De-esterification Of Pectin Research Articles

Rootstock effects on bitter pit incidence in 'Honeycrisp' apples are associated with changes in fruit's cell wall chemical properties.

Bitter pit (BP) is a physiological disorder of apples that often appears during or after cold storage. Despite being defined as a calcium deficiency disorder, BP is a complex process that is not only affected by the total Ca2+ content in the fruit but also by the proper cellular Ca2+ homeostasis and partitioning. Early investigations have also suggested that rootstocks could affect BP development and severity. In the present study, rootstock effects on BP development were assessed on ‘Honeycrisp’ trees that were grafted on 14 different rootstocks (B.10, G.11, G.202, G.214, G.30, G.41, G.935, G.969, M.26 EMLA, M.9, V.1, V.5, V.6, and V.7). We evaluated BP incidence at harvest, and three months after cold storage for four, and three growing seasons, respectively. BP incidence was significantly reduced in ‘Honeycrisp’ trees on B.10 compared to other rootstocks, whereas trees on V.6 showed the highest percentage of BP at harvest and after cold storage. ‘Honeycrisp’ apples were collected from three different rootstocks (B.10, G.41, and V.6) two months after cold storage and evaluated for mineral nutrient composition, Ca2+ homeostasis, and cell wall properties, e.g., pectin content, pectin de-esterification rate and pectin methylesterase (PME) activity. Water-soluble and insoluble pectin content was markedly higher in fruits from B.10 than in G.41 and V.6. We also observed increased PME enzyme activity and a greater degree of water-insoluble pectin de-esterification in ‘Honeycrisp’ apples from V.6 compared to those from B.10. A significantly higher Ca2+ was found in the fruits from B.10 than G.41 and V.6. Higher Ca2+ and lower Mg2+ levels were also observed in the cell wall and water-insoluble pectin fractions of the fruits from B.10 compared to G.41 and V.6. However, the ratio of cell wall-bound Ca2+ to total Ca2+ was lower in B.10 compared to G.41 and V.6. Together, our results indicate that the tolerance of B.10 to BP could be attributed to a reduced PME activity and lower pectin de-esterification level, which in turn reduced the amount of Ca2+ cross-linked with pectin, and probably increased the apoplastic free calcium concentrations that is essential for maintaining cell membrane integrity and reducing BP development.

Genome-wide identification of PME gene family and expression of candidate genes associated with aluminum tolerance in tea plant (Camellia sinensis)

BackgroundThe major aluminum (Al) detoxication mechanism of tea plant (Camellia sinensis), as an Al hyperaccumulator plant, is the fixation of almost 70% of Al in the cell walls. Pectin is the primary constituent of cell walls, a degree of methylation of pectin polysaccharides regulated by the pectin methylesterase (PME) genes can greatly affect the Al binding capacity. The knowledge on PME gene family in tea plant is still poor.ResultsWe identified 66 (CsPME1-CsPME66) PME genes from C. sinensis genome. We studied their protein characterization, conserved motifs, gene structure, systematic evolution and gene expression under Al treatments, to establish a basis for in-depth research on the function of PMEs in tea plant. Gene structures analysis revealed that the majority of PME genes had 2–4 exons. Phylogenetic results pointed out that the PME genes from the same species displayed comparatively high sequence consistency and genetic similarity. Selective pressure investigation suggested that the Ka/Ks value for homologous genes of PME family was less than one. The expression of CsPMEs under three Al concentration treatments was tissue specific, eight PME genes in leaves and 15 in roots displayed a trend similar to of the Al contents and PME activities under Al concentration treatments, indicating that the degree of pectin de-esterification regulated by PME was crucial for Al tolerance of tea plant.ConclusionsSixty-six CsPME genes were identified for the first time in tea plant. The genome-wide identification, classification, evolutionary and transcription analyses of the PME gene family provided a new direction for further research on the function of PME gene in Al tolerance of tea plant.

Effect of Sterilization on the Degree of Esterification, FTIR Analysis, and Antibacterial Activity of Durian-Rind Pectin

Pectin is a common food ingredient used as a rheology modifier and recently recognized as an emerging bioactive compound. Degree of esterification (DE) and molecular weight (MW) are important determinants of its bioactivity. This study evaluated the effect of moist heat sterilization (121 °C, 15 min) on pectin from Indonesian durian rind as an alternative method to modify pectin. Sterilized pectin was compared in terms of DE, Fourier transform infrared (FTIR) analysis, gel-forming ability, and antibacterial activity to non-sterilized pectin and standard citrus-peel pectin. Durian-rind pectin was identified as a low-methoxyl pectin with DE of 26.50% and weak antibacterial activity. After sterilization, the DE and pH decreased. It lost the ability to form a gel which indicated pectin was degraded to lower molecules. Loss of bands at 1760-1745 cm-1 indicated that pectin underwent ester hydrolysis and generated free carboxyl groups. On the other hand, the sterilized durian-rind pectin showed strong antibacterial activity towards Staphylococcus aureus and Escherichia coli, with a reduction of 5 log cycles and 3 log cycles, respectively (with the initial bacterial level of 5 log cfu/mL). These results indicated that depolymerization and deesterification of pectin by heat sterilization was able to improve the antibacterial activity of durian-rind pectin.

Screening of the proteins related to the cultivar-dependent cadmium accumulation of Brassica parachinensis L.

Cultivar-dependent cadmium (Cd) accumulation was principal in developing Cd-pollution safe cultivars (PSCs). Proteins related to different Cd accumulations of the low-Cd-accumulating (SJ19) and high-Cd-accumulating (CX4) cultivars were investigated by iTRAQ analysis. Higher Cd bioaccumulation factors and translocation factor in CX4 than in SJ19 were consistent with the cultivar-dependent Cd accumulations. The Cd uptake was promoted in CX4 due to its higher expression of Cd-binding proteins and the lower expression of Cd-efflux proteins in roots. What's more, significantly elevated thiol groups (PC2 and PC3) in CX4 under Cd stress might contribute to the high Cd accumulation in roots and the root-to-shoot translocation of Cd-PC complex. Up-regulated proteins involved in cellulose biosynthesis and pectin de-esterification in SJ19 enhanced the Cd sequestration of root cell walls, which was considered as the predominant strategy for reducing Cd accumulation in shoots. The present study provided novel insights in the cultivar-dependent Cd accumulation in shoots of B. parachinensis.

Comparative study on gelling properties of low methoxyl pectin prepared by high hydrostatic pressure-assisted enzymatic, atmospheric enzymatic, and alkaline de-esterification

The gel behavior of low methoxyl pectin (LMP) de-esterified by high hydrostatic pressure-assisted enzymatic (HHP-pectin, 400 MPa/40 °C/12 min), atmospheric enzymatic (E-pectin, 40 °C/2 h), and alkaline method (A-pectin, pH = 12/25 °C/25 min) was comparatively investigated, with regard to gel strength, water-holding capacity (WHC), rheological properties, and microstructure of pectin gels. Results showed that the de-esterification method had a strong influence on gelling properties of LMPs. The gelation of all LMPs was significantly affected by pectin, Ca2+ concentration and pH of the solution, but enzymatically de-esterified LMPs formed gels with higher gel strength, WHC and better viscoelasticity than those from alkaline de-esterified method. HHP-pectin gels showed even higher gel strength at the same Ca2+ concentrations and better WHC at the same pH, as compared to those from E-pectin. Moreover, the de-esterification time for HHP-pectin was much shorter than that for E-pectin. These findings indicated that HHP could be a potential alternative for high efficient pectin de-esterification, preparing LMPs with better gelling property.

Physiochemical, rheological and emulsifying properties of low methoxyl pectin prepared by high hydrostatic pressure-assisted enzymatic, conventional enzymatic, and alkaline de-esterification: A comparison study

Abstract: Low methoxyl pectin (LMP) was prepared by high hydrostatic pressure assisted enzymatic treatment (HHP-pectin). The physicochemical properties, rheological characteristics and emulsifying characteristics of LMP prepared in this manner were compared the characteristics of LMPs de-esterified using the conventional enzymatic (E-pectin) and alkaline (A-pectin) methods. The obtained results showed that under the optimal HHP conditions, the degree of methoxylation (DM) of pectin decreased from 79.91% to 37.02% within 12 min, whereas 120 min and 20 min, respectively, were required to obtain a similar DM value for the atmospheric enzymatic and alkaline treatments. Moreover, the de-esterification method had a strong influence on rheological and emulsifying properties. Compared to A-pectin, HHP-pectin and E-pectin exhibited higher viscosity, accompanied by larger average molecular weight, particle size, more negative zeta-potential and higher activation energy. All of the emulsions exhibited good stability at acidic conditions with pH 3. While the emulsion prepared using A-pectin showed the smallest particle mean diameters and better centrifugation stability, the emulsion prepared using HHP-pectin showed the best long-term emulsifying stability during 21-day storage. These results clearly demonstrated that HHP-assisted enzymatic method is a more efficient, rapid and eco-friendly alternative for pectin de-esterification, particularly for preparing LMPs with better thickening and stable emulsifying properties.

Effect of alkaline de-esterified pectin on the complex coacervation with pea protein isolate under different mixing conditions

High methoxy citrus pectin (UM88) was saponified to produce modified pectin [M(72, 42, and 9)], with different levels of degree of esterification (DE), to investigate the complex coacervation of pea protein isolate (PPI) with pectin [UM88 and M(72, 42, and 9)]. Regardless of the DE value of pectin, the critical pH corresponding to when insoluble complexes form shifted to higher pH as the mixing ratio increased. The maximum amount of coacervates formed at a biopolymer-mixing ratio of 8:1, 8:1, 10:1 and 15:1 for PPI with UM88, M72, M42, and M9, respectively. Maximum interactions for the protein-pectin admixtures occurred between pH 3.70 and 3.85. PPI complexed with modified pectin displayed greater interactions under their optimal mixing conditions compared to the unmodified pectin. The de-esterification of pectin resulted in more rigid and stiffer pectin, which enhanced its interaction with PPI by shifting the critical parameters to a higher value.

Pectin De-methylesterification and AGP Increase Promote Cell Wall Remodeling and Are Required During Somatic Embryogenesis of Quercus suber.

Somatic embryogenesis is a reliable system for in vitro plant regeneration, with biotechnological applications in trees, but the regulating mechanisms are largely unknown. Changes in cell wall mechanics controlled by methylesterification of pectins, mediated by pectin methylesterases (PMEs) and pectin methyl esterase inhibitors (PMEIs) underlie many developmental processes. Arabinogalactan proteins (AGPs) are highly glycosylated proteins located at the surface of plasma membranes, in cell walls, and in extracellular secretions, with key roles in a range of different processes. In this study, we have investigated changes in two cell wall components, pectins and AGPs, during somatic embryogenesis in Quercus suber, a forest tree of high economic and ecologic value. At early embryogenesis stages, cells of proembryogenic masses showed high levels of esterified pectins and expression of QsPME and QsPMEI genes encoding a PME and a putative PMEI, respectively. At advanced stages, differentiating cells of heart, torpedo and cotyledonary embryos exhibited walls rich in de-esterified pectins, while QsPME gene expression and PME activity progressively increased. AGPs were detected in cell walls of proembryogenic masses and somatic embryos. QsLys-rich-AGP18, QsLys-rich-AGP17, and QsAGP16L1 gene expression increased with embryogenesis progression, as did the level of total AGPs, detected by dot blot with β-glucosyl Yariv reagent. Immuno dot blot, immunofluorescence assays and confocal analysis using monoclonal antibodies to high- (JIM7, LM20) and low- (JIM5, LM19) methylesterified pectins, and to certain AGP epitopes (LM6, LM2) showed changes in the amount and distribution pattern of esterified/de-esterified pectins and AGP epitopes, that were associated with proliferation and differentiation and correlated with expression of the PME and AGP genes analyzed. Pharmacological treatments with catechin, an inhibitor of PME activity, and Yariv reagent, which blocks AGPs, impaired the progression of embryogenesis, with pectin de-esterification and an increase in AGP levels being necessary for embryo development. Findings indicate a role for pectins and AGPs during somatic embryogenesis of cork oak, promoting the cell wall remodeling during the process. They also provide new insights into the regulating mechanisms of somatic embryogenesis in woody species, for which information is still scarce, opening up new possibilities to improve in vitro embryo production in tree breeding.

Biochemical Reconstruction of a Metabolic Pathway from a Marine Bacterium Reveals Its Mechanism of Pectin Depolymerization.

Pectin is a complex uronic acid-containing polysaccharide typically found in plant cell walls, though forms of pectin are also found in marine diatoms and seagrasses. Genetic loci that target pectin have recently been identified in two phyla of marine bacteria. These loci appear to encode a pectin saccharification pathway that is distinct from the canonical pathway typically associated with phytopathogenic terrestrial bacteria. However, very few components of the marine pectin metabolism pathway have been experimentally validated. Here, we biochemically reconstructed the pectin saccharification pathway from a marine Pseudoalteromonas sp. in vitro and show that it results in the production of galacturonate and the key metabolic intermediate 5-keto-4-deoxyuronate (DKI). We demonstrate the sequential de-esterification and depolymerization of pectin into oligosaccharides and the synergistic action of glycoside hydrolases (GHs) to fully degrade these oligosaccharides into monosaccharides. Furthermore, we show that this pathway relies on enzymes belonging to GH family 105 to carry out the equivalent chemistry afforded by an exolytic polysaccharide lyase (PL) and KdgF in the canonical pectin pathway. Finally, we synthesize our findings into a model of marine pectin degradation and compare it with the canonical pathway. Our results underline the shifting view of pectin as a solely terrestrial polysaccharide and highlight the importance of marine pectin as a carbon source for suitably adapted marine heterotrophs. This alternate pathway has the potential to be exploited in the growing field of biofuel production from plant waste.IMPORTANCE Marine polysaccharides, found in the cell walls of seaweeds and other marine macrophytes, represent a vast sink of photosynthetically fixed carbon. As such, their breakdown by marine microbes contributes significantly to global carbon cycling. Pectin is an abundant polysaccharide found in the cell walls of terrestrial plants, but it has recently been reported that some marine bacteria possess the genetic capacity to degrade it. In this study, we biochemically characterized seven key enzymes from a marine bacterium that, together, fully degrade the backbone of pectin into its constituent monosaccharides. Our findings highlight the importance of pectin as a marine carbon source available to bacteria that possess this pathway. The characterized enzymes also have the potential to be utilized in the production of biofuels from plant waste.

Pectin modification assisted by nitrogen glow discharge plasma

This study is concerned with modification of high methoxy pectin (HMP) assisted by nitrogen AC glow discharge plasma. Intrinsic viscosity of plasma treated pectin (PTP) was measured as primary index for structural changes in pectin molecules at different plasma conditions. The intrinsic viscosity of pectin increased within 7 min of exposure to plasma treatment and then remained constant. According to Fourier Transform Infrared Spectrometry (FT-IR) analysis, the intensity of carboxylate peaks increased in PTP samples due to de-esterification of pectin. Moreover, by decreasing the degree of esterification (DE) in PTP samples, the results of FT-IR were confirmed. Based on high performance size-exclusion chromatography (HPSEC) analysis, the molecular weight of PTP reduced. PTP gel had higher storage and loss modulus and shorter linear viscoelastic region. Moreover, X-ray diffraction (XRD) studies indicated an increase of crystallinity in PTP due to the formation of hydrogen bonds. Results revealed that nitrogen AC glow discharge plasma had significant influence on physicochemical and functional properties of pectin. The result of this study showed AC glow discharge plasma has promising potential towards modification of pectin.

Characterization and Expression Pattern of a Putative Pectin Methylesterase Gene, BcMF27, in Brassica campestris ssp. chinensis

Pectin methylesterases (PMEs) play an important role in modifying cell wall. PMEs catalyze the de-esterification of pectin, an important compound of cell wall, to affect fertility in plant reproduction. However, little especially molecular mechanism about pectin methylesterase is studied in recent years despite its importance to reproductive development in flower plant. Here the bioinformatics analysis of BcMF27 (Brassica campestris Male Fertility 27) (BRAD: Bra000541 GenBank: KT600012) sequence isolated from Brassica campestris L. ssp. chinensis showed its highly and characteristically conserved structure as a pectin methylesterase. Transient expression analysis in the onion epidermal cells revealed the product of BcMF27 was a transmembrane protein. Real-time RT-PCR and in situ hybridization suggested that BcMF27 was expressed in pollen grain and pollen tube. This study demonstrates that BcMF27 encodes a transmembrane pollen- and pollen tube-specific PME gene, and is also considered to help further understand the biological function of pectin methylesterases and the molecular mechanism of pollen development, pollen tube growth as a genic tool.

Properties Of Two Novel Esterases Identified From Culture Supernatant Of Penicillium Purpurogenum Grown On Sugar Beet Pulp

The filamentous fungus Penicillium purpurogenum grows on a variety of natural carbon sources, such as sugar beet pulp, and secretes to the medium a large number of enzymes that degrade the carbohydrate components of lignocellulose. Sugar beet pulp is rich in pectin, and the purpose of this work is to identify novel esterases produced by the fungus, which may participate in pectin degradation. Partially purified culture supernatants of the fungus grown on sugar beet pulp were subjected to mass spectrometry analysis. Peptides thus identified, which may be part of potential esterases were probed against the proteins deduced from the fungal genome sequence. The cDNAs of two putative esterases identified were expressed in Pichia pastoris and their properties studied. One of these enzymes, named FAET, is a feruloyl esterase, while the other, PE, is classified as a pectin methyl esterase. These findings add to our knowledge of the enzymology of pectin degradation by Penicillium purpurogenum, and define properties of two novel esterases acting on de-esterification of pectin. Their availability may be useful as tools for the study of pectin structure and degradation.

Evolutionary origins of pectin methylesterase genes associated with novel aspects of angiosperm pollen tube walls

The early evolution of angiosperms was marked by a number of innovations of the reproductive cycle including an accelerated fertilization process involving faster transport of sperm to the egg via a pollen tube. Fast pollen tube growth rates in angiosperms are accompanied by a hard shank-soft tip pollen tube morphology. A critical actor in that morphology is the wall-embedded enzyme pectin methylesterase (PME), which in type II PMEs is accompanied by a co-transcribed inhibitor, PMEI. PMEs convert the esterified pectic tip wall to a stiffer state in the subapical flank by pectin de-esterification. It is hypothesized that rapid and precise targeting of PME activity was gained with the origin of type II genes, which are derived and have only expanded since the origin of vascular plants. Pollen-active PMEs have yet to be reported in early-divergent angiosperms or gymnosperms. Gene expression studies in Nymphaea odorata found transcripts from four type II VGD1-like and 16 type I AtPPME1-like homologs that were more abundant in pollen and pollen tubes than in vegetative tissues. The near full-length coding sequence of one type II PME (NoPMEII-1) included at least one PMEI domain. The identification of possible VGD1 homologs in an early-diverging angiosperm suggests that the refined control of PMEs that mediate de-esterification of pectins near pollen tube tips is a conserved feature across angiosperms. The recruitment of type II PMEs into a pollen tube elongation role in angiosperms may represent a key evolutionary step in the development of faster growing pollen tubes.

BnPME is progressively induced after microspore reprogramming to embryogenesis, correlating with pectin de-esterification and cell differentiation in Brassica napus.

BackgroundPectins are one of the main components of plant cell walls. They are secreted to the wall as highly methylesterified forms that can be de-esterified by pectin methylesterases (PMEs). The degree of methylesterification of pectins changes during development, PMEs are involved in the cell wall remodeling that occurs during diverse plant developmental processes. Nevertheless, the functional meaning of pectin-related wall remodeling in different cell types and processes remains unclear. In vivo, the microspore follows the gametophytic pathway and differentiates to form the pollen grain. In vitro, the microspore can be reprogrammed by stress treatments becoming a totipotent cell that starts to proliferate and follows the embryogenic pathway, a process known as microspore embryogenesis.ResultsTo investigate if the change of developmental programme of the microspore towards embryogenesis involves changes in pectin esterification levels, which would cause the cell wall remodeling during the process, in the present study, dynamics of PME expression and degrees of pectin esterification have been analysed during microspore embryogenesis and compared with the gametophytic development, in Brassica napus. A multidisciplinary approach has been adopted including BnPME gene expression analysis by quantitative RT-PCR, fluorescence in situ hybridization, immuno-dot-blot and immunofluorescence with JIM5 and JIM7 antibodies to reveal low and highly-methylesterified pectins. The results showed that cell differentiation at advanced developmental stages involved induction of BnPME expression and pectin de-esterification, processes that were also detected in zygotic embryos, providing additional evidence that microspore embryogenesis mimics zygotic embryogenesis. By contrast, early microspore embryogenesis, totipotency and proliferation were associated with low expression of BnPME and high levels of esterified pectins.ConclusionsThe results show that the change of developmental programme of the microspore involves changes in pectin esterification associated with proliferation and differentiation events, which may cause the cell wall remodeling during the process. The findings indicate pectin-related modifications in the cell wall during microspore embryogenesis, providing new insights into the role of pectin esterification and cell wall configuration in microspore totipotency, embryogenesis induction and progression.

Modified pectin from Theobroma cacao induces potent pro-inflammatory activity in murine peritoneal macrophage

In vitro effects of acetylated pectin (OP) isolated from cacao pod husks (Theobroma cacao L.), its partially deacetylated and de-esterified form (MOP), and a commercial homogalacturonan (PG) were investigated on murine peritoneal macrophages. MOP stood out among the studied pectins. After 48h of incubation, compared with the control group, it was able to promote significant macrophage morphological differentiation from resident to activated stage and also stimulated nitric oxide production, which reached a level of 85% of that of LPS stimulus. In the presence of the highest tested concentration of MOP (200μg·mL⿿1), the levels of the cytokines TNF-α (6h) and IL-12 and IL-10 (48h) increased substantially in relation to untreated cells. Our results show that the partial deacetylation and de-esterification of pectin extracted from cacao pod husks (T. cacao L.) produced a polymer with greater ability than its native form to activate macrophages to a cytotoxic phenotype. Like this, they provide the possibility of a therapeutic application to MOP, which could lead to a decreased susceptibility to microbial infection besides antitumor activity. Additionally, the present results also corroborate with the proposition of that the chemical modifications of the biopolymers can result in an improved molecule with new possibilities of application.

Pectins esterification in the apoplast of aluminum-treated pea root nodules

Aiming to elucidate the possible involvement of pectins in aluminum-mediated growth inhibition the distribution of pectins in the apoplast of root nodules was investigated. Experiments were performed on the pea (Pisum sativum L.) root nodules treated with aluminum (50μM AlCl3, for 2 or 24h). For histochemical acidic pectin localization we used ruthenium red staining. Immunolabeling techniques with monoclonal antibodies specific to high methyl-esterified pectin (JIM7), low methyl-esterified pectin (JIM5) and calcium cross-linked pectin (2F4) were used to re-examine the pattern of pectin esterification and distribution. After immunolabeling the samples were observed using a fluorescent and transmission electron microscope.Ruthenium red staining showed that acid pectin content increased in the apoplast of Al-treated nodules and immunolocalization of pectin epitopes revealed that the fraction of de-esterified pectins increased significantly under Al stress. JIM5 and 2F4 epitopes were located on the inner surface of the primary cell wall with higher intensity at cell corners lining the intercellular spaces and at infection threads (ITs) walls. By contrast, JIM 7 labels all walls uniformly throughout the nodule. In the presence of Al, the increase of JIM5 and 2F4 labeling in thick plant and IT walls, together with a decrease of JIM7 labeling was observed.These results indicate a specific role for pectin de-esterification in the process of wall thickening and growth inhibition. In particular, Al-dependent increase in pectin content and their low methyl esterification degree correlate with wall thickness and higher rigidity, and in this way, may affect IT and nodules growth.

Research of technological properties of biochemically modified pectin substances

The paper presents the results of experimental studies to determine the mass fraction of effectors – calcium ions, which allow you to adjust the solubility of low-esterified pectin gel to create structural compositions depending on the time and concentration of enzymatic de-esterification of pectin in the solution. Injection of calcium salts in a mixture having influence on both the solubility and the viscosity characteristics and the gelling ability of modified pectin solution. With injection of calcium salts in solution of citrus pectin it is observed their different behavior depending on the nature of amount of salts and introduced pectin. With an increase in the mass fraction of low-esterified pectin and calcium salts take place a gradual increase in viscosity of the solution with a transition to the dense gel, but higher concentrations of calcium lead to syneresis process. It has been established that structural and mechanical properties of the gel affect the amount and type of pectin, as well as in its content in the composition of calcium ions. It is shown that low-esterified pectin substances may make up the thermoreversible gels. Anions of calcium salts significantly affect the structuring of the pectin and calcium cation allows receiving a variety of gel compositions with a mass fraction of pectin is not less than 0,6 g per 100 cm 3 , which is important for producing structured and canned foods.

Physicochemical properties of modified citrus pectins extracted from orange pomace.

Modified pectin is a polysaccharide rich in galacturonic acid altered by pH adjustment and thermal treatment used especially as an anti-cancer agent. The aim of this work was to study the physical and chemical properties of modified pectins extracted from orange pomace with citric and nitric acids. The galacturonic acid content, degree of esterification, Fourier Transform Infrared Spectroscopy profile, molecular weight, intrinsic viscosity, rheological properties and antioxidant activity of the pectins were evaluated. The modification process caused the de-esterification of pectins and a decrease of molecular weight due to removal of neutral sugars, maintaining the linear chain of galacturonic acid. Such changes also caused a significant increase in the in vitro antioxidant activity (p ≤ 0.05) and influenced the rheological properties of pectin, reducing its viscosity. This work showed that the modification of pectin from orange pomace with citric and nitric acids altered its structural and physical characteristics as well as its biological activity toward a free-radical.

Pectin Methylesterase of Datura species, purification, and characterization from Datura stramoniumand its application

Pectin methylesterases (PME; EC 3.1.1.11) involved in de-esterification of pectin and have applicability in food, textiles, wines, pulp, and paper industries. In the present study, we compared PME activity of different parts of 3 Datura species and found that fruit coat showed maximum PME activity followed by leaf and seed. PME from leaves of D. stramonium (DsPME) was purified and characterized. DsPME showed optimum activity at 60 °C and pH 9 in the presence of 0.3 M NaCl. DsPME was stable at 70 °C and retained more than 40% activity after 60 min of incubation. However, enzyme activity completely abolished at 80 after 5 min of incubation. It follows Michaelis-Menten enzyme kinetics. Km and Vmax with citrus pectin were 0.008 mg/ml and 16.96 µmol/min, respectively. DsPME in combination with polygalactourenase (PGA) increased the clarity of orange, apple, pomegranate and pineapple juices by 2.9, 2.6, 2.3, and 3.6 fold, respectively in comparison to PGA alone. Due to very high de-esterification activity, easy denaturation and significant efficacy in incrementing clarification of fruit juice makes DsPME useful for industrial application.

Shading affects flesh calcium uptake and concentration, bitter pit incidence and other fruit traits in “Greensleeves” apple

Bitter pit (BP) is a physiological disorder in apple (Malus×domestica Borkh) believed to be triggered by low Ca2+ concentrations in the fruit, and which may be influenced by environmental conditions. The objectives of this study were to explore the effects of tree shading on total and cell wall bound fruit Ca2+ content and fruit susceptibility to BP. ‘Greensleeves’ (GS) apple trees were cultivated under shaded and non-shaded conditions. The shading condition was applied 70 days after full bloom by reducing about 50% of the sunlight reaching the tree canopies. Shading increased stem water potential and leaf stomatal conductance. Bitter pit was observed only in shaded fruit. Cortical Ca2+ was most abundant in shaded fruit without BP symptoms, intermediary in shaded fruit with BP, and lowest in non-shaded fruit. The cell wall Ca2+ concentration was higher in shaded than non-shaded fruit, but shaded fruit with and without BP had similar cell wall Ca2+ concentrations. The degree of pectin deesterification and the expression of two pectin methylesterases (PME1 and PME2) were higher in shaded fruit than in non-shaded fruit. The percentage of total cortical Ca2+ bound to the cell wall was highest in shaded fruit with BP. Shaded fruit without BP had lower Mg2+ content and Mg2+/Ca2+ ratio in cortical tissue than shaded fruit with BP. The results indicate that although shade increases fruit Ca2+ uptake, it also enhances fruit susceptibility to BP by increasing Mg2+ uptake and Ca2+ binding to the cell wall in fruit cortical tissue.