Modulation of mechanical and surface hydrophobic properties of food protein films by transglutaminase treatment

The effects of microbial transglutaminase (MTGase) treatment on the mechanical and hydrophobic properties of cast films from various food proteins were investigated. SDS–PAGE analyses confirmed that the MTGase treatment led to the formation of insoluble film network for most of food proteins. This enzymatic treatment significantly (P < 0.05) increased tensile strength (TS) values of cast films of all food proteins by 13–33% (except whey proteins). Meanwhile, the influence on the elongation at break (EB) values was mainly dependent upon the molecular nature of proteins. For most of food proteins, both increases in TS and EB values of films were obtained after the MTGase treatment. Furthermore, the surface hydrophobicity of most of protein films was also significantly improved. These results suggest that the enzymatic treatment by MTGase could be used as an effective technique to improve mechanical and surface hydrophobic properties of protein films.     

Effects of transglutaminase treatment on the thermal properties of soy protein isolates

The effects of covalent cross-linking of microbial transglutaminase (MTGase) on the thermal properties of soy protein isolates (SPI), including the thermal denaturation and glass transition were investigated by conventional and modulated differential scanning calorimetry (DSC). The MTGase treatment significantly increased the thermal denaturation temperatures (including the on-set temperature of denaturation, T_m and the peak temperature of denaturation, T_d) of glycinin and β-conglycinin components of SPI (P ⩽ 0.05), and the thermal pretreatment of SPI further increased the extent of this improvement. The MTGase treatment also improved the ability of SPI to resist the urea-induced denaturation. Modulated DSC analysis showed that there were two glass transition temperatures (T_g) in the reversible heat flow signals of native SPI (about 5% moisture content), approximately corresponding to 45 and 180 °C, respectively. These T_g values of SPI were significantly decreased by the MTGase treatment (at 37 °C for more than 2 h) (P ⩽ 0.05). The improvement in the hydration ability of protein and the formation of high molecular biopolymers may account for the changes of thermal properties of soy proteins caused by the MTGase cross-linking.     

Gelling of microbial transglutaminase cross-linked soy protein in the presence of ribose and sucrose

Soy protein isolate (SPI) was incubated with microbial transglutaminase (MTGase) enzyme for 5 (SPI/MTG(5)) or 24 (SPI/MTG(24)) h at 40 °C and the cross-linked SPI obtained was freeze-dried, and heated with 2% (w/v) ribose (R) for 2 h at 95 °C to produce combined-treated gels. Longer incubation period resulted in more compact and less swollen SPI particle shape when reconstituted with sugar solution. Thus, this MTGase treatment affected samples in terms of flow behaviour and gelling capacity. Rheological study showed different gelling profiles with the cross-linking treatments and combined cross-linked SPI gave a higher G′ value compared to single treated samples. These are due to the formation of additional ε-(γ-glutamyl)lysine bonds and “Maillard cross-links” within the SPI protein network during the MTGase incubation and heating in the presence of ribose (i.e. reducing sugar). Network/non-network protein analysis found that network protein increased with cross-linking treatment, which also resulted in different SDS–PAGE profiles. As in non-network protein fraction, A₄ subunit was suggested to become part of the network protein as a result of combined cross-linking.     

Effect of soy protein substitution for sodium caseinate on the transglutaminate-induced cold and thermal gelation of myofibrillar protein

The influence of soy protein isolate (SPI) substitution for sodium caseinate (SC) on the properties of cold-set (4 °C) and heat-induced gels of pork myofibrillar protein (MP) incubated with microbial transglutaminase (TG) was investigated. The strength of cold-set MP–SC gels (formed in 0.45 M, NaCl, 50 mM phosphate buffer, pH 6.25) increased with time of TG incubation, but those gels with more than 66% SPI substituted for SC had a >26% reduced strength (P < 0.05). Upon cooking, both incubated and non-incubated protein sols were quickly transformed into highly elastic gels, showing up to 6000 Pa in storage modulus (G′) at the final temperature (72 °C). However, no differences (P < 0.05) in G′ were observed between heated samples with SPI and SC. Myosin heavy chain, casein and soy proteins gradually disappeared with TG incubation, contributing to MP gel network formation. Both cold-set and heat-induced gels had a compact protein matrix, attributable to protein cross-linking by TG.     

Incorporation of microbial transglutaminase into non-fat yogurt production

This study investigated the physical, chemical and sensory characteristics of non-fat yogurts treated with microbial transglutaminase (MTGase) at varying concentrations from 0 to 0.5 g L⁻¹. Also, the effect of enzyme inactivation prior to fermentation on the selected properties of the yogurts was studied. Acid development rate was reduced with increasing MTGase doses. Cross-linking of milk proteins by MTGase had a growth-slowing effect on yogurt starter bacteria, which was more pronounced at higher concentrations. Physical properties of the yogurts were improved by MTGase throughout 21-day storage; on the contrary, the production of acetaldehyde was slowed down by increasing MTGase concentrations during the same period. Principal component analysis (PCA) and hierarchial cluster analysis (HCA) clearly differentiated the samples with added MTGase at lower (⩽0.3 g L⁻¹) and higher (0.4–0.5 g L⁻¹) concentrations regarding the physical and sensory properties. The physical and sensory properties of non-fat set yogurt could be improved by incorporating MTGase up to a level of 0.3 g L⁻¹.     

Formation and properties of glycinin-rich and β-conglycinin-rich soy protein isolate gels induced by microbial transglutaminase

The gelation and gel properties of glycinin-rich and β-conglycinin-rich soy protein isolates (SPIs) induced by microbial transglutaminase (MTGase) were investigated. At the same enzyme and protein substrate concentrations, the on-set of gelation of native SPI and the viscoelasticity development of correspondingly formed gels depended upon the relative ratio of glycinin to β-conglycinin. The turbidity analysis showed that the glycinin components also contributed to the increase in the turbidity of SPI solutions incubated with MTGase (at 37 °C). Textural profile analysis indicated that the glycinin components of SPIs principally contributed to the hardness, fracturability, gumminess and chewiness values of corresponding gels, while the cohesiveness and springness were mainly associated with the β-conglycinin components. The strength of MTGase-induced gels of various kinds of SPIs could be significantly improved by the thermal treatment. The protein solubility analyses of MTGase-induced gels, indicated that besides the covalent cross-links, hydrophobic and H-bondings and disulfide bonds were involved in the formation and maintenance of the glycinin-rich SPI gels, while in β-conglycinin-rich SPI case, the hydrophobic and H-bondings were the principal forces responsible for the maintenance of the gel structure. The results suggested that various kinds of SPI gels with different properties could be induced by MTGase, through controlling the glycinin to β-conglycinin ratio.     

The effect of pH on the gelling behaviour of canola and soy protein isolates

The present study investigates the gelation mechanisms of a canola protein isolate (CPI) as a function of a pH (3.0–9.0), and compares it to that of a commercial soy protein isolate (SPI). A rheological investigation found that CPI was non-gelling at pH 3.0, and then formed a gel with increasing strength as pH was raised from pH 5.0 to 9.0. In contrast, the commercial SPI ingredient was found to be non-gelling at pH 9.0, but formed the strongest networks at pH 5.0 near its isoelectric point (pI = 4.6). Denaturation temperature as determined by differential scanning calorimetry were found to occur at ~ 78 °C for CPI at pH 5.0, then shifted to higher temperatures (~ 87 °C) at pH 7.0/9.0, whereas detection of SPI denaturation could not be obtained due to instrument sensitivity. Gelling temperatures were similar for both CPI and SPI (~ 82–86 °C) at all pHs, with the exception of SPI at pH 5.0 (~ 46 °C). Overall CPI networks were stronger than SPI, since the latter had weaker inter- and intramolecular junction zones. Confocal laser scanning microscopy images indicated that CPI gels became denser with lower lacunarity values as pH increased from 3.0 to 9.0. Moreover, the fractal dimension of CPI gels was found to increase from ~ 1.5-1.6 to ~ 1.8 as pH increased from 5.0/7.0 to 9.0, respectively suggesting diffusion-limited cluster-cluster aggregation. Images of SPI networks were not concurrent with fractal analysis under the conditions examined. Despite CPI having excellent gelling properties that are comparable to SPI, its need for alkaline pH conditions will limit its applicability in foods.     

Edible films produced with gelatin and casein cross-linked with transglutaminase

In this study, transglutaminase was used to produce cross-linked casein, gelatin and casein–gelatin blend (100:0, 75:25, 50:50, 25:75 and 0:100) edible films. Cross-linking was investigated by SDS–PAGE. Mechanical and water vapor barrier properties of the films were characterized using ASTM procedures, and the film morphology was evaluated using scanning electron microscopy. The casein–gelatin film showed significant greater elongation values (P < 0.05) with or without transglutaminase treatment, as compared to films made from gelatin or casein alone. Mixtures of casein and gelatin produced a synergistic effect only observed in the film elongation, while no improvement was detected for tensile strength and water vapor barrier properties, except for the casein:gelatin (75:25) formulation with transglutaminase, which showed the lowest water vapor permeability value (5.06 ± 0.31 g mm/m² d kPa). Enzymatic cross-linking also induced a substantial increase in the high molecular weight protein components in the film forming solutions.     

Features and performance of edible films,obtained from whey protein isolate formulated with antimicrobial compounds

The goal of this research effort was to assess the efficacy of edible films produced from whey protein isolate (WPI) and glycerol, including incorporation of lactic acid (LA) and propionic acid (PRO), chitooligosaccharides with nominal MW of 3 kDa (COS) and natamycin (NA) as antimicrobial agents. Their features were evaluated in vitro via agar diffusion and viable cell counting, against spoilage microflora often found contaminating cheese surfaces. The effect of incorporating the aforementioned compounds upon thickness, moisture content (MC), solubility (S), density (ρ^s), water activity (a_w) and water vapor permeability (WVP), as well as upon tensile and optical properties of those films were also evaluated. Films formulated with LA, PRO or COS exhibited antimicrobial activity against all microorganisms tested, yet the viable cell count assay was more sensitive and reproducible. COS was the most active against Gram-negative bacteria, whereas LA was the most active against Gram-positive ones. NA was not active against bacteria, but displayed the strongest effect against yeasts. Incorporation of said antimicrobial compounds did not significantly (p > 0.05) affect film thickness, yet it significantly (p < 0.05) reduced tensile strength (TS). Incorporation of LA and NA in particular did not significantly (p < 0.05) affect MC, S, ρ^s, WVP, elongation at break (EB) and Young's modulus (YM) values; however, a statistically significant increase (p < 0.05) of MC, S and WVP, together with a statistically significant decrease (p < 0.05) of ρ^s were attained upon incorporation of PRO or COS. Moreover, PRO produced the highest variation (p < 0.05) in EB, TS and YM, whereas COS produced the highest change (p < 0.05) in optical properties.     

Preparation and functional properties of fish gelatin–chitosan blend edible films

With the goal of improving the physico-chemical performance of fish gelatin-based films, composite films were prepared with increasing concentrations of chitosan (Ch) (100G:0Ch, 80G:20Ch, 70G:30Ch, 60G:40Ch and 0G:100Ch, gelatin:Ch), and some of their main physical and functional properties were characterised. The results indicated that the addition of Ch caused significant increase (p < 0.05) in the tensile strength (TS) and elastic modulus, leading to stronger films as compared with gelatin film, but significantly (p < 0.05) decreased the elongation at break. Ch drastically reduced the water vapour permeability (WVP) and solubility of gelatin films, as this decline for the blend film with a 60:40 ratio has been of about 50% (p < 0.05). The light barrier measurements present low values of transparency at 600 nm of the gelatin–chitosan films, indicating that films are very transparent while they have excellent barrier properties against UV light. The structural properties investigated by FTIR and DSC showed a clear interaction between fish gelatin and Ch, forming a new material with enhanced mechanical properties.     

Quercetagetin loaded in soy protein isolate–κ-carrageenan complex: Fabrication mechanism and protective effect

In the present study, the potential of soy protein isolate (SPI)–κ-carrageenan (κ-CG) complex as a protective carrier for quercetagetin was investigated at different pH values (pH 2.3 and 6.5). The particle size of the ternary aggregates was slightly increased at pH 2.3, yet dramatically decreased at pH 6.5 with increasing quercetagetin concentration. Moreover, the negative ζ-potential of the ternary aggregates was increased significantly (p < 0.05) at pH 6.5. The addition of quercetagetin to the SPI–κ-CG complex could highly quench the intrinsic fluorescence of SPI. Circular dichroism spectra further suggested that the bound quercetagetin could induce the rise of β-sheet and β-turn contents at the cost of α-helix and unordered coil fractions of SPI. In addition, quercetagetin could increase the viscoelasticity of the ternary aggregates at both pH. Furthermore, the SPI–κ-CG complex was found to be superior to single SPI or κ-CG in terms of improving light stability and radical scavenging ability of quercetagetin.     

Ultra-High Pressure Homogenization enhances physicochemical properties of soy protein isolate-stabilized emulsions

The effect of Ultra-High Pressure Homogenization (UHPH, 100–300 MPa) on the physicochemical properties of oil-in-water emulsions prepared with 4.0% (w/v) of soy protein isolate (SPI) and soybean oil (10 and 20%, v/v) was studied and compared to emulsions treated by conventional homogenization (CH, 15 MPa). CH emulsions were prepared with non-heated and heated (95 °C for 15 min) SPI dispersions. Emulsions were characterized by particle size determination with laser diffraction, rheological properties using a rotational rheometer by applying measurements of flow curve and by transmission electron microscopy. The variation on particle size and creaming was assessed by Turbiscan® analysis, and visual observation of the emulsions was also carried out. UHPH emulsions showed much smaller d_3.2 values and greater physical stability than CH emulsions. The thermal treatment of SPI prior CH process did not improve physical stability properties. In addition, emulsions containing 20% of oil exhibited greater physical stability compared to emulsions containing 10% of oil. Particularly, UHPH emulsions treated at 100 and 200 MPa with 20% of oil were the most stable due to low particle size values (d_3.2 and Span), greater viscosity and partial protein denaturation. These results address the physical stability improvement of protein isolate-stabilized emulsions by using the emerging UHPH technology.     

Effect of microbial transglutaminase treatment on thermal stability and pH-solubility of heat-shocked whey protein isolate

Whey protein isolate (WPI) dispersions (5% protein, pH 7.0) were subjected to heat-shock at 70 °C for 1, 5 and 10 min. The heat-shocked WPI dispersions were treated with microbial transglutaminase (MTGase) enzyme, and thermal properties and pH-solubility of the treated proteins were investigated. Heat-shocking of WPI for 10 min at 70 °C increased the thermal denaturation temperature (T_d) of β-lactoglobulin in WPI by about 1.5 °C. MTGase treatment (30 h, 37 °C) of the heat-shocked WPI significantly increased the T_d of β-lactoglobulin by about 6.3–7.3 °C when compared with heat-shocked only WPI at pH 7.0. The T_d increased by about 13–15 °C following pH adjustment to 2.5; however, the T_d of heat-shocked WPI was not substantially different from heat-shocked and MTGase-treated WPI at pH 2.5. Both the heat-shocked and the heat-shocked-MTGase-treated WPI exhibited U-shaped pH-solubility profiles with minimum solubility at pH 4.0–5.0. However, the extent of precipitation of MTGase-treated WPI samples at pH 4.0–5.0 was much greater than all heat-shocked and native WPI samples. The study revealed that while MTGase cross-linking significantly enhanced the thermal stability of β-lactoglobulin in heat-shocked WPI, it caused pronounced precipitation at pH 4.0–5.0 via decreasing the hydrophilic/hydrophobic ratio of the water-accessible protein surface.     

Effects of ribose, microbial transglutaminase and soy protein isolate on physical properties and in-vitro starch digestibility of yellow noodles

Soy protein isolate (SPI), microbial transglutaminase (MTGase) and ribose (R) were used to modify physical properties and in-vitro starch hydrolysis of yellow noodle. Four types of noodles were produced; noodles with SPI (SPI/C noodles), noodles with SPI and ribose (SPI/R noodles), noodles with SPI and microbial transglutaminase (SPI/MTGase noodles) and noodles with SPI, ribose and MTGase (SPI/R/MTGase noodles). γ-glutamyl-lysine bonds by MTGase and ribose-induced Maillard reaction within SPI were induced by incubating the noodles for 5 h at 40 °C followed by steaming for 30 min. Cooked noodles were assessed for physical properties such as pH, color, tensile strength and elasticity, and in-vitro hydrolysis index (HI) and estimated glycemic index (GI). SPI/R/MTGase and SPI/MTGase noodles exhibited significantly (P < 0.05) higher tensile strength and elasticity than SPI/R and SPI/C noodles. HI and GI were in the order; SPI/R/MTGase < SPI/MTGase < SPI/R < SPI/C noodles. Incorporation of SPI that was treated with MTGase and ribose may be useful for controlling the texture and starch hydrolysis of yellow noodles. These attributes may be due to the formation of γ-glutamyl-lysine bonds during incubation of SPI, and ribose-induced Maillard reaction during steaming of the noodles.     

Oxygen permeability and mechanical properties of banana films

Banana flour, obtained from banana cv. Kluai Namwa, was used to form banana films. The effect of banana flour, glycerol (Gly) and pectin content on film oxygen permeability (OP) and mechanical properties of banana films were studied. Banana flour content significantly affected film OP; whereas, Gly and pectin contents did not significantly affect film OP. Increasing banana flour and pectin contents enhanced film strength; thus, it showed higher film elastic modulus (EM) and tensile strength (TS) but less film % elongation (% E) (p ⩽ 0.05). In contrast, increasing Gly content reduced the film strength and improved film flexibility; therefore, it decreased film EM and TS but increased film % E (p ⩽ 0.05). Banana films showed good sealability, which can make these films suitable as sachets or pouches for dry foods, thus reducing the need for plastic materials.     

Enzymatic Production of Ethanol from Cassava Starch Using Two Strains of Saccharomyces cerevisiae

Cassava starch from TMS 30572 and Idileru were hydrolyzed with α-amylase and amylo-glucosidase before fermentation using two strains of Saccharomyces cerevisiae from palm wine and bakers’ yeast. The per cent yield of sugars and total dissolved solids were 66 % and 26% respectively while pH was 7. Spectrophotometric measurement of the cell growth revealed steady but insignificant (p ≤ 0.05) increase in cell concentrations up to 48 h fermentation time with a gradual decline by 72 h. Saccharomyces cerevisiae strain from palm wine grew best on TMS 30572 hydrolysate at 20% sugar concentration (optical density 0.663; fermentation time 48 h) while on Idileru hydrolysate it grew best at 25% (optical density 0.698; fermentation time 60 h). The pH values obtained from the fermenting hydrolysates for both yeast strains declined gradually as the fermentation progressed with the lowest pH values (3.01 for S. cerevisiae from palm wine; 3.06 for S. cerevisiae from bakers’ yeast) obtained for TMS 30572 cassava variety at 25% sugar concentration. Changes in pH were significant (p ≤ 0.05). The Saccharomyces cerevisiae strain from palm-wine had a higher conversion of available sugar into ethanol. The yield of ethanol was found to vary but the highest ethanol concentration obtained was 5.3% at 10% initial sugar concentration, which gave a sugar conversion efficiency of 37.3%. The results obtained suggest that Saccharomyces cerevisiae strains from sources other than those used conventionally can serve as good substitutes for bio-conversion processes in the industrial production of ethanol.     

An exploratory study on the influence of orange juice on gut microbiota using a dynamic colonic model

The aim of this research was to evaluate the influence of fresh orange juice (FOJ) and pasteurized orange juice (POJ) on gut microbiota using the Simulator of the Human Intestinal Microbial Ecosystem (SHIME®) in a long-term experiment. SHIME® vessels were used to investigate orange juice fermentation throughout the colon and to assess changes in microbial composition and fermentation metabolites (short-chain fatty acids, or — SCFA, and ammonium). Antioxidant activity of the SHIME® vessels and juice was also evaluated. The FOJ increased (p ≤ 0.05) Lactobacillus spp., Enterococcus spp., Bifidobacterium spp., and Clostridium spp. and reduced (p ≤ 0.05) enterobacteria. The POJ increased (p ≤ 0.05) Lactobacillus spp. and reduced (p ≤ 0.05) enterobacteria. The PCR-DGGE analysis showed a reduction in total bacteria population richness values. The FOJ and POJ increased (p ≤ 0.05) butyric, acetic, and propionic acid concentrations, whereas ammonium production was reduced. High values of antioxidant activity were observed as a result of the FOJ and POJ treatments. Principal component analysis indicated that both POJ and FOJ juices had a positive influence on gut microbiota. The FOJ and POJ were found to exhibit selective prebiotic activity, particularly in terms of gut microbiota. This finding is in agreement with increases in both SCFAs and commensal bacteria, as well as with decreases in ammonium levels, though total bacteria richness values were reduced.     

Effect of hydrophobic plasticizers on functional properties of gelatin-based films

The aim of this study was the production and characterization of gelatin-based films using hydrophobic plasticizers derived from citric acid and soy lecithin as emulsifier. The films were characterized as to their mechanic properties, permeability to water vapor, opacity, morphology and possible interactions using Fourier transform infrared spectroscopy. Tensile strength values (TS) varied from 36 to 103 MPa, however, the increase in the concentration of plasticizers (acetyltributyl citrate and tributyl citrate) reduced TS by 57% and no relation was observed between plasticizer quantities and the elongation in the quantities tested. Permeability to water vapor varied between 0.17 and 0.34 (g mm/m² h kPa), slightly increasing with the increase in concentration of plasticizers. The effectiveness in the use of soy lecithin emulsifier in the homogenization between the compounds could be proven by microscopic observation using confocal laser microscopy.     

Biochemical changes throughout the ripening of a traditional Spanish goat cheese variety (Babia-Laciana)

The physico-chemical characteristics, proteolysis (classical nitrogen fractions, caseins and their degradation products and free amino acids), and lipolysis (fat acidity and free fatty acids) were studied throughout the ripening of three batches of Babia-Laciana cheese, a Spanish traditional variety made from raw goats’ milk. The main compositional characteristics of this cheese at the end of the ripening are its high content of total solids (TS) (78.0±2.4 g 100 g⁻¹ of cheese) and fat (61.1±1.2 g 100 g⁻¹ of TS), the presence of residual lactose (1.6±0.8 g 100 g⁻¹ of TS) and its low content of sodium chloride (1.1±0.7 g 100 g⁻¹ of TS) and ash (2.8±0.5 g 100 g⁻¹ of TS). Its pH values (4.44±0.72) are extraordinarily low. The evolution and final values of the different nitrogen fractions show that this cheese undergoes a very slight proteolysis, a fact which was corroborated when the caseins and their degradation products were quantified: β-casein did not undergo any modification throughout ripening, while only 21% of the α_s-caseins were degraded. Free amino acids content increased by a factor of about 7 throughout ripening, resulting in a high content of γ-amino butyric acid and a low content of glutamic acid at the end of the process. Fat acidity increased very slightly, approximately 4.5 times, during ripening, reaching final values of 3.5±2.2 mg KOH g⁻¹ of fat. The total free fatty acids content showed a similar evolution to fat acidity. At the end of the ripening process, the main free fatty acid was C_18:1, followed by C₁₆ and C₁₀.     

Characterization of ‘wet’ alginate and composite films containing gelatin,whey or soy protein

The following study explored how the addition of various proteins (gelatin, soy protein isolate (SPI) and heated/unheated whey protein isolate (WPI)), at two different concentration levels (1% and 2%), affected the mechanical, microstructural and optical properties of calcium cross-linked ‘wet’ alginate films. Additionally, the water holding capacity and textural profile analysis (TPA) properties were determined for the alginate–protein gels. Adding all types of protein significantly (P < 0.05) decreased the force to puncture the ‘wet’ alginate–protein composite films compared to the control alginate film. The tensile test showed significant differences in tensile strength between the various films but interestingly there was no significant difference in the percent elongation at breaks between any of the films. Micrograph images showed that the SPI and heated WPI formed relatively larger protein clumps/regions in the alginate films whereas the gelatin and unheated WPI appeared to be more integrated into the alginate film. The heated WPI films were the least transparent of all the films, followed by the SPI films. Few TPA differences existed between the alginate–protein gels. However, the alginate–gelatin gels did have significantly less water loss than the other alginate–protein gels suggesting that alginate and gelatin may be the most compatible of all the alginate–protein combinations tested.