Influence of inulin/oligofructose on the acid‐induced cold aggregation and gelation of preheated soy proteins

BACKGROUND: In recent years inulin‐type prebiotics have attracted much attention due to consumers' awareness of the health benefits of functional foods. Currently no information is available about the possible texture‐modifying effect of these non‐ionizable polar carbohydrates in different soy‐based food systems. In this study, the effect of inulin/oligofructose on the cold aggregation and gelation of preheated soy protein isolate (SPI) and its fractions (7S, 11S, and their mixture), induced by glucono‐δ‐lactone (GDL), were evaluated by turbidity (A₆₀₀) and dynamic rheological measurements. RESULTS: Oligofructose significantly delayed the aggregation of all protein samples and decreased the end‐point optical density of 11S fraction and SPI. Inulin, a long‐chain fructan, only delayed the aggregation of 7S globulin and reduced the capacity of aggregation (A₆₀₀) of SPI. While oligofructose showed no significant effect, the addition of 5% (w/v) inulin enhanced the gelation of SPI and the 7S/11S mixture, which was demonstrated by the increase in gel storage modulus up to 13.6% and 10.1% (P < 0.05), respectively. CONCLUSION: Inulin was found to enhance the viscoelastic properties of GDL‐induced cold‐set soy protein gels. It is expected that ‘functional’ cold‐set gel products with improved texture can be prepared from preheated soy proteins and inulin. Copyright © 2009 Society of Chemical Industry     

Effects of inulin and oligofructose on the rheological characteristics and probiotic culture survival in low-fat probiotic ice cream

ABSTRACT:  The effects of supplementation of oligofructose or inulin on the rheological characteristics and survival of Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 in low-fat ice cream stored at –18 °C for 90 d were studied. Addition of oligofructose or inulin to ice cream mix significantly increased apparent viscosity and overrun and developed the melting properties in ice cream during storage ( P < 0.05). However, the highest increase in firmness, the lowest change in melting properties, and the longest 1st dripping time were obtained in probiotic ice cream containing inulin ( P < 0.05). Some textural properties have also improved especially by the end of storage. Freezing process caused a significant decrease in the viability of Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 ( P < 0.05). Oligofructose significantly improved the viability of L. acidophilus La-5 and B. animalis Bb-12 in ice cream mix ( P < 0.05). Although the viable numbers for both bacteria decreased throughout the storage, the minimum level of 10⁶ CFU/g was maintained for B. animalis Bb-12 in only ice cream with oligofructose during storage.     

Antioxidant Activity of Soy Protein Hydrolysates in a Liposomal System

ABSTRACT: Native and heated soy protein isolate was hydrolyzed with 3 purified (pepsin, papain, and chymotrypsin) and 3 crude (Alcalase®, Protamex^TM, and Flavourzyme^TM) proteases. The hydrolysates were incubated (37 °C, 1 h) with a liposome-oxidizing system (50 μM FeCl₃/0.1 mM ascorbate, pH 7.0) to test antioxidant activities by determining the concentrations of TBARS. Degree of hydrolysis of SPI hydrolysates ranged from 1.7 to 20.6%. Both hydrolyzed and nonhydrolyzed SPI decreased TBARS (by 28 to 65%), except for papain-hydrolyzed samples. Samples of chymotrypsin- and Flavourzyme-hydrolyzed (0.5 h) preheated SPI had the greatest inhibitory effect on lipid oxidation.     

Alginate- and gellan-based edible films for probiotic coatings on fresh-cut fruits

ABSTRACT:  Alginate- (2% w/v) or gellan-based (0.5%) edible films, containing glycerol (0.6% to 2.0%), N-acetylcysteine (1%), and/or ascorbic acid (1%) and citric acid (1%), were formulated and used to coat fresh-cut apple and papaya cylinders. Water vapor permeability (WVP) was significantly higher ( P < 0.05) in alginate films (0.30 to 0.31 × 10⁻⁹g m/Pa s m²) than in the gellan ones (0.26 to 0.27 × 10⁻⁹g m/Pa s m²). Addition of 0.025% (w/v) sunflower oil decreased WVP of gellan films (0.20 to 0.22 × 10⁻⁹ g m/Pa s m²). Water solubility of gellan and alginate films at 25 °C (0.47 to 0.59 and 0.74 to 0.79, respectively) and their swelling ratios (2.3 to 2.6 and 1.6 to 2.0, respectively) indicate their potential for coating high moisture fresh-cut fruits. Fresh-cut apple and papaya cylinders were successfully coated with 2% (w/v) alginate or gellan film-forming solutions containing viable bifidobacteria. WVP in alginate (6.31 and 5.52 × 10⁻⁹g m/Pa s m²) or gellan (3.65 and 4.89 × 10⁻⁹ g m/Pa s m²) probiotic coatings of papaya and apple, respectively, were higher than in the corresponding cast films. The gellan coatings and films exhibited better water vapor properties in comparison with the alginate coatings. Values > 10⁶ CFU/g B. lactis Bb -12 were maintained for 10 d during refrigerated storage of fresh-cut fruits, demonstrating the feasibility of alginate- and gellan-based edible coatings to carry and support viable probiotics on fresh-cut fruit.     

Interaction of Myofibrillar and Preheated Soy Proteins

ABSTRACT: Soy protein isolate (SPI) was preheated at 90 °C and 95 °C for 3 min to obtain preheated samples, SPI₉₀ and SPI₉₅, respectively. The preheat treatment increased protein hydrophobicity and decreased the aggregation of 11S acidic and basic subunits. The 7S and 11S soy proteins exhibited a decreased thermal stability when mixed with pork myofibrillar protein isolate (MPI). The presence of preheated SPI accelerated the disappearance of myosin heavy chain in the gelling process. Incorporation of preheated SPI significantly increased the MPI gel elasticity and hardness while native SPI showed negative effects.     

BOUND WATER IN FRUIT PRODUCTS BY THE FREEZING METHOD

SUMMARY— An insulated heat-sink containing Freon-12® (N.B.P. −21.6°F) provides a reproducible system for measurement of thermal properties of fruit products. The difference in time required to remove the latent heat of fusion of the "eutectic" mixture in comparison with distilled water measures the "bound" water. The 34-37%"bound" water in papaya (var. solo) pulp is unaffected by varying pH in the range 3.0–6.0. Less than 10% water is bound in guava, passion fruit and pineapple juice products with up to 35% sucrose added. Thermal conductivity of the solid phase "eutectic" mixture in the 5–50% soluble solids range fits the regression Y = AB^x, Y = (3.69) (0.96)^x where x = percent total soluble solids (TSS) and Y = thermal conductivity (Cal x 10³/(cm²) (sec) (°C/cm)). The relationship of freezing point to TSS values as 1.10 + 0.64 (TSS) in the test system is significant (P < 0.01) in the range 5–50% TSS. The time for fusion (θ_w) of sucrose/water in the range 5–50% TSS fits the regression: log θ_w= (−0.8325) (−0.0165) (TSS) or θ_w= (0.984) (0.164)^TSS sec. Addition of solutes such as sucrose will inhibit gelation without affecting "bound" water values. Pectin gel structures apparently are dependent on secondary bonding and independent of "bound" water per se.     

Elucidation of structural changes in soy protein isolate upon heating by Raman spectroscopy

Changes in protein secondary and tertiary structure of soy protein isolate (SPI) upon heating process were investigated by Raman spectroscopy. SPI refrigerated and heated at 70, 100 and 200 °C was analysed. Non-significant differences in secondary structure (α-helix, β-sheet, unordered and turn) were observed in SPI upon heating at these temperatures. However, comparison of the Raman spectra of SPI refrigerated and heated at 70, 100 and 200 °C reveals changes in hydrophobic group environments. An intensity decreasing trend of the 1340 cm⁻¹ band attributable to tryptophan vibrations was observed, which involves an increase in solvent exposure of tryptophan residues. The spectral results also showed intensity increasing trends for the 1450 and 2935 cm⁻¹ bands of SPI upon heating, which can be ascribed to breaking of hydrophobic contacts and subsequent solvent exposure of the corresponding hydrophobic aliphatic groups. Finally, it was found tertiary structural transitions towards greater proportions of buried tyrosine residues upon heating of SPI.     

Production of beta-glucosidase and hydrolysis of isoflavone phytoestrogens by Lactobacillus acidophilus, Bifidobacterium lactis, and Lactobacillus casei in soymilk

ABSTRACT:  The study determined β-glucosidase activity of commercial probiotic organisms for hydrolysis of isoflavone to aglycones in fermenting soymilk. Soymilk made with soy protein isolate (SPI) was fermented with Lactobacillus acidophilus LAFTI^® L10, Bifidobacterium lactis LAFTI^® B94, and Lactobacillus casei LAFTI^® L26 at 37 °C for 48 h and the fermented soymilk was stored for 28 d at 4 °C. β-Glucosidase activity of organisms was determined using ρ-nitrophenyl β-D-glucopyranoside as a substrate and the hydrolysis of isoflavone glycosides to aglycones by these organisms was carried out. The highest level of growth occurred at 12 h for L. casei L26, 24 h for B. lactis B94, and 36 h for L. acidophilus L10 during fermentation in soymilk. Survival after storage at 4 °C for 28 d was 20%, 15%, and 11% greater ( P < 0.05) than initial cell counts, respectively. All the bacteria produced β-glucosidase, which hydrolyzed isoflavone β-glycosides to isoflavone aglycones. The decrease in the concentration of β-glycosides and the increase in the concentration of aglycones were significant ( P < 0.05) in the fermented soymilk. Increased isoflavone aglycone content in fermented soymilk is likely to improve the biological functionality of soymilk.     

Factors controlling the growth of Clostridium botulinum types A and B in pasteurized, cured meats III. The effect of potassium sorbate

The growth of Clostridium botulinum types A and B spores, at 10¹ or 10³ per container, was studied in a pork slurry system containing nitrite (40 μg/g), sodium chloride (2.5, 3.5, 4.5% w/v) sodium isoascorbate (550 μg/g) at varying pH levels, with or without potassium sorbate (0.26% w/v), without heating and after two heat treatments (80°C for 7 min, and 80°C for 7 min + 70°C for 1 hr) followed by storage at 15, 17.5, 20 or 35°C for up to 6 months. At a given spore inoculum, potassium sorbate significantly decreased toxin production, as did increasing NaCl, decreasing pH or decreasing storage temperature. Heat treatment did not significantly affect spoilage or toxin production overall, but interacted significantly with some factors. The effect of sorbate was greater at 3.5% NaCl than at 2.5%, at pH values below 6.0, and at low storage temperature.     

UNIAXIAL COMPRESSION OF GELS MADE FROM PROTEIN AND K-CARRAGEENAN

Investigations of gels (18% total solids) made from pea protein isolates (PPI) or soy protein isolate (SPI) with differing amounts of _K-carrageenan added showed that the gel strength increased with the concentration of _K-carrageenan. When the concentration of_K-carrageenan exceeded 0.4%, gels made with PPI were stronger and more stiff than equivalent gels made with SPI. Addition of _K-carrageenan stabilized gels made with PPI towards variations in brittleness (indicated by strain at fracture) with pH. This was not the case when SPI was used. Preheating (75C, 2 min) suspensions containing protein isolates and _K-carrageenan before gel formation increased the strength and stiffness of the final gels, most pronounced when SPI was used. Addition of NaCl (0.5–2%) reduced strength and stiffness of gels, whereas CaCl₂ had no influence on gel properties. Mixtures of PPI and SPI proved to be weaker and more brittle than gels made from only SPI of PPI. Results indicate that using proteins of different origin can cause differences in gel structure.     

Purification and Characterization of an α-L-Rhamnosidase from Aspergillus terreus of Interest in Winemaking

ABSTRACT: An enzyme with α-L-rhamnosidase activity was purified to homogeneity from a culture filtrate of Aspergillus terreus after growth in a medium containing L-rhamnose as the sole carbon source. The biosynthesis of this enzyme was repressed by glucose. The enzyme had a molecular mass of 96 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an isoelectric point of 4.6 as determined by analytical isoelectric focusing. The pH and temperature optima for the enzyme were found to be 4.0 and 44 °C, respectively. Using p-nitrophenyl-α-L-rhamnopyranoside as a substrate, the enzyme exhibited Michaelis-Menten kinetics with K_M and V_max values of 0.17 mM and 84 U/mg, respectively. The enzyme was inhibited competitively by L-rhamnose (K₁ 2.5 mM). Divalent cations such as Ca²⁺ Mg²⁺ Zn²⁺ and Co²⁺ stimulated the a-L-rhamnosidase activity, whereas this was inhibited by Hg2+ and Cd2+. Ethanol (12% v/v) and glucose (21% w/v) decreased enzyme activity by approximately 20%, while this was not affected by SO₂.     

Autolysis and Protease Inhibition Effects on Dynamic Viscoelastic Properties during Thermal Gelation of Squid Muscle

ABSTRACT: Viscoelastic changes during thermal gelation of squid ( Loligo vulgaris ) muscle with protease inhibitors were studied, in order to evaluate the contribution of different proteinases to gel degradation. Calcium chloride was also tested as enzyme activator. A minimum in elastic modulus without inhibitors was achieved at 38 to 40°C. Although denaturation temperature was around 55°C, pronounced melting of connective tissue started at 40°C. Proteolysis occurred between 25 and 75°C, with maximum at 40 to 45°C. Addition of PMSF led to highest values of G'. The strong inhibition of autolytic activity by PMSF confirmed the predominance of serine proteases. Addition of CaCl₂ favored thermal protein aggregation from 40°C upwards.     

Molecular interactions and functionality of a cold-gelling soy protein isolate

ABSTRACT:  A soy protein isolate (SPI) was thermally denatured at a critical concentration of 8% protein for 3 h at 95 °C, resulting in a powder that was readily reconstituted at ambient temperature and that demonstrated improved heat stability and cold-set gel functionality when compared to a control SPI. When SPI was heated at 3% protein equivalently, prior to reconstitution to 8% protein, the final viscosity was about 3 orders of magnitude less than the original sample. The viscosity of SPI heated at 3% protein was still nearly 2 orders of magnitude less than the original sample after both samples were reheated at 8% protein. These results suggested that heat denaturation at low protein concentrations limited network formation even after the protein concentration and interaction sites increased, impacting the isolate's cold gelling ability. Gelation was prevented upon treatment of SPI with iodoacetamide, which carbaminomethylated the cysteine residues, establishing the role of disulfide bonds in network formation. The viscosity of the 8% protein dispersion was also reduced by 2 orders of magnitude when treated with 8 M urea, and when combined with 10 mM DTT the gel viscosity was decreased by another order of magnitude. These results suggested that hydrophobic interactions played a primary role in gel strength after disulfide bonds form. The need for a higher concentration of protein during the heating step indicated that the critical disulfide bonds are intermolecular. Ultimately, the functionality produced by these protein–protein interactions produced a powdered soy protein isolate ingredient with consistent cold-set and thermal gelation properties.     

Thermorheological Characteristics of Soybean Protein Isolate

ABSTRACT: Small amplitude oscillation shear measurement was used to study gel rigidity of commercial soy protein isolate (SPI) dispersions during isothermal and non-isothermal heating. Temperature sweep data (20°C to 90°C atheating rate of 1°C/min) of SPI dispersions demonstrated that elastic modulus (G) predominates over viscous component (G') for all concentrations studied. The gelation kinetics of SPI was evaluated by a non-isothermal technique as a function of elastic modulus (G). During experiments, it was observed that a critical concentration of 10% was required to form a true SPI gel. Thermorheological data of 10% and 15% SPI dispersions were adequately fitted by 2nd-order reaction kinetics. The reaction order of gelation was initially calculated by multiple regression technique correlating dG'/dt, G'and temperature, which finally was verified by linear regression of kinetic equation at selected order. Isothermal data of 15% SPI was also followed by 2nd-order reaction kinetics. The activation energy during the isothermal technique was significantly higher than non-isothermal gelation of SPI at same concentration level. Gel strength of the non-isothermally heated SPI sample (15% to 20%) was compared with isothermally heated (90°C for 30 min) one. Higher protein concentration (20%) and isothermal heating exhibited significantly higher gel rigidity while the difference between the 2 processes was insignificant at 15% concentration at a similar condition.     

Time-temperature studies of gellan polysaccharide-high sugar mixtures: effect of sodium ions on structure formation

ABSTRACT:  Changes to the viscoelastic storage and loss moduli were measured as a function of temperature and oscillatory frequency for 0.5% (w/w) gellan:80% (w/w) cosolute dispersions with added Na⁺ (40 to 160 mM). Isothermal frequency (0.15 to 15 Hz) and thermal scans (at 0.15 Hz) were performed over a decreasing then increasing temperature range of 85 to 5 °C and 5 to 85 °C, respectively. Moduli were found to increase in magnitude with decreasing temperature and increasing levels of Na⁺ during cooling, then remained relatively thermally irreversible upon heating. Isothermal frequency (ITF) data were described using the time–temperature superposition (TTS) principle and the modified Cole–Cole (MCC) analysis. Both TTS and the MCC analyses successfully described the behavior of samples containing 40 mM added Na⁺ during cooling and heating, and at the 100-mM Na⁺ level during cooling. TTS superposed ITF data over the entire temperature range, whereas successful superposition was restricted to lower temperatures in the MCC analysis, where the viscoelastic response was dominated by the long-range relaxation of gellan chains between junction zones. Failure of both analyses was attributed to the formation of junction zones composed of polymer–polymer associations. It is proposed that the addition of Na⁺ promotes the formation of a weakly cross-linked gellan network.     

Isolation of Tomato Seed Meal Proteins with Salt Solutions

Salt solutions were used in isolating tomato seed meal proteins. Na₂SO₃ and NaCl solutions at different concentrations, and pH were included in a central composite design to find optimum conditions of protein isolation. The highest total protein yield was achieved with water extraction (no salt present). Salt extraction at pH 7.5 produced isolates with protein content of 93.4% (NaCl 5% w/v) and 77.1% (Na₂SO₃ 0.5% w/v). Observed values were in good agreement with predicted values. Isolates extracted with different salt solutions ranged from less soluble but very resistant to heat and Ca²⁺, to very surface active with functional properties comparable to commercial soy isolates.     

Effect of inulin on the rheological properties of silken tofu coagulated with glucono-δ-lactone

This study investigated the rheological properties of inulin-containing silken tofu coagulated with glucono-δ-lactone (GDL) upon heating. Inulin (Raftiline^® HP-gel) was added to a soy protein isolate-enriched cooked soymilk at 0%, 1%, 2%, 3% and 4% (w/v) levels along with 0.4% (w/v) GDL to prepare acid-induced silken tofu. Gelation was induced by heating the soymilk mixture from 20 to 90 °C at a constant rate (1 °C/min) or isothermally at 90 °C for 30 min. The gelling properties were measured with dynamic small-deformation mechanical analysis and static large-deformation compression tests. The rheological changes in soymilk during gelation were dependent upon both the pH decline (hydrolysis of GDL) and the specific temperature of heating. Control samples heated to 50 °C, with the pH lowered to 5.95, started to gel, showing a rapid increase in storage (G′) and loss (G″) moduli afterwards. The addition of 2% inulin lowered the on-set gelling temperature by 2.8 °C and improved (P < 0.05) both rheological parameters of the tofu gel as well as hardness and rupture force (textural profile analysis) of the formed silken tofu. The results indicated that inulin enhances the viscoelastic properties of GDL-coagulated silken tofu, and the textural effect of inulin is an added benefit to its current application mainly as a prebiotical ingredient in food.     

Whey Protein Film Composition Effects on Potassium Sorbate and Natamycin Diffusion

ABSTRACT: To assess the ability of whey protein films to act as antimicrobial carriers, the effect of film composition on preservative diffusion was investigated. Preservative diffusion coefficients were measured at 24°C in whey protein isolate (WPI) films with different WPI-glycerol plasticizer ratios (1:1 to 15:1), beeswax (BW) content, 0% to 40% w/w dry solids, and preservative addition of 0.3% (w/w) natamycin or 1.6% (w/w dry solids) potassium sorbate. Diffusion coefficients for potassium sorbate and natamycin were in the ranges 1.09 × 10⁻¹¹ to 13.0 × 10⁻¹¹ m²/s and 6.16 × 10⁻¹⁴ to 37.8 × 10⁻¹⁴ m²/s, respectively, and significantly decreased as the WPI-glycerol ratio increased. No significant difference in sorbate diffusion was seen with the addition of BW.     

Technical Approach to Simplify the Purification Method and Characterization of Microbial Transglutaminase Produced from Streptoverticillium ladakanum

In order to fast and economically purify MTGase from Streptoverticillium ladakanum , a stepwise elution method was developed and compared with linear gradient elution method. MTGase was purified to electrophoretical homogeneity by using CM Sepharose CL-6B and Blue Sepharose Fast Flow chromatographies by linear gradient or stepwise methods. The recovery of MTGase by linear gradient and stepwise methods were 68.4% and 81.0%, respectively. The optimal temperature and pH were 40 °C and 5.5, respectively. It was stable at pH 5.0 to 7.0 and had a rate constant (KD) of 6.21 °o 10^-5 min^-1 for thermal inactivation at 45 °C. The purified MTGase was activated by K⁺ Na⁺, Ca²⁺, Mn²⁺, and Mg²⁺, not affected by Fe³⁺, EDTA, but inhibited by Cu²⁺, Zn²⁺, Hg²⁺, Ni²⁺, Co²⁺, Cd²⁺, PCMB, NEM, IAA, and PMSF. A simple stepwise method was developed for the purification of MTGase from S. ladakanum.     

Inactivation of Listeria monocytogenes on Beef Bologna and Cheddar Cheese Using Polyvinyl-idene Chloride Films Containing Sorbic Acid

ABSTRACT: Polyvinylidene chloride (PVDC, Saran^R F-310) films containing sorbic acid (0%, 1.5%, and 3.0% w/v) were prepared with use of a solvent-casting method and were then placed between slices of commercially produced beef bologna that were previously surface-inoculated with L. monocytogenes at 10³ or 10⁵ CFU/g. In addition, cubes of commercial Cheddar cheese were surface-inoculated to contain 10³ or 10⁵ Listeria monocytogenes colony-forming units (CFU) /g and then wrapped with the sorbic acid-containing films. Films containing 1.5% and 3.0% (w/ v) sorbic acid prevented growth of L. monocytogenes on bologna slices with populations as much as 7.1 logs lower after 28 d of storage at 4 °C compared with the sorbic acid-free controls. In contrast, numbers of Listeria remained relatively stable on Cheddar cheese with populations decreasing < 1.3 logs after 35 d of storage. With use of the sorbic acid-containing films, common spoilage organisms were also inhibited on both products. After 28 d of contact with bologna and Cheddar cheese, these films retained 7% and 60% of their original sorbic acid content, respectively, with the control film retaining 85% of its original sorbic acid content. Given these findings, sorbic acid-containing films may be useful in enhancing the safety and shelf-life of ready-to-eat delicatessen products.