Identification and Characterization of Citrus Peel Uronic Acid Oxidase.

Uronic acid-rich plant materials such as pectin are potential renewable feedstocks for the chemical industry. Uronic acid oxidase activity was first reported in extracts of citrus leaves, and was subsequently found to be widely distributed in plants, with the highest activity detected in extracts of the pectin-rich citrus peel. Herein we report the identification of the primary sequence of uronic acid oxidase from extracts of the peel of Citrus sinensis, by partial purification and protein mass spectrometry. Activity of the enzyme, a member of the berberine bridge family, was confirmed by recombinant expression in Pichia pastoris. Biochemical characterization of the recombinant enzyme is reported. Our findings facilitate further investigation of the biological function of uronic acid oxidation in the economically important orange fruit, and also provide a basis for the development of a catalyst for bioconversion of uronic acids.

Production and Biotechnological Potential of Extracellular Polymeric Substances from Sponge-Associated Antarctic Bacteria.

Four sponge-associated Antarctic bacteria (i.e., Winogradskyella sp. strains CAL384 and CAL396, Colwellia sp. strain GW185, and Shewanella sp. strain CAL606) were selected for the highly mucous appearance of their colonies on agar plates. The production of extracellular polymeric substances (EPSs) was enhanced by a step-by-step approach, varying the carbon source, substrate and NaCl concentrations, temperature, and pH. The EPSs produced under optimal conditions were chemically characterized, resulting in a moderate carbohydrate content (range, 15 to 28%) and the presence of proteins (range, 3 to 24%) and uronic acids (range, 3.2 to 11.9%). Chemical hydrolysis of the carbohydrate portion revealed galactose, glucose, galactosamine, and mannose as the principal constituents. The potential biotechnological applications of the EPSs were also investigated. The high protein content in the EPSs from Winogradskyella sp. CAL384 was probably responsible for the excellent emulsifying activity toward tested hydrocarbons, with a stable emulsification index (E24) higher than those recorded for synthetic surfactants. All the EPSs tested in this work improved the freeze-thaw survival ratio of the isolates, suggesting that they may be exploited as cryoprotection agents. The addition of a sugar in the culture medium, by stimulating EPS production, also allowed isolates to grow in the presence of higher concentrations of mercury and cadmium. This finding was probably dependent on the presence of uronic acids and sulfate groups, which can act as ligands for cations, in the extracted EPSs.IMPORTANCE To date, biological matrices have never been employed for the investigation of EPS production by Antarctic psychrotolerant marine bacteria. The biotechnological potential of extracellular polymeric substances produced by Antarctic bacteria is very broad and comprises many advantages, due to their biodegradability, high selectivity, and specific action compared to synthetic molecules. Here, several interesting EPS properties have been highlighted, such as emulsifying activity, cryoprotection, biofilm formation, and heavy metal chelation, suggesting their potential applications in cosmetic, environmental, and food biotechnological fields as valid alternatives to the commercial polymers currently used.

Quantification of anti-nutritional factors and their correlations with protein and oil in soybeans.

Soybeans contain about 30% carbohydrate, mainly consisting of non-starch polysaccharides (NSP) and oligosaccharides. NSP are not hydrolyzed in the gastrointestinal tract of monogastric animals. These NSP negatively affect the development of these animals, especially the soluble fraction. This work aimed to establish a method to quantify NSP in soybeans, using high performance liquid chromatography (HPLC), and to estimate correlations between NSP, oligosaccharides, protein and oil. Sucrose, raffinose + stachyose, soluble and insoluble NSP contents were determined by HPLC. Oil and protein contents were determined by near-infrared spectroscopy (NIRS). The soluble PNAs content showed no significant correlation with protein, oil, sucrose and raffinose + stachyose contents, but oligosaccharides showed a negative correlation with protein content. These findings open up the possibility of developing cultivars with low soluble NSP content, aiming to develop feed for monogastric animals.

Characterization and Prebiotic Potential of Longan Juice Obtained by Enzymatic Conversion of Constituent Sucrose into Fructo-Oligosaccharides.

The prebiotic potential of longan juice obtained by a commercial Viscozyme L for conversion of constituent sucrose to fructo-oligosaccharide was investigated. The physicochemical properties and carbohydrate composition of the longan juice was evaluated before and after enzymatic treatment. The stimulation effects of the treated longan juice on probiotic bacteria growth were also studied in vitro. The results showed that total soluble solids, yield and clarity of longan juice were all significantly improved after enzyme treatment. The water-soluble polysaccharide content, including pectin, was significantly increased. Compared with the natural longan pulp, the enzyme treated juice showed a significant decrease in sucrose content. Substantial fructo-oligosaccharides including 1-kestose and nystose were synthesized after enzyme treatment. The molecular weight distribution and the monosaccharide composition of the water-soluble polysaccharide were significantly changed by enzyme treatment. The treated longan juice and its ethanol-soluble sugar fraction promoted the growth of Streptococus thermophiles, Lactobacillus acidophilus and Lactobacillus delbrueckii, showing a good potential of the treated longan juice for producing functional foods and nutraceuticals.

Differentiating chondroitin sulfate glycosaminoglycans using collision-induced dissociation; uronic acid cross-ring diagnostic fragments in a single stage of tandem mass spectrometry.

The stereochemistry of the hexuronic acid residues of the structure of glycosaminoglycans (GAGs) is a key feature that affects their interactions with proteins and other biological functions. Electron based tandem mass spectrometry methods, in particular electron detachment dissociation (EDD), have been able to distinguish glucuronic acid (GlcA) from iduronic acid (IdoA) residues in some heparan sulfate tetrasaccharides by producing epimer-specific fragments. Similarly, the relative abundance of glycosidic fragment ions produced by collision-induced dissociation (CID) or EDD has been shown to correlate with the type of hexuronic acid present in chondroitin sulfate GAGs. The present work examines the effect of charge state and degree of sodium cationization on the CID fragmentation products that can be used to distinguish GlcA and IdoA containing chondroitin sulfate A and dermatan sulfate chains. The cross-ring fragments (2,4)A(n) and (0,2)X(n) formed within the hexuronic acid residues are highly preferential for chains containing GlcA, distinguishing it from IdoA. The diagnostic capability of the fragments requires the selection of a molecular ion and fragment ions with specific ionization characteristics, namely charge state and number of ionizable protons. The ions with the appropriate characteristics display diagnostic properties for all the chondroitin sulfate and dermatan sulfate chains (degree of polymerization of 4-10) studied.

Comparative study of two table grape varieties with contrasting texture during cold storage.

Postharvest softening of grape berries is one of the main problems affecting grape quality during export. Cell wall disassembly, especially of pectin polysaccharides, has been commonly related to fruit softening, but its influence has been poorly studied in grapes during postharvest life. In order to better understand this process, the Thompson seedless (TS) variety, which has significantly decreased berry texture after prolonged cold storage, was compared to NN107, a new table grape variety with higher berry firmness. Biochemical analysis revealed a greater amount of calcium in the cell wall of the NN107 variety and less reduction of uronic acids than TS during cold storage. In addition, the activity of polygalacturonase was higher in TS than NN107 berries; meanwhile pectin methylesterase activity was similar in both varieties. Polysaccharide analysis using carbohydrate gel electrophoresis (PACE) suggests a differential pectin metabolism during prolonged cold storage. Results revealed lower pectin fragments in TS after 60 days of cold storage and shelf life (SL) compared to 30 days of cold storage and 30 + SL, while NN107 maintained the same fragment profile across all time points evaluated. Our results suggest that these important differences in cell wall metabolism during cold storage could be related to the differential berry firmness observed between these contrasting table grape varieties.

Culturable diversity and biochemical features of thraustochytrids from coastal waters of Southern China.

Thraustochytrids are ubiquitous marine osmo-heterotrophic fungi-like microorganisms with only about 40 identified species till now. In this study, a total of 60 thraustochytrid strains were isolated from marine coastal habitats. Analysis of 18S rRNA gene sequences revealed that they belonged to three genera, i.e., Schizochytrium, Aurantiochytrium, and Thraustochytrium. All of the isolates were found to show considerable cellulolytic and lipolytic activities. Strains of Aurantiochytrium sp. and Thraustochytrium sp. were found to produce the highest levels of extracellular polysaccharides (EPS), which reached 345 µg ml(-1) in the growth media. Fourier transform infrared (FTIR) spectra of the EPS samples derived from two thraustochytrids (PKU#Sed1 and #SW1) displayed peaks for carbohydrates, proteins, lipids, uronic acids, and nucleic acids. Fatty acid profiles of four thraustochytrids comprised of palmitic acid (C16:0) and docosahexaenoic acid (DHA) as their major constituents. Schizochytrium sp. demonstrated the highest DHA production at 44 % of total fatty acids (TFA) with biomass and DHA yield of 7.1 and 1.6 g l(-1), respectively, on the fourth day of growth. All the four isolates exhibited considerable production of palmitic acid (16:0) in their fatty acid profiles ranging from 35 to 50 % TFA. This is the first report on extracellular enzymes, EPS, and DHA production from thraustochytrids isolated from the coastal habitats of China.

Differences between easy- and difficult-to-mill chickpea (Cicer arietinum L.) genotypes. Part III: free sugar and non-starch polysaccharide composition.

BACKGROUND: Parts I and II of this series of papers identified several associations between the ease of milling and the chemical compositions of different chickpea seed fractions. Non-starch polysaccharides were implicated; hence, this study examines the free sugars and sugar residues. RESULTS: Difficult milling is associated with: (1) lower glucose and xylose residues (less cellulose and xyloglucans) and more arabinose, rhamnose and uronic acid in the seed coat, suggesting a more flexible seed coat that resists cracking and decortication; (2) a higher content of soluble and insoluble non-starch polysaccharide fractions in the cotyledon periphery, supporting a pectic polysaccharide mechanism comprising arabinogalacturonan, homogalacturonan, rhamnogalalcturonan, and glucuronan backbone structures; (3) higher glucose and mannose residues in the cotyledon periphery, supporting a lectin-mediated mechanism of adhesion; and (4) higher arabinose and glucose residues in the cotyledon periphery, supporting a mechanism involving arabinogalactan-proteins. CONCLUSION: This series has shown that the chemical composition of chickpea does vary in ways that are consistent with physical explanations of how seed structure and properties relate to milling behaviour. Seed coat strength and flexibility, pectic polysaccharide binding, lectins and arabinogalactan-proteins have been implicated. Increased understanding in these mechanisms will allow breeding programmes to optimise milling performance in new cultivars.

Differences between easy- and difficult-to-mill chickpea (Cicer arietinum L.) genotypes. Part I: broad chemical composition.

BACKGROUND: Ease of milling is an important quality trait for chickpeas (Cicer arietinum L.) and involves two separate processes: removal of the seed coat and splitting of cotyledons. Four chickpea genotypes (two desi types, one kabuli type and one interspecific hybrid with 'wild' C. echinospermum parentage) of differing ease of milling were examined to identify associated seed composition differences in the seed coat, cotyledons and their junctions (abaxial and adaxial). RESULTS: Several components in different fractions were associated with ease of milling chickpea seeds: primarily soluble and insoluble non-starch polysaccharides (including pectins) and protein at the seed coat and cotyledon junctions, and the lignin content of the seed coat. CONCLUSION: This study shows that the chemical composition of chickpea does vary with seed type (desi and kabuli) and within desi genotypes in ways that are consistent with physical explanations of how seed structure and properties relate to milling behaviour.

Chondroitin sulphate distribution in broiler chicken carcasses.

1. The content of chondroitin sulphate (CS), known as a nutraceutical, was estimated in broiler chicken carcasses by analysing sulphated glycosaminoglycan uronic acid in posterior sternum (keel) cartilage and bones from 4 parts (wing, leg, front and hind) of carcasses. 2. The results of the present study suggested that approximately 0.63 g CS uronic acid (or 1.9 g as CS) can be extracted from a 1.66 kg whole broiler chicken carcass. The amount of extractable CS from keel cartilage, which has been reported as a valuable source of CS in broiler chicken carcasses, was surprisingly low (<10% of total CS).

Comparison of kink-band structures and specificities of cell wall polysaccharides in modern and ancient flax fibres.

Flax (Linum usitatissimum L.) is a plant of industrial importance, its fibres being presently used for high-value textile applications, composite reinforcements as well as natural actuators. Human interest in this fibre-rich plant dates back several millennia, including to Ancient Egypt where flax was used extensively in various quotidian items. While the recent technical developments of flax fibres continue to diversify through scientific research, the historical use of flax also has rich lessons for today. Through careful examination of ancient Egyptian and modern flax fibres, this study aims to conduct a multi-scale characterization from the yarn to the fibre cell wall scale, linking differences in structure and polysaccharide content to the mechanical performance and durability of flax. Here, a multi-scale biochemical study is enriched by scanning electron microscopy and nanomechanical investigations. A key finding is the similarity of cellulose features, crystallinity index and local mechanical performances between ancient and modern fibres. Biochemically speaking, monosaccharides analysis, deep-UV and NMR investigations demonstrate that ancient fibres exhibit less pectins but a similar hemicellulosic content, especially through uronic acids and galactose, suggesting the sensitivity of these non-crystalline components.

Conditional requirement for exopolysaccharide in the Mesorhizobium-Lotus symbiosis.

Rhizobial surface polysaccharides are required for nodule formation on the roots of at least some legumes but the mechanisms by which they act are yet to be determined. As a first step to investigate the function of exopolysaccharide (EPS) in the formation of determinate nodules, we isolated Mesorhizobium loti mutants affected in various steps of EPS biosynthesis and characterized their symbiotic phenotypes on two Lotus spp. The wild-type M. loti R7A produced both high molecular weight EPS and lower molecular weight (LMW) polysaccharide fractions whereas most mutant strains produced only LMW fractions. Mutants affected in predicted early biosynthetic steps (e.g., exoB) formed nitrogen-fixing nodules on Lotus corniculatus and L. japonicus 'Gifu', whereas mutants affected in mid or late biosynthetic steps (e.g., exoU) induced uninfected nodule primordia and, occasionally, a few infected nodules following a lengthy delay. These mutants were disrupted at the stage of infection thread (IT) development. Symbiotically defective EPS and Nod factor mutants functionally complemented each other in co-inoculation experiments. The majority of full-length IT observed harbored only the EPS mutant strain and did not show bacterial release, whereas the nitrogen-fixing nodules contained both mutants. Examination of the symbiotic proficiency of the exoU mutant on various L. japonicus ecotypes revealed that both host and environmental factors were linked to the requirement for EPS. These results reveal a complex function for M. loti EPS in determinate nodule formation and suggest that EPS plays a signaling role at the stages of both IT initiation and bacterial release.

Fibres from flax overproducing ß-1,3-glucanase show increased accumulation of pectin and phenolics and thus higher antioxidant capacity.

BACKGROUND: Recently, in order to improve the resistance of flax plants to pathogen infection, transgenic flax that overproduces ß-1,3-glucanase was created. ß-1,3-glucanase is a PR protein that hydrolyses the ß-glucans, which are a major component of the cell wall in many groups of fungi. For this study, we used fourth-generation field-cultivated plants of the Fusarium -resistant transgenic line B14 to evaluate how overexpression of the ß-1,3-glucanase gene influences the quantity, quality and composition of flax fibres, which are the main product obtained from flax straw. RESULTS: Overproduction of ß-1,3-glucanase did not affect the quantity of the fibre obtained from the flax straw and did not significantly alter the essential mechanical characteristics of the retted fibres. However, changes in the contents of the major components of the cell wall (cellulose, hemicellulose, pectin and lignin) were revealed. Overexpression of the ß-1,3-glucanase gene resulted in higher cellulose, hemicellulose and pectin contents and a lower lignin content in the fibres. Increases in the uronic acid content in particular fractions (with the exception of the 1 M KOH-soluble fraction of hemicelluloses) and changes in the sugar composition of the cell wall were detected in the fibres of the transgenic flax when compared to the contents for the control plants. The callose content was lower in the fibres of the transgenic flax. Additionally, the analysis of phenolic compound contents in five fractions of the cell wall revealed important changes, which were reflected in the antioxidant potential of these fractions. CONCLUSION: Overexpression of the ß-1,3-glucanase gene has a significant influence on the biochemical composition of flax fibres. The constitutive overproduction of ß-1,3-glucanase causes a decrease in the callose content, and the resulting excess glucose serves as a substrate for the production of other polysaccharides. The monosaccharide excess redirects the phenolic compounds to bind with polysaccharides instead of to partake in lignin synthesis. The mechanical properties of the transgenic fibres are strengthened by their improved biochemical composition, and the increased antioxidant potential of the fibres supports the potential use of transgenic flax fibres for biomedical applications.

Partial characterization of an exopolysaccharide secreted by a marine bacterium, Vibrio neocaledonicus sp. nov., from New Caledonia.

AIMS: Exopolysaccharides (EPS) are industrially valuable molecules with numerous useful properties. This study describes the techniques used for the identification of a novel Vibrio bacterium and preliminary characterization of its EPS. METHODS AND RESULTS: Bioprospection in marine intertidal areas of New Caledonia followed by screening for EPS producing brought to selection of the isolate NC470. Phylogenetic analysis (biochemical tests, gene sequencing and DNA-DNA relatedness) permitted to identify NC470 as a new member of the Vibrio genus. The EPS was produced in batch fermentation, purified using the ultrafiltration process and analysed by colorimetry, Fourier Transform Infrared spectroscopy, gas chromatography, Nuclear Magnetic Resonance and HPLC-size exclusion chromatography. This EPS exhibits a high N-acetyl-hexosamines and uronic acid content with a low amount of neutral sugar. The molecular mass was 672 × 10(3)  Da. These data are relevant for possible technological exploitation. CONCLUSIONS: We propose the name Vibrio neocaledonicus sp. nov for this isolate NC470, producing an EPS with an unusual sugar composition. Comparison with other known polymers permitted to select applications for this polymer. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributes to evaluate the marine biodiversity of New Caledonia. It also highlights the biotechnological potential of New Caledonia marine bacteria.

Comparative studies on chemical parameters and antioxidant properties of stipes and caps of shiitake mushroom as affected by different drying methods.

BACKGROUND: Shiitake, the second most cultivated mushroom, is famous for its high nutritional value and medicinal properties. In this study, various chemical parameters and antioxidant properties of caps and stipes of shiitake mushroom dried by different methods (freeze-drying, shade drying and hot air drying) were comparatively investigated by spectrophotometric assays, high-performance liquid chromatography, 1,1'-diphenyl-2-picrylhydrazyl assay, ferric reducing power assay and lipid peroxidation inhibition assay. RESULTS: The contents of amino acids, neutral sugar and total phenolics in stipes were higher than those in caps of shiitake, while caps showed advantages in terms of their contents of protein and eritadenine. The chemical parameters and antioxidant activities of shiitake were significantly affected by the drying method used. CONCLUSION: The contents of total phenolics, amino acids and neutral sugar in stipes were higher than those in caps of shiitake, which suggested that stipes were more nutritional than caps in some respects. Hot air drying at 50 °C resulted in high total phenolic, amino acid, uronic acid and neutral sugar contents and antioxidant activities, which could be useful for the application of shiitake and related products in the food industry.

[Influence of different preparation on the content of carbohydrate and physicochemical properties of polysaccharides from raw paeoniae radix alba and stir-baked paeoniae radix alba].

OBJECTIVE: To observe the difference of two preparation methods on the content of carbohydrate and physicochemical properties of polysaccharides from raw Paeoniae Radix Alba and stir-baked Paeoniae Radix Alba. METHODS: Polysaccharides extracted from raw Paeoniae Radix Alba and stir-baked Paeoniae Radix Alba with water were precipitated by ethanol and named as BSEP-S and BSEP-C, respectively. In the same way, those deposited by acetone were named as BSAP-S and BSAP-C. Their physicochemical properties, including extraction yield, the content of carbohydrate, elemental composition and monosaccharide composition, were determined. RESULTS: Extraction yield, sugar content and uronic acid content of BSEP-S was 1.56%, 80.56% and 3.33% , BSEP-C 1.18%, 80.79% and 5.47%, BSAP-S 1.58%, 86.50% and 3.79%, and BSAP-C 1.54%, 81.64% and 3.25%, respectively. In addition, monosaccharide compositions showed that glucose was the main monosaccharide and successively reached 92.21%, 87.30%, 91.45% and 86.62%. CONCLUSION: Yield and content of carbohydrate from raw Paeoniae Radix Alba and stir-baked Paeoniae Radix Alba by water extraction-ethanol precipitation are a little higher than that by water extraction-acetone precipitation, but monosaccharide compositions are almost the same. Different preparation has significant impact on the yield and the content of carbohydrate in Paeoniae Radix Alba by stir-baked method, and it can decrease the dissolution of polysaccharide.

Mass spectrometric method for determining the uronic acid epimerization in heparan sulfate disaccharides generated using nitrous acid.

Heparan sulfate (HS) glycosaminoglycans (GAGs) regulate a host of biological functions. To better understand their biological roles, it is necessary to gain understanding about the structure of HS, which requires identification of the sulfation pattern as well as the uronic acid epimerization. In order to model HS structure, it is necessary to quantitatively profile depolymerization products. To date, liquid chromatography-mass spectrometry (LC-MS) methods for profiling heparin lyase decomposition products have been shown. These enzymes, however, destroy information about uronic acid epimerization. Deaminative cleavage using nitrous acid (HONO) is a classic method for GAG depolymerization that retains uronic acid epimerization. Several chromatographic methods have been used for analysis of deaminative cleavage products. The chromatographic methods have the disadvantage that there is no direct readout on the structures producing the observed peaks. This report demonstrates a porous graphitized carbon (PGC)-MS method for the quantification of HONO generated disaccharides to obtain information about the sulfation pattern and uronic acid epimerization. Here, we demonstrate the separation and identification of uronic acid epimers as well as geometric sulfation isomers. The results are comparable to those expected for benchmark HS and heparin samples. The data demonstrate the utility of PGC-MS for quantification of HS nitrous acid depolymerization products for structural analysis of HS and heparin.

Cell wall polysaccharides are involved in P-deficiency-induced Cd exclusion in Arabidopsis thaliana.

The physiological and molecular mechanisms leading to the competitive interactions between phosphorus (P) and metal elements are a matter of debate. In this study, we found that P deficiency can alleviate cadmium (Cd) toxicity in Arabidopsis thaliana (Col-0). Under P deficiency (-P), less Cd was accumulated in the plants and the root cell walls, indicating the operation of a P-deficiency-induced Cd exclusion mechanism. However, organic acid efflux was similar under -P+Cd and +Cd treatments, suggesting that organic acid efflux is not responsible for the Cd exclusion. Interestingly, P deficiency significantly decreased cell wall polysaccharides (pectin and hemicellulose) contents and pectin methylesterase activity, and decreased the Cd retained by the extracted root cell wall. Therefore, we conclude that the modification of cell wall composition is responsible for the Cd exclusion of the root under P deficiency.

Predictive models of biohydrogen and biomethane production based on the compositional and structural features of lignocellulosic materials.

In an integrated biorefinery concept, biological hydrogen and methane production from lignocellulosic substrates appears to be one of the most promising alternatives to produce energy from renewable sources. However, lignocellulosic substrates present compositional and structural features that can limit their conversion into biohydrogen and methane. In this study, biohydrogen and methane potentials of 20 lignocellulosic residues were evaluated. Compositional (lignin, cellulose, hemicelluloses, total uronic acids, proteins, and soluble sugars) as well as structural features (crystallinity) were determined for each substrate. Two predictive partial least square (PLS) models were built to determine which compositional and structural parameters affected biohydrogen or methane production from lignocellulosic substrates, among proteins, total uronic acids, soluble sugars, crystalline cellulose, amorphous holocelluloses, and lignin. Only soluble sugars had a significant positive effect on biohydrogen production. Besides, methane potentials correlated negatively to the lignin contents and, to a lower extent, crystalline cellulose showed also a negative impact, whereas soluble sugars, proteins, and amorphous hemicelluloses showed a positive impact. These findings will help to develop further pretreatment strategies for enhancing both biohydrogen and methane production.

Penicillium solitum produces a polygalacturonase isozyme in decayed Anjou pear fruit capable of macerating host tissue in vitro.

A polygalacturonase (PG) isozyme was isolated from Penicillium solitum-decayed Anjou pear fruit and purified to homogeneity with a multistep process. Both gel filtration and cation exchange chromatography revealed a single PG activity peak, and analysis of the purified protein showed a single band with a molecular mass of 43 kDa, which is of fungal origin. The purified enzyme was active from pH 3.5-6, with an optimum at pH 4.5. PG activity was detectable 0-70 C with 50 C maximum. The purified isozyme was inhibited by the divalent cations Ca(2+), Mg(2+), Mn(2+) and Fe(2+) and analysis of enzymatic hydrolysis products revealed polygalacturonic acid monomers and oligomers. The purified enzyme has an isoelectric point of 5.3 and is not associated with a glycosylated protein. The PG isozyme macerated fruit tissue plugs in vitro and produced ~1.2-fold more soluble polyuronides from pear than from apple tissue, which further substantiates the role of PG in postharvest decay. Data from this study show for the first time that the purified PG produced in decayed Anjou pear by P. solitum, a weakly virulent fungus, is different from that PG produced by the same fungus in decayed apple.