Does harvesting age matter? Changes in structure and rheology of a shear-thickening polysaccharide from Cyathea medullaris as a function of age.

A shear-thickening polysaccharide from the New Zealand Black tree fern (Cyathea medullaris, commonly known as mamaku) extracted from different age fronds (stage 1: young, stage 2: fully grown and stage 3: old) was characterised in terms of structure and rheological properties. Constituent sugar analysis and 1H and 13C NMR revealed a repeating backbone of -4)-ß-D-GlcpA-(1 â†’ 2)-α-D-Manp-(1→, for all mamaku polysaccharide (MP) samples from different age fronds without any alterations in molecular structure. However, the molecular weight (Mw) was reduced with increasing age, from ~4.1 × 106 to ~2.1 × 106 Da from stage 1 to stage 3, respectively. This decrease in Mw (and size) consequently reduced the shear viscosity (ηs-Stage 1 > Î·s-Stage 2 > Î·s-Stage 3). However, the extent of shear-thickening and uniaxial extensional viscosity of MP stage 2 was greater than MP stage 1, which was attributed to a greater intermolecular interaction occurring in the former. Shear-thickening behaviour was not observed in MP stage 3.

The fate of mamaku gum in the gut: effect on in vitro gastrointestinal function and colon fermentation by human faecal microbiota.

Mamaku is a fern indigenous to the Pacific Islands with a long history of use for therapeutic benefits such as to combat skin conditions and manage gastrointestinal discomfort; however, the scientific understanding is limited. In this study, we examined the effect of mamaku gum, extracted from different age fronds of the New Zealand Black tree fern (Cyathea medullaris, Mamaku) (stage 1: young, stage 2: fully grown and stage 3: old), on gut function using in vitro models of static digestion, enzyme activity and static colonic fermentation. Under simulated gastric and small intestinal conditions, mamaku polysaccharide (MP) was indigestible as there was no decrease in the molecular weight (Mw) of the polymer. Mamaku gum could reduce the activity of digestive enzymes (α-amylase, pepsin and lipase) in a concentration-dependent manner, with the stage 1 sample showing the highest inhibition and stage 3 the lowest. All three mamaku gum samples could also equally bind bile acids during intestinal digestion. During fermentation, human faecal microbiota utilised the mamaku gum and significantly increased the production of total short-chain fatty acids (SCFAs) and reduced the pH when compared with the blank. However, changes in SCFAs and pH for mamaku groups were less prominent than for inulin and guar gum control groups, suggesting lower fermentability of mamaku gum compared to the latter two. Furthermore, mamaku gum altered the composition of colonic microbiota, specifically reducing the ratio of Firmicutes to Bacteroidetes and increasing the relative abundance of Bacteroides, Enterococcus, Paraprevotella and Parabacteroides genera. No obvious difference between mamaku gum samples from stage 1, 2 and 3 was observed during fermentation. Collectively, these results suggest that mamaku gum may modulate the functionality of the host gut by reducing enzyme activity, binding bile acids, altering the colonic microbial composition and producing SCFAs.

Complexation of Anthocyanin-Bound Blackcurrant Pectin and Whey Protein: Effect of pH and Heat Treatment.

A complexation study between blackcurrant pectin (BCP) and whey protein (WP) was carried out to investigate the impact of bound anthocyanins on pectin−protein interactions. The effects of pH (3.5 and 4.5), heating (85 °C, 15 min), and heating sequence (mixed-heated or heated-mixed) were studied. The pH influenced the color, turbidity, particle size, and zeta-potential of the mixtures, but its impact was mainly significant when heating was introduced. Heating increased the amount of BCP in the complexes­especially at pH 3.5, where 88% w/w of the initial pectin was found in the sedimented (insoluble) fraction. Based on phase-separation measurements, the mixed-heated system at pH 4.5 displayed greater stability than at pH 3.5. Heating sequence was essential in preventing destabilization of the systems; mixing of components before heating produced a more stable system with small complexes (<300 nm) and relatively low polydispersity. However, heating WP before mixing with BCP prompted protein aggregation­producing large complexes (>400 nm) and worsening the destabilization. Peak shifts and emergence (800−1200 cm−1) in infrared spectra confirmed that BCP and WP functional groups were altered after mixing and heating via electrostatic, hydrophobic, and hydrogen bonding interactions. This study demonstrated that appropriate processing conditions can positively impact anthocyanin-bound pectin−protein interactions.

Rheology, Microstructure, and Storage Stability of Emulsion-Filled Gels Stabilized Solely by Maize Starch Modified with Octenyl Succinylation and Pregelatinization.

We prepared emulsion-filled gels stabilized using octenyl succinic anhydride-modified and pregelatinized maize starch (OSA-PGS). The effect of the oil volume fraction (Φ, 0.05-0.20) and OSA-PGS concentration (3-10% w/v) on the rheological and microstructural properties of the emulsion-filled gels was evaluated. Confocal fluorescence images showed that OSA-PGS stabilized the emulsion, indicated by the formation of a thick layer surrounding the oil droplets, and simultaneously gelled the aqueous phase. All of the emulsions exhibited shear-thinning flow behavior, but only those with 10% w/v OSA-PGS were categorized as Herschel-Bulkley fluids. The rheological behavior of the emulsion-filled gels was significantly affected by both the OSA-PGS concentration and Φ. The mean diameters (D1,0, D3,2, and D4,3) of oil droplets with 10% w/v OSA-PGS were stable during 30 days of storage under ambient conditions, indicating good stability. These results provide a basis for the design of systems with potential applications within the food industry.

Molecular, rheological and physicochemical characterisation of puka gum, an arabinogalactan-protein extracted from the Meryta sinclairii tree.

A water-soluble polysaccharide (type II arabinogalactan-protein) extracted from the gum exudate of the native New Zealand puka tree (Meryta sinclairii), was characterised for its molecular, rheological and physicochemical properties. In 0.1 M NaCl, the weight average molecular weight (Mw) of puka gum is 5.9 × 106 Da with an RMS radius of 56 nm and z-average hydrodynamic radius of 79 nm. The intrinsic viscosity of the polysaccharide is 57 ml/g with a coil overlap concentration 15% w/w. Together, the shape factor, p, of 0.70 (exponent of RMS radius vs. hydrodynamic radius), Smidsrød-Haug's stiffness parameter B of 0.031 and Mark-Houwink exponent α of 0.375 indicate that the polysaccharide adopts a spherical conformation in solution, similar to gum arabic. The pKa is 1.8. The polysaccharide exhibits a Newtonian to shear-thinning behaviour from 0.2 to 25% w/w. Viscosity of the polysaccharide (1 s-1) decreases with decreasing concentration, increasing temperature, ionic strength, and at acidic pH.

Kinetic stability and cellular uptake of lutein in WPI-stabilised nanoemulsions and emulsions prepared by emulsification and solvent evaporation method.

The particle size and lutein encapsulation efficiency of nanoemulsions prepared by emulsification and solvent evaporation method were 68.8±0.3nm and 80.7±0.8%, respectively, whereas they were 147.3±0.6nm and 86.3±0.3% for conventional emulsions. All the emulsions had no change in their particle size during storage (28days at 5, 20 and 40°C) but their lutein content and emulsion colour decreased, especially at 40°C. The lutein emulsions were analysed using MTT assay on the gut enterocyte cell line Caco-2 and they showed no toxicity as the cell viability was more than 80% at 10times or higher dilution after 24h of incubation. However, there was a higher cellular uptake of lutein by Caco-2 cells in nanoemulsions (872.9±88.3pmol/mgprotein) than conventional emulsions (329.5±214.6pmol/mgprotein). The results of this study indicated that nanoemulsions can be used as a delivery system to improve the cellular uptake of lutein.

Physicochemical properties of whey protein, lactoferrin and Tween 20 stabilised nanoemulsions: Effect of temperature, pH and salt.

Oil-in-water nanoemulsions were prepared by emulsification and solvent evaporation using whey protein isolate (WPI), lactoferrin and Tween 20 as emulsifiers. Protein-stabilised nanoemulsions showed a decrease in particle size with increasing protein concentration from 0.25% to 1% (w/w) level with Z-average diameter between 70 and 90 nm. However, larger droplets were produced by Tween 20 (120-450 nm) especially at concentration above 0.75% (w/w). The stability of nanoemulsions to temperature (30-90°C), pH (2-10) and ionic strength (0-500 mM NaCl or 0-90 mM CaCl2) was also tested. Tween 20 nanoemulsions were unstable to heat treatment at 90°C for 15 min. WPI-stabilised nanoemulsions exhibited droplet aggregation near the isoelectric point at pH 4.5 and 5 and they were also unstable at salt concentration above 30 mM CaCl2. These results indicated that stable nanoemulsions can be prepared by careful selection of emulsifiers.

The cation-controlled and hydrogen bond-mediated shear-thickening behaviour of a tree-fern isolated polysaccharide.

The shear-thickening rheological behaviour (between 5 and 20s(-1)) of a 5% (w/w) viscoelastic gum extracted from the fronds of the native New Zealand black tree fern or mamaku in Maori was further explored by manipulating the salt content. The freeze-dried mamaku gum contained a high mineral content and sugars which upon removal via dialysis, resulted in the loss of shear thickening. However, this loss was reversible by the addition of salts to the dialysed dispersion. The mechanism of shear-thickening behaviour was therefore hypothesised to be due to shear-induced transition of intra- to intermolecular hydrogen bonding, promoted by the screening effect of cations. Mono-, di- and trivalent salts, i.e. Na(+), K(+), N(CH3)4(+), Ca(2+), Mg(2+), Al(3+) and La(3+) at concentrations between 0.001 and 1.0M were tested to support the hypothesis as well as to demonstrate the sensitivity of the biopolymer to cation valency and concentrations. The cation valency and concentration were crucial factors in determining: (i) zero-shear viscosity, (ii) critical shear rate, γ˙c (or shear rate at the onset of shear-thickening) and (iii) the extent of shear-thickening of the solution. For mono- and divalent cations these parameters were similar at equivalent ionic strengths and fairly independent of the cation type. Trivalent cations (La(3+)) however caused precipitation of the gum in the concentration range of 0.005-0.05 M but clear dispersions were obtained above 0.05 M.

Extraction and characterisation of pomace pectin from gold kiwifruit (Actinidia chinensis).

Gold kiwifruit pomace extracted using citric acid, water and enzyme (Celluclast 1.5L) were studied in terms of pectin yield, protein, ash, non-starch polysaccharide, galacturonic acid (GalA), neutral sugar composition, molar mass (Mw), viscosity and degree of branching. Water-extracted pectin was considered closest to its native form. Enzyme extracted pectin showed the highest yield (∼ 4.5%w/w) as compared with the acid and water extraction methods (∼ 3.6-3.8%w/w). Pectin obtained from different extraction methods showed different degree of branching. The Mw and root mean square (RMS) radius varied with the extraction methods with values of 8.4 × 10(5) g/mol and 92 nm, 8.5 × 10(5)g/mol and 102 nm, 6.7 × 10(5) g/mol and 52 nm for acid, water and enzymatic extraction methods, respectively. Similar trend was observed for pectin viscosity, with water-extracted pectin giving a slightly higher viscosity followed by acid and enzyme-extracted pectin. This study showed that gold kiwifruit pomace pectin has potential application in food products.

Probing hydrogen bond interactions in a shear thickening polysaccharide using nonlinear shear and extensional rheology.

Mamaku gum is a polysaccharide extracted from the fronds of the black tree fern found in New Zealand. The cooked pith has traditionally been used for various medicinal purposes and as a food source by the Maori people of New Zealand. It has potential applications as a thickener in the food industry and as a palliative for patients with dysphagia. Studies on the shear rheology of Mamaku gum have revealed that the gum exhibits shear thickening at a critical shear rate due to a transition from intra- to inter-molecular chain interactions upon shear-induced chain elongation. In this paper, we demonstrate that these interactions are primarily due to hydrogen bonding. We perform extensional rheology on mixtures of Mamaku gum and urea (a known disruptor of hydrogen bonds) to quantify the nature of these interactions. Capillary Breakup Extensional Rheometry (CaBER) performed on the pure Mamaku gum solutions yield plateau values of the Trouton ratio as high as ∼10(4), showing that the viscoelasticity of the gum in uniaxial elongation is much higher than in shear. For all Mamaku concentrations tested, the extensional viscosity decreases upon increasing urea concentration. Furthermore, the relaxation time decreases exponentially with increasing urea concentration. This exponential relationship is independent of the Mamaku concentration, and is identical to the relationships between urea concentration and characteristic timescales measured in nonlinear shear rheology. We show using the sticky reptation model for polymers with multiple sticker groups along the backbone how such a relationship is consistent with a linear decrease in the free energy for hydrogen bond dissociation. We then demonstrate that a time-concentration superposition principle can be used to collapse the viscoelastic properties of the Mamaku-gum/urea mixtures.

Characterization of gold kiwifruit pectin from fruit of different maturities and extraction methods.

Studies on gold kiwifruit pectins are limited. In this work, the characterization of pectin isolated from two different stages of maturity of gold kiwifruit, namely early harvested fruit (EHF) and main harvested fruit (MHF) isolated by three methods (acid, water, enzymatic) was carried out. Pectins isolated from MHF were higher in galacturonic acid content (52-59% w/w) and weight-average molecular weights (Mw, 1.7-3.8 × 10(6)g/mol) compared with EHF pectins (29-49% w/w and 0.2-1.7 × 10(6)g/mol respectively). Enzymatic treatment gave the highest yield but lowest in Mw, viscosity and mechanical spectra for both maturities. The pectin of both maturities was classified as high-methoxyl pectin with the degree of esterification ranged from 82% to 90%. Water-extracted MHF pectin molecules had the highest RMS radius (182.7 nm) and Mw (3.75 × 10(6)g/mol). The water extraction method appeared to retain the native state of pectin molecules compared with acid and enzymatic extraction methods based on the Mw and viscosity data.

Structure of a shear-thickening polysaccharide extracted from the New Zealand black tree fern, Cyathea medullaris.

A shear-thickening water-soluble polysaccharide was purified from mucilage extracted from the fronds of the New Zealand black tree fern (Cyathea medullaris or 'mamaku' in Maori) and its structure characterised. Constituent sugar analysis by three complementary methods, combined with linkage analysis (of carboxyl reduced samples) and 1H and 13C nuclear magnetic resonance spectroscopy (NMR) revealed a glucuronomannan comprising a backbone of 4-linked methylesterified glucopyranosyl uronic acid and 2-linked mannopyranosyl residues, branched at O-3 of 45% and at both O-3 and O-4 of 53% of the mannopyranosyl residues with side chains likely comprising terminal xylopyranosyl, terminal galactopyranosyl, non-methylesterified terminal glucopyranosyl uronic acid and 3-linked glucopyranosyl uronic acid residues. The weight-average molecular weight of the purified polysaccharide was ∼1.9×10(6) Da as determined by size-exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALLS). The distinctive rheological properties of this polysaccharide are discussed in relation to its structure.

Effect of Celluclast 1.5L on the physicochemical characterization of gold kiwifruit pectin.

The effects of Celluclast 1.5L concentration on the physicochemical characterization of gold kiwifruit pectin was evaluated. Varying the enzyme concentration affected the pectin yield and pectin physicochemical properties. The viscosity of extracted pectin was largely dependent on the enzyme concentration. Celluclast 1.5L with medium concentration exhibited the highest viscosity. Varying the enzyme concentration also influenced the molecular weight distribution. High molecular weight (M(w)) pectin (1.65 × 10(6) g/mol) was obtained when the medium concentration was used. Overall, the study clearly reflects the importance of taking into consideration the amount of cellulytic enzyme added in order to determine the final quality of pectin.

Complex rheological properties of a water-soluble extract from the fronds of the black tree fern, Cyathea medullaris.

A water-soluble extract was obtained from the fronds of a New Zealand native black tree fern (Cyathea medullaris or Mamaku in Maori). The extract exhibited complex rheological behavior. Newtonian, shear-thinning, shear-thickening, thixotropic, antithixotropic, and viscoelastic behaviors were observed depending on polymer concentration, shear rate, and shear history. The extract also displayed rod-climbing and self-siphoning properties typical of viscoelastic fluids. Such complex rheological properties have been reported in synthetic or chemically modified polymers but are less frequent in unmodified biopolymers. Although Mamaku extract obtained from the pith of the fern has been traditionally used by the Maori in New Zealand for treating wounds and diarrhea among other ailments, this material has never been characterized before. This study reports on the chemical composition of the extract and on its viscoelastic properties through rotational and oscillatory rheological measurements. Explanations of the mechanism behind the rheological properties were based on transient network models for associating polymers.

Rheological and light scattering properties of flaxseed polysaccharide aqueous solutions.

Polysaccharides isolated from flaxseed meals using ethanol consisted of a soluble ( approximately 7.5% w/w) and an insoluble fraction (2% w/w). The soluble fraction was dialyzed in various salt concentrations and characterized using viscometry and light scattering techniques. Observations using a size-exclusion column coupled to a multiangle laser light scattering (SEC-MALLS) revealed three molecular weight fractions consisting of a small amount ( approximately 17%) of large molecular weight species (1.0 x 10(6)) and a large amount ( approximately 69%) of small molecular weight species (3.1 x 10(5) Da). Dynamic light scattering measurements indicated the presence of very small molecules (hydrodynamic radius approximately 10 nm) and a very large molecular species (hydrodynamic radius in excess of 100 nm); the latter were probably aggregates. The intrinsic viscosity, [eta], of the polysaccharide in Milli-Q water was 1030 +/- 20 mL/g. The viscosity was due largely to the large molecular weight species since viscosity is influenced by the hydrodynamic volume of molecules in solution. The Smidsrod parameter B obtained was approximately 0.018, indicating that the molecules adopted a semi-flexible conformation. This was also indicated by the slope ( approximately 0.56) from the plot of root-mean-square (RMS) radius versus molar mass (M(w)).

Development of an improved procedure for isolation and purification of exopolysaccharides produced by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2483.

A method was developed for the isolation and purification of exopolysaccharide (EPS) produced by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2483 that can be adapted for industrial-scale operation. Hydrolyzed milk medium, which was ultrafiltered to remove molecular species larger than 2.5 x 10(5) Da, was found to be a suitable growth medium for the bacteria, which produced approximately 400 mg EPS/l . Optimal isolation of EPS was achieved using centrifugation, filtration and ethanol precipitation methods. Insoluble and soluble EPS fractions were obtained. The soluble fraction was purified using a series of ethanol precipitations to achieve approximately 98% (w/w) purity. This fraction consisted of galactose, glucose, rhamnose and mannose in the ratio of approximately 5:1:0.6:0.5, with traces of glucosamine.