Qi-Biao Wen

Functional properties and in vitro trypsin digestibility of red kidney bean (Phaseolus vulgaris L.) protein isolate: Effect of high-pressure treatment.

The effects of high-pressure (HP) treatment at 200-600MPa, prior to freeze-drying, on some functional properties and in vitro trypsin digestibility of vicilin-rich red kidney bean (Phaseolus vulgaris L.) protein isolate (KPI) were investigated. Surface hydrophobicity and free sulfhydryl (SH) and disulfide bond (SS) contents were also evaluated. HP treatment resulted in gradual unfolding of protein structure, as evidenced by gradual increases in fluorescence strength and SS formation from SH groups, and decrease in denaturation enthalpy change. The protein solubility of KPI was significantly improved at pressures of 400MPa or higher, possibly due to formation of soluble aggregate from insoluble precipitate. HP treatment at 200 and 400MPa significantly increased emulsifying activity index (EAI) and emulsion stability index (ESI); however, EAI was significantly decreased at 600MPa (relative to untreated KPI). The thermal stability of the vicilin component was not affected by HP treatment. Additionally, in vitro trypsin digestibility of KPI was decreased only at a pressure above 200MPa and for long incubation time (e.g., 120min). The data suggest that some physiochemical and functional properties of vicilin-rich kidney proteins can be improved by means of high-pressure treatment.

Formation of Amyloid Fibrils from Kidney Bean 7S Globulin (Phaseolin) at pH 2.0

The amyloid fibrils formed by heating 1.0% (w/v) kidney bean phaseolin (7S globulin) solution at pH 2.0 with an ionic strength of 20 mM at 85 degrees C were characterized using transmission electron microscopy (TEM), atomic force microscopy (AFM), binding of thioflavin T (Th T) and Congo Red dyes, and circular dichroism spectroscopy. The morphology of the formed fibrils was closely dependent upon heating time from 15 to 720 min. The diameters of the fibrils formed at various times were similar, but the mean contour length progressively increased with heating time. The Th T maximum fluorescence also progressively increased with heating time. The heating process caused remarkable changes in secondary, tertiary, and quaternary conformations of the phaseolin, but the extents of the changes were closely related to the heating time. With a short heating time (e.g., 15 min), the beta-strand content decreased from 38.7 to 22.9%, but further heating resulted in recovery of beta-strand structure. The tertiary and quaternary conformations gradually became flexible and unfolded upon heating. Gel electrophoresis analysis indicated that heating disrupted the polypeptides of phaseolin, leading to the formation of fragments with lower molecular mass (e.g., <10 kDa after 360 min). The results suggest that the amyloid fibril formation of phaseolin (7S globulin) involved the disruption of its polypeptides, as well as conformational changes at secondary, tertiary, and quaternary structural levels. This appears to be the first direct observation of amyloid fibrils from legume 7S storage globulin.

Functional and conformational properties of phaseolin (Phaseolus vulgris L.) and kidney bean protein isolate: a comparative study.

BACKGROUND Kidney bean (Phaseolus vulgris L.) seed is an underutilised plant protein source with good potential to be applied in the food industry. Phaseolin (also named G1 globulin) represents about 50 g kg(-1) of total storage protein in the seed. The aim of the present study was to characterise physicochemical, functional and conformational properties of phaseolin, and to compare these properties with those of kidney bean protein isolate (KPI). RESULTS Compared with kidney bean protein isolate (KPI), the acid-extracted phaseolin-rich protein product (PRP) had much lower protein recovery of 320 g kg(-1) (dry weight basis) but higher phaseolin purity (over 950 g kg(-1)). PRP contained much lower sulfhydryl (SH) and disulfide bond contents than KPI. Differential scanning calorimetry analyses showed that the phaseolin in PRP was less denatured than in KPI. Thermal analyses in the presence or absence of dithiothreitol, in combination with SH and SS content analyses showed the contributions of SS to the thermal stability of KPI. The analyses of near-UV circular dichroism and intrinsic fluorescence spectra indicated more compacted tertiary conformation of the proteins in PRP than in KPI. PRP exhibited much better protein solubility, emulsifying activity index, and gel-forming ability than KPI. The relatively poor functional properties of KPI may be associated with protein denaturation/unfolding, with subsequent protein aggregation. CONCLUSION The results presented here suggest the potential for acid-extracted PRP to be applied in food formulations, in view of its functional properties.

Wettability, surface microstructure and mechanical properties of films based on phosphorus oxychloride-treated zein.

BACKGROUND Zein, the predominant protein in corn, has been extensively studied as an alternative packaging material in edible and biodegradable films. However, films made from 100% zein are brittle under normal conditions. The aim of this investigation was to improve the film-forming properties of zein by chemical phosphorylation. The surface hydrophobicity, surface microstructure and mechanical properties of films based on untreated and phosphorus oxychloride (POCl(3))-treated zein were evaluated and compared. The effect of POCl(3) treatment on the rheological properties of zein solutions was also studied. RESULTS POCl(3) treatment, especially at pH 7 and 9, led to an increase in the apparent viscosity of zein solutions. Atomic force microscopy (AFM) analysis showed that the film based on POCl(3) -treated zein at pH 7 had a stone-like surface microstructure with a higher roughness (R(q)) than the untreated zein film. The AFM data may partially account for the phenomenon that this film exhibited high surface hydrophobicity (H(0) ). POCl(3) treatment diminished the tensile strength (TS) of zein films from 4.83-6.67 to 1.3-2.29 MPa. However, the elongation at break (EAB) of the films at pH 7 and 9 increased from 3.0-4.5% (control film) to 150.1-122.7% (POCl(3) -treated film), indicating the potential application of zein films in wrapping foods or in non-food industries such as sugar, fruit or troche that need good extension packing materials. CONCLUSION The data presented suggest that the properties of zein films could be modulated by chemical phosphorylation treatment with POCl(3) at an appropriate pH value.

Physicochemical and structural characterisation of protein isolate, globulin and albumin from soapnut seeds (Sapindus mukorossi Gaertn.)

The amino acid (AA) composition and physicochemical and conformational properties of protein isolate (SNPI), globulin (SNG) and albumin (SNA) fractions from soapnut seeds were evaluated. The essential AA of SNG, SNA and SNPI (except sulfur-containing AA) are sufficient for the FAO/WHO suggested requirements for 2-5year old infants. SNG and SNPI showed similar electrophoresis patterns and AA compositions, the subunit of those proteins consisted of two polypeptides linked by disulfide bonds. In contrast, SNA showed a different AA compositions and SDS-PAGE pattern. Both SNG and SNPI presented a typical U-shape protein solubility (PS)-pH profile, SNA showed a completely different PS-pH profile, especially at pH 2.0-4.0. The near-UV circular dichroism (CD), differential scanning calorimetry (DSC) and tryptophan fluorescence spectra analyses indicated that the flexibility in tertiary conformations decreased in the order: SNA>SNPI>SNG, while soapnut proteins had a similar secondary conformation, with a highly ordered structure (the β-types), as evidenced by far-UV CD spectra.

Conformational Study of Red Kidney Bean (Phaseolus vulgaris L.) Protein Isolate (KPI) by Tryptophan Fluorescence and Differential Scanning Calorimetry

Fluorescence and differential scanning calorimetry (DSC) were used to study changes in the conformation of red kidney bean (Phaseolus vulgaris L.) protein isolate (KPI) under various environmental conditions. The possible relationship between fluorescence data and DSC characteristics was also discussed. Tryptophan fluorescence and fluorescence quenching analyses indicated that the tryptophan residues in KPI, exhibiting multiple fluorophores with different accessibilities to acrylamide, are largely buried in the hydrophobic core of the protein matrix, with positively charged side chains close to at least some of the tryptophan residues. GdnHCl was more effective than urea and SDS in denaturing KPI. SDS and urea caused variable red shifts, 2-5 nm, in the emission λ(max), suggesting the conformational compactness of KPI. The result was further supported by DSC characteristics that a discernible endothermic peak was still detected up to 8 M urea or 30 mM SDS, also evidenced by the absence of any shift in emission maximum (λ(max)) at different pH conditions. Marked decreases in T(d) and enthalpy (ΔH) were observed at extreme alkaline and/or acidic pH, whereas the presence of NaCl resulted in higher T(d) and ΔH, along with greater cooperativity of the transition. Decreases in T(d) and ΔH were observed in the presence of protein perturbants, for example, SDS and urea, indicating partial denaturation and decrease in thermal stability. Dithiothreitol and N-ethylmaleimide have a slight effect on the thermal properties of KPI. Interestingly, a close linear relationship between the T(d) (or ΔH) and the λ(max) was observed for KPI in the presence of 0-6 M urea.

Conformational and thermal properties of phaseolin, the major storage protein of red kidney bean (Phaseolus vulgaris L.)

BACKGROUND A previous study of various functional and physicochemical properties of phaseolin indicated good potential of phaseolin for application in food formulations in view of its excellent functional properties. The aim of the present study was to explore the conformational and thermal properties of phaseolin in the presence of protein structural perturbants by intrinsic fluorescence emission spectroscopy and differential scanning calorimetry. Raman spectroscopy was also used to characterise the secondary structures of phaseolin. RESULTS The Raman spectrum of phaseolin indicated that β-sheets and random coils were the major secondary structures. Intrinsic fluorescence emission spectroscopy confirmed the structural peculiarity and compactness of phaseolin, as evidenced by the absence of any shift in emission maximum (λ(max)) in the presence of structural perturbants such as sodium dodecyl sulfate (SDS), guanidine hydrochloride, urea and dithiothreitol (DTT). Increasing NaCl concentration enhanced the thermal stability of phaseolin. Addition of chaotropic salts (1 mol L(-1)) caused progressive decreases in thermal stability following the lyotropic series of anions. Decreases in thermal denaturation temperature (T(d)) and enthalpy change (ΔH) were observed in the presence of protein perturbants such as SDS, urea and ethylene glycol, indicating partial denaturation and a decrease in thermal stability. DTT and N-ethylmaleimide had little effect on the thermal properties of phaseolin, confirming that phaseolin, a 7S globulin, is devoid of inter-polypeptide disulfide bonds. CONCLUSION The data presented here demonstrate the contributions of hydrophobic and electrostatic interactions and hydrogen bonding to the conformational stability of phaseolin.

Effects of acylation on the functional properties and in vitro trypsin digestibility of red kidney bean (Phaseolus vulgaris L.) protein isolate.

The effects of succinylation and acetylation on some functional properties and the in vitro trypsin digestibility of kidney bean protein isolate (KPI) were investigated. The extent of succinylation or acetylation progressively increased from 0% to 96% to 97%, as the anhydride-to-protein ratio increased from 0 to 1 g/g. Polyacrylamide gel electrophoresis (PAGE) and zeta potential analyses indicated that acylation, especially succinylation, considerably increased the net charge and hydrodynamic radius of the proteins in KPI, especially vicilin. Acylation treatment at various anhydride-to-protein ratios (0.05 to 1 g/g) remarkably improved the protein solubility (PS) and emulsifying activity index (EAI) at neutral pH, but the improvement by succinylation was much better than that by acetylation. Succinylation resulted in a marked decrease in mechanical moduli of heat-induced gels of KPI, while the mechanical moduli were, on the contrary, increased by acetylation. Additionally, in vitro trypsin digestibility was improved by the acylation in an anhydride-type and level-dependent manner. The results suggest that the functional properties of KPI could be modulated by the chemical acylation treatment, using succinic or acetic anhydride at appropriate anhydride-to-protein ratios.