base linkage or any annealed polymorph. CP-MAS 13C NMR showed marked changes in the chain conformations of LMWC, which are believed to be responsible for its loss of solubility and functionality.

Acetylation↗
PubMed1997

Rhamnogalacturonan II, a dominant polysaccharide in juices produced by enzymic liquefaction of fruits and vegetables.

Rhamnogalacturonan II (RG-II), a small complex pectic polysaccharide, is released from apple (Malus domestica), carrot (Daucus carota), and tomato (Solanum lycopersicum) by treatment with two commercial liquefying enzyme preparations. RG-II was isolated by size-exclusion chromatography from apple, tomato, and carrot juices obtained by enzymic liquefaction. All the RG-IIs contained the diagnostic sugars, apiose, 2-O-methyl-L-fucose, 2-O-methyl-D-xylose, aceric acid, Kdo and Dha. Glycosyl-linkage compositions of the neutral and acidic sugars, including aceric acid, were consistent with the hypothetical model described for sycamore RG-II confirming the conservation of RG-II in plants. Thus, when pectinolytic enzyme preparations are used to process fruits and vegetables, RG-II is released as a main soluble polysaccharide fraction while other pectic polysaccharides are heavily degraded.

Carbohydrate Conformation↗
PubMed2000

Characterization of a novel endopolygalacturonase from Aspergillus niger with unique kinetic properties.

We isolated and characterized a new type of endopolygalacturonase (PG)-encoding gene, pgaD, from Aspergillus niger. The primary structure of PGD differs from that of other A. niger PGs by a 136 amino acid residues long N-terminal extension. Biochemical analysis demonstrated extreme processive behavior of the enzyme on oligomers longer than five galacturonate units. Furthermore, PGD is the only A. niger PG capable of hydrolyzing di-galacturonate. It is tentatively concluded that the enzyme is composed of four subsites. The physiological role of PGD is discussed.

Aspergillus niger↗
PubMed2003

Structural insights into the processivity of endopolygalacturonase I from Aspergillus niger.

Endopolygalacturonase I is a processive enzyme, while the 60% sequence identical endopolygalacturonase II is not. The 1.70 A resolution crystal structure of endopolygalacturonase I reveals a narrowed substrate binding cleft. In addition, Arg96, a residue in this cleft previously shown to be critical for processivity, interacts with the substrate mimics glycerol and sulfate in several well-defined conformations in the six molecules in the asymmetric unit. From this we conclude that both Arg96 and the narrowed substrate binding cleft contribute to retaining the substrate while it moves through the active site after a cleavage event has occurred.

Arginine↗
PubMed2003

Separation of enzymes by sequential macroaffinity ligand-facilitated three-phase partitioning.

Pectinase and cellulase were separated from a commercial enzyme preparation called Pectinex Ultra SP-L. This was carried out using a process called macroaffinity ligand-facilitated three-phase partitioning (MLFTPP). In this method, a water-soluble polymer is floated as an interfacial precipitate by adding ammonium sulfate and tert.-butanol. The polymer (appropriately chosen) in the presence of an enzyme for which it shows affinity, selectively binds to the enzyme and floats as a polymer-enzyme complex. In the first step, pectinase was purified (with alginate as the polymer) 13-fold with 96% activity recovery. In the second MLFTPP step, using chitosan, cellulase was purified 16-fold with 92% activity recovery. Both preparations showed a single band on sodium dodecylsulfate-polyacrylamide gel electrophoresis. This illustrative example shows that the strategy of sequential MLFTPP can be used to separate important biological activities from a crude broth.

Affinity Labels↗
PubMed1997

Fractionation of glycoside precursors of aroma in grapes and wine.

The glycosides in mono-, di- and trihydroxylated terpene and norisoprenoid alcohols and also those in the related shikimate pathway have been isolated on C18 reversed-phase cartridges and then fractionated into classes of different polarity at increasing percentages of methanol. The benzyl alcohol glycosides are the most polar, while those of terpene monohydroxylated alcohols and geranic acid are the least polar. The terpene diols, linalool furanoid and pyranoid oxides and also norisoprenoid precursors show intermediate polarity and separate into well defined fractions according to their polarity.

Butadienes↗
PubMed1976

Pectic enzymes.

Explore the source record for details and available documents.

Bacteria↗
PubMed2001

Enzymatic isolation and structural characterisation of polymeric suberin of cork from Quercus suber L.

An enzymatic method has been used to isolate, for the first time, polymeric suberin from the bark of Quercus suber L. or cork. This was achieved by solvent extraction (dichloromethane, ethanol and water), followed by a step-by-step enzymatic treatment with cellulase, hemicellulase and pectinase, and a final extraction with dioxane/water. The progress of suberin isolation was monitored by Fourier transform infrared spectroscopy using a photoacoustic cell (FTIR-PAS). The material obtained (polymeric suberin (PS)) was characterised by solid-state and liquid-state nuclear magnetic resonance, FTIR-PAS and vapour pressure osmometry, and compared with the suberin fraction obtained by alkaline depolymerisation (depolymerised suberin (DS)). The results showed that PS is an aliphatic polyester of saturated and unsaturated fatty acids, with an average molecular weight (M(w)) of 2050 g mol(-1). Although this fraction represents only 10% of the whole suberin of cork, its polymeric nature gives valuable information about the native form of the polymer. DS was found to have an average M(w) of 750 g mol(-1) and to comprise a significant amount of acidic and alcoholic short aliphatic chains.

Bacterial Proteins↗
PubMed2002

Hydrolysis of pectins with different degrees and patterns of methylation by the endopolygalacturonase of Fusarium moniliforme.

The mode of action of the endopolygalacturonase from Fusarium moniliforme was studied towards a series of pectins with different amounts and distribution patterns of methyl-ester groups. The enzyme hydrolysed the linkages between two galacturonic acid residues according to a multi-chain attack mechanism, at least at the early stage of the reaction. The final percentage of hydrolysis decreased with increasing the degree of methylation. The distribution pattern of the methyl groups affected the rate of hydrolysis as well as the final percentage of hydrolysis, a blockwise distribution being more favourable than a random one. The final products, as analysed by mass spectrometry, included methyl-esterified oligogalacturonates. The detailed analysis of the structure of the oligomers showed that the enzyme was able to accommodate methylated galacturonic acid in its active site, but that methyl-esterification negatively affected the affinity of the enzyme.

Fusarium↗
PubMed2001

Cultivation of Polyporus squamosus for pectinase production in aqueous two-phase system containing sugar beet extraction waste.

Cultivation of the fungus Polyporus squamosus for pectinase production was studied in a polyethylene glycol/crude dextran aqueous two-phase system, with sugar beet extraction waste as pectin source. Fungal growth was restricted to the bottom phase and the amounts of biomass and exo-pectinase activity produced were superior to in homogeneous cultivation. The partition coefficients of endo-pectinase and exo-pectinase were 4.26 and 2.78, respectively. The top phase yields in the single extraction step were about 90% for both pectinases.

Chenopodiaceae↗
PubMed2001

Enzyme-retting of flax and characterization of processed fibers.

Enzyme-retting formulations consisting of Viscozyme L, a pectinase-rich commercial enzyme product, and ethylenediaminetetraacetic acid (EDTA) were tested on Ariane fiber flax and North Dakota seed flax straw residue. Flax stems that were crimped to disrupt the outer layers were soaked with various proportions of Viscozyme-EDTA solutions, retted, and then cleaned and cottonized with commercial processing equipment. Fiber properties were determined and crude test yarns were made of raw and Shirley cleaned flax fibers and cotton in various blend levels. Cleaned fibers were obtained from both seed and fiber flax types, but with variations due to treatment. Retting formulations produced fibers having different properties, with enzyme levels of 0.3% (v/v as supplied) giving finer but weaker fibers than 0.05% regardless of EDTA level. Experimental yarns of blended flax and cotton fibers varied in mass coefficient of variation, single end strength, and nep imperfections due to sample and formulation. With cost and fiber and yarn quality as criteria, results established a range in the amounts of components comprising retting formulations as a basis for further studies to optimize enzyme-retting formulations for flax. Under conditions examined herein, Viscozyme L at 0.3% (v/v) plus 25 mM EDTA produced the best test yarns and, therefore, established a base for future studies to develop commercial-grade, short staple flax fibers for use in textiles.

Edetic Acid↗
PubMed2001

Enzyme processing of textiles in reverse micellar solution.

Scouring of cotton using pectinase enzyme, bioscouring, in reverse micellar system was studied. The effectiveness of bioscouring was evaluated by measuring weight loss of cotton, analyzing pectin and cotton wax remaining and by wetness testing. Pectinase enzyme showed excellent activity even in organic media, and the effectiveness of scouring was equivalent or better than that achieved by conventional alkaline process or bioscouring in aqueous media. Enzymatic modification of wool using protease enzyme in the same system was also studied. It has found that felting property and tensile strength of wool fabrics treated by protease in reverse micellar system were superior to those in aqueous media. Possibilities of utilization of the same system for the subsequent textile dyeing process were also investigated. It was found that cotton and polyester fabrics were dyed satisfactorily by reverse micellar system compared to conventional aqueous system.

Animals↗
PubMed2001

Enzymes and chelating agent in cotton pretreatment.

Desized cotton fabric and cotton seed-coat fragments (impurities) have been treated with commercial cellulase (Celluclast 1.5 L), hemicellulase-pectinase (Viscozyme 120 L) and xylanase (Pulpzyme HC) enzymes. Seed-coat fragments hydrolyzed much faster than the cotton fabric itself. This relative difference in hydrolysis rates makes possible a direct enzymatic removal of seed-coat fragments from desized cotton fabric. Addition of chelating agents such as ethylenediamine-tetra-acetic acid (EDTA) markedly enhanced the directed enzyme action. Pretreatments carried out in acidic solution at pH 5 increased the lightness of seed-coat fragments, contrary to the samples treated in neutral medium at pH 7. Alkaline scouring resulted in darker seed-coat fragments except for the samples pretreated with Pulpzyme HC plus EDTA. This effect is similar to that observed in the biobleaching process in pulp and paper industry.

Calcium↗