Influence of enzymatic and acidic demethoxylation on structure formation in sugar containing citrus pectin gels

Aim of the present study was to investigate the impact of different demethoxylation methods and the co-occuring side effects on the molecular properties and structure formation in pectin gels. A high-methoxylated citrus pectin (HMP) was demethoxylated using either hydrochloric acid or pectin methylesterases of plant (pPME) or fungal (fPME) origin. pPME treatment causes a more block-wise distribution of free carboxyl groups, fPME or acidic treatment a random distribution. Twelve pectin samples with four different degrees of methoxylation (DM) between 62% and 41% were prepared. The gelation process was studied by oscillatory measurements. In pectin samples from pPME treatment structure formation started at higher temperature and the final gels were more elastic in comparison to pectin from the two other modifications. The impact of the block-wise distribution of the free carboxyl groups became more evident with decreasing DM. The gelling process of pectin samples with random distribution was similar independent of DM.Side effects of all demethoxylation reactions were an altered sodium ion content (high in enzymatically treated pectin, close to zero in acidic treated) and a decrease of the molecular weight with increasing degree of demethoxylation. These side effects additionally altered the gelation process and the final gel properties in different ways

Characterisation of pectins extracted from different parts of Malaysian durian rinds

Durian rinds are a potential source of pectin. The rinds consist of the outer layer of the fruit which is semiwoody and thorny and the inner layer, which is creamy white. Pectin may be present in varying amounts in different parts of the rind. Previous studies only investigated pectin content from the inner part of the rinds. Therefore, it is essential to provide scientific data on the pectin yield from whole durian rinds (including the thorns). Particularly, this information can be useful when preparing durian rinds for pectin extraction

PECTOPLATE: the simultaneous phenotyping of pectin methylesterases, pectinases and oligogalacturonides in plants during biotic stresses

Degradation of pectin, a major component of plant cell wall, is important for fungal necrotrophs to achieve a successful infection. The activities of pectin methylesterases (PMEs) from both plants and pathogens and the degree and pattern of pectin methylesterification are critical for the outcome of plant–pathogen interaction. Partial degradation of pectin by pectin degrading enzymes releases oligogalacturonides (OGs), elicitors of plant defense responses. Few analytical techniques are available to monitor pectin methylesterification-modulating machineries and OGs produced during plant pathogen interaction. In the present study, ruthenium red is presented as useful dye to monitor both Botrytis cinerea mycelium growth and the induction of PME activity in plant tissue during fungal infection. Moreover a simple, inexpensive and sensitive method, named PECTOPLATE, is proposed that allows a simultaneous phenotyping of PME and pectinase activities expressed during pathogen infection and of pectinase potential in generating OGs. The results in the manuscript also indicate that PME inhibitors can be used in PECTOPLATE as a tool to discriminate the activities of plant PMEs from those of pathogen PMEs expressed during pathogenesis

Modeling of the break process to improve tomato paste production quality : a thesis presented in partial fulfilment for the degree of Master of Engineering in Bioprocess at Massey University

The pectic enzyme, Pectinmethylesterase (PE) and Polygalacturonase I and II (PGI and PGII), in the tomato fruit released after crushing during tomato processing reduce the viscosity of tomato paste by breaking down the insoluble pectin in the cell wall. To achieve higher viscosity tomato paste, the cold break (60-95°C) processes can be used to inactivate the pectic enzyme and to achieve higher viscosity tomato paste. The study of tomato solids and PG enzyme activity showed that the levels of insoluble solids, total solids, pectin, and °Brix in Ferry Morse tomatoes were independent of fruit ripeness. The amount of PG enzymes was high in orange and dark red tomatoes and the activity of PG enzymes increased as a function of ripeness, from green to dark red. In the dark red tomato, the inactivation of PG enzyme activity was required to retain the level of pectin. Cold break temperatures below 60°C can not inactivate the PG enzyme activity. The PG enzymes started to be denatured when the hot break temperature was above 65°C and be completely destroyed when the break temperature was above 80°C. A mathematical model of the break process was formulated and Matlab programme was used to predict the effect of break temperatures on the pectin and PG enzyme concentration of the tomato pulp in the break tank for any inputs of feed rate (the flow rate to the break tank), feed ripeness, and residence time. The model was used to demonstrate the understanding and the optimisation of break process performance. Longer residence time of dark red tomato pulp in the break tank can decrease pectin fraction residual and increase enzyme inactivation in the tank temperature range 40 to 60°C. The pectin fraction remaining increased when the tank temperature was above 60°C because of the inactivation of PG enzymes. At 80°C there was no effect of residence time, the pectin fraction residual increased and reached 90% and enzyme fraction residual decreased to 10%. The effect of mixed tomato ripeness between the ripe fruit (orange and dark red) with the unripe fruit (green, breaker, and turning), the level of PG enzymes in the break tank decreased and affected on the higher pectin fraction remaining. Lower break temperature can be therefore used in this process to inactivate the low amount of PG enzyme and to achieve the same extent of pectin hydrolysis. The interruption of the feed coming into the break tank during tomato processing can increase the pectin fraction remaining and the enzyme fraction remaining in a new steady state when the feed was turned on

Spatial structure and composition of polysaccharide-protein complexes from Small Angle Neutron Scattering

We use Small Angle Neutron Scattering (SANS), with an original analysis method, to obtain both the characteristic sizes and the inner composition of lysozyme-pectin complexes depending on the charge density. Lysozyme is a globular protein and pectin a natural anionic semiflexible polysaccharide with a degree of methylation (DM) 0, 43 and 74. For our experimental conditions (buffer ionic strength I = 2.5 10-2 mol/L and pH between 3 and 7), the electrostatic charge of lysozyme is always positive (from 8 to 17 depending on pH). The pectin charge per elementary chain segment is negative and can be varied from almost zero to one through the change of DM and pH. The weight molar ratio of lysozyme on pectin monomers is kept constant. The ratio of negative charge content per volume to positive charge content per volume, -/+, is varied between 10 and 0.007. On a local scale, for all charged pectins, a correlation peak appears at 0.2 {\AA}-1 due to proteins clustering inside the complexes. On a large scale, the complexes appear as formed of spherical globules with a well defined radius of 10 to 50 nm, containing a few thousands proteins. The volume fraction Phi of organic matter within the globules derived from SANS absolute cross-sections is around 0.1. The protein stacking, which occurs inside the globules, is enhanced when pectin is more charged, due to pH or DM. The linear charge density of the pectin determines the size of the globules for pectin chains of comparable molecular weights whether it is controlled by the pH or the DM. The radius of the globules varies between 10 nm and 50 nm. In conclusion the structure is driven by electrostatic interactions and not by hydrophobic interactions. The molecular weight also has a large influence on the structure of the complexes since long chains tend to form larger globules. This maybe one reason why DM and pH are not completely equivalent in our system since DM 0 has a short mass, but this may not be the only one. For very low pectin charge (-/+ = 0.07), globules do not appear and the scattering signals a gel-like structure. We did not observe any beads-on-a-string structure

Improving the in vitro bioaccessibility of β-carotene using pectin added nanoemulsions

The intestinal absorption of lipophilic compounds such as β-carotene has been reported to increase when they are incorporated in emulsion-based delivery systems. Moreover, the reduction of emulsions particle size and the addition of biopolymers in the systems seems to play an important role in the emulsion properties but also in their behavior under gastrointestinal conditions and the absorption of the encapsulated compound in the intestine. Hence, the present study aimed to evaluate the effect of pectin addition (0%, 1%, and 2%) on the physicochemical stability of oil-in-water nanoemulsions containing β-carotene during 35 days at 4 °C, the oil digestibility and the compound bioaccessibility. The results showed that nanoemulsions presented greater stability and lower β-carotene degradation over time in comparison with coarse emulsion, which was further reduced with the addition of pectin. Moreover, nanoemulsions presented a faster digestibility irrespective of the pectin concentration used and a higher β-carotene bioaccessibility as the pectin concentration increased, being the maximum of ≈36% in nanoemulsion with 2% of pectin. These results highlight the potential of adding pectin to β-carotene nanoemulsions to enhance their functionality by efficiently preventing the compound degradation and increasing the in vitro bioaccessibility.This work was funded by the project AGL2015-65975-R (FEDER, MINECO, UE) and project RTI2018-094268-B-C21 (MCIU, AEI; FEDER, UE)

Activity and Process Stability of Purified Green Pepper (Capsicum annuum) Pectin Methylesterase

Pectin methylesterase (PME) from green bell peppers (Capsicum annuum) was extracted and purified by affinity chromatography on a CNBr-Sepharose-PMEI column. A single protein peak with pectin methylesterase activity was observed. For the pepper PME, a biochemical characterization in terms of molar mass (MM), isoelectric points (pI), and kinetic parameters for activity and thermostability was performed. The optimum pH for PME activity at 22 °C was 7.5, and its optimum temperature at neutral pH was between 52.5 and 55.0 °C. The purified pepper PME required the presence of 0.13 M NaCl for optimum activity. Isothermal inactivation of purified pepper PME in 20 mM Tris buffer (pH 7.5) could be described by a fractional conversion model for lower temperatures (55?57 °C) and a biphasic model for higher temperatures (58?70 °C). The enzyme showed a stable behavior toward high-pressure/temperature treatments. Keywords: Capsicum annuum; pepper; pectin methylesterase; purification; characterization; thermal and high-pressure stabilit

Pectic enzymes as potential enhancers of ascorbic acid production through the D-galacturonate pathway in Solanaceae

The increase of L-Ascorbic Acid (AsA) content in tomato (Solanum lycopersicum) is a common goal in breeding programs due to its beneficial effect on human health. To shed light into the regulation of fruit AsA content, we exploited a Solanum pennellii introgression line (IL12-4-SL) harbouring one quantitative trait locus that increases the content of total AsA in the fruit. Biochemical and transcriptomic analyses were carried out in fruits of IL12-4-SL in comparison with the cultivated line M82 at different stages of ripening. AsA content was studied in relation with pectin methylesterase (PME) activity and the degree of pectin methylesterification (DME). Our results indicated that the increase of AsA content in IL12-4-SL fruits was related with pectin de-methylesterification/degradation. Specific PME, polygalacturonase (PG) and UDP-D-glucuronic-acid-4-epimerase (UGlcAE) isoforms were proposed as components of the D-galacturonate pathway leading to AsA biosynthesis. The relationship between AsA content and PME activity was also exploited in PMEI tobacco plants expressing a specific PME inhibitor (PMEI). Here we report that tobacco PMEI plants, altered in PME activity and degree of pectin methylesterification, showed a reduction in low methylesterified pectic domains and exhibited a reduced AsA content. Overall, our results provide novel biochemical and genetic traits for increasing antioxidant content by marker-assisted selection in the Solanaceae family

Soluble Fermentable Dietary Fibre (Pectin) Decreases Caloric Intake, Adiposity and Lipidaemia in High-Fat Diet-Induced Obese Rats

Funding: This work was funded by the Scottish Government Rural and Environment Science and Analytical Services Division. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewedPublisher PD