Rheological,gelling and emulsifying properties of a glycosylated and cross‐linked caseinate generated by transglutaminase

The impacts of oligochitosan glycosylation and cross‐linking on some properties of a commercial caseinate were investigated in this study. The glycosylated and cross‐linked caseinate with glucosamine content of 4.74 g kg^?1 protein was generated by transglutaminase (TGase) and oligochitosan at pH 7.5 and 37 °C, with fixed substrate molar ratio of 1:3 (acyl donor to glucosamine acceptor), caseinate content of 50 g L^?1, TGase of 10 kU kg^?1 protein and reaction time of 3 h, respectively. In comparison with the caseinate, the glycosylated and cross‐linked caseinate had decreased reactable amino groups (0.58 vs. 0.51 mol kg^?1 protein), higher apparent viscosity, decreased emulsifying activity index (about 14.5%) and statistically unchanged emulsion stability index (92.6 vs. 90.5%). Based on the mechanical spectra of the acid‐induced gels, the glycosylated and cross‐linked caseinate showed shorter gelation time (95 vs. 200 or 220 min) than the caseinate or cross‐linked caseinate. The gels prepared from the glycosylated and cross‐linked caseinate also had enhanced hardness, springiness and cohesiveness. The results indicated that TGase‐mediated oligochitosan glycosylation and cross‐linking has the potential to obtain new protein ingredients.     

The preparation of an oligochitosan‐glycosylated and cross‐linked caseinate obtained by a microbial transglutaminase and its functional properties

A glycosylated and cross‐linked caseinate (GCC) with glucosamine amount of 4.74 g/kg protein was generated from caseinate and oligochitosan by a microbial transglutaminase. The applied temperature, pH and molar ratio of acyl donor/acceptor were 37 °C, 7.5 and 1:3, respectively; while caseinate concentration, transglutaminase addition and reaction time selected from single‐factor trials were 50 g/L, 10 kU/kg protein and 3 h, respectively. Electrophoretic analysis revealed the cross‐linking and glycosylation of caseinate. Compared with caseinate, GCC showed improved solubility in pH 4–11, higher digestibility in vitro and water binding capacity, about 3‐fold, but lower surface hydrophobicity and oil binding capacity (34%).     

Effect of microbial transglutaminase on the functional properties of megrim (Lepidorhombus boscii) skin gelatin

This paper examines the effect of a microbial transglutaminase (TGase) on the gelling and viscoelastic properties of a gelatin from megrim (Lepidorhombus boscii) skins. Although TGase extended the setting time of fish gelatin, it was found that melting temperature, gel strength and viscosity in solution at 60 °C were considerably increased by the covalent cross‐linking action of the enzyme, as observed by SDS‐PAGE and SEM. Increasing concentrations of TGase increased the elasticity and cohesiveness of gelatin gels but reduced gel strength and hardness. Partial inactivation of the enzyme was achieved thermally without negatively affecting the properties of the gelatin; whether or not gelatin is thermoreversible depends essentially on the degree of enzyme inactivation. © 2001 Society of Chemical Industry     

Functional properties of limited hydrolysed cross‐linked casein–gelatin composites

A casein–gelatin composite was prepared by cross‐linking of caseinate and bovine gelatin with a microbial transglutaminase, and the impact of limited proteolysis by trypsin on some functional properties of the composite was investigated in the present work. Two hydrolysed composites were prepared with degree of hydrolysis (DH) of 1 and 2% and analysed by SDS‐PAGE to reflect polypeptide profiles. Some functional properties of the two hydrolysed composites were evaluated, among which solubility, digestibility in vitro, surface hydrophobicity and emulsifying properties showed dependence on the DH. Limited proteolysis of the composite improved its solubility in pH 3–10, especially when the DH was 2%. Compared to the composite, the hydrolysed composites showed an increased emulsifying activity index (about 357–408%), emulsion stability index (about 23–28%) and digestibility in vitro (about 65–80% for pepsin, whereas 2–3% for pepsin–trypsin hydrolysis), together with a decreased surface hydrophobicity (about 55–62%) and oil absorption capacity (about 20–23%). The applied proteolysis also led to the aqueous dispersions of the two hydrolysed composites much lower apparent viscosity, storage and loss modulus.     

Effect of microbial transglutaminase on autolysis and gelation of lizardfish surimi

In the absence of microbial transglutaminase (MTGase), the textural properties of lizardfish surimi (Saurida spp) improved when pre‐incubated at 4 and 25 °C for 24 and 4 h, respectively. MTGase optimally catalyzed incorporation of monodansylcadaverine (MDC) into surimi at 40 °C. Addition of MTGase appeared to reduce autolytic activity at 25 and 40 °C, but had no effect on autolytic activity at 65 °C. Breaking force and deformation of lizardfish surimi significantly improved when 0.1 unit MTGase g^?1 surimi (1.8 g kg^?1) was added and pre‐incubated at either 25 or 40 °C. Textural properties improved concomitant with cross‐linked polymers of myosin heavy chain and tropomyosin, but not actin. Addition of MTGase also improved the storage modulus (G′). The gel network of surimi mixed with MTGase and pre‐incubated at 40 °C readily formed during the pre‐incubation period, while formation of the gel network began at 48.1 °C in the absence of MTGase. Copyright © 2005 Society of Chemical Industry     

Comparative study on protein cross-linking and gel enhancing effect of microbial transglutaminase on surimi from different fish

BACKGROUND: Microbial transglutaminase (MTGase) has been used to increase the gel strength of surimi. Nevertheless, its effectiveness varies with fish species. The aim of this study was to elucidate the effect of MTGase at different levels on protein cross‐linking and gel property of surimi from threadfin bream, Indian mackerel and sardine in the presence and absence of endogenous transglutaminase. RESULT: Breaking force of all surimi gels increased as MTGase levels (0–0.6 U g^?1) increased except for threadfin bream surimi gel, where the breaking force decreased at 0.6 U g^?1 (P < 0.05). In the presence of EDTA, the gel strengthening effect was lower, suggesting the combined effect of endogenous transglutaminase with MTGase. With the addition of MTGase, the gel with the highest increase in breaking force showed highest decrease in myosin heavy chain. When cross‐linking activity of MTGase on natural actomyosin (NAM) was determined, the highest decreasing rate in ε‐amino group content with the concomitant increased formation of cross‐linked proteins was found in NAM from threadfin bream. The reactivity of muscle proteins toward MTGase‐induced cross‐linking was in agreement with surimi gel strengthening. CONCLUSION: The composition and properties of muscle proteins of varying fish species more likely determined protein cross‐linking induced by MTGase, thereby affecting their gel properties. Copyright © 2011 Society of Chemical Industry     

Zinc‐loaded whey protein nanoparticles prepared by enzymatic cross‐linking and desolvation

Zinc‐loaded whey protein nanoparticles were prepared by enzymatic cross‐linking whey protein followed by ethanol desolvation. Whey protein isolate (WPI) was denatured by heating (80 °C for 15 min) at pH 7.0 and then cross‐linked by transglutaminase at 50 °C for 4 h while stirring. Transglutaminase was inactivated by heating at 90 °C for 10 min, and then, ZnSO₄·7H₂O (1–10 mm ) was added. Zinc‐loaded whey protein nanoparticles were formed by adding ethanol at one to five times the volume of the protein solution at pH 9.0. The desolvated solutions were diluted by adding distilled water at ratio of 1:100 (w/v) immediately after desolvation. Dynamic light scattering (DLS) data showed that the particle size of zinc‐loaded whey protein nanoparticles increased with the amount of zinc and the volume of ethanol. Scanning electron microscopy micrographs revealed an almost spherical morphology for zinc‐loaded whey protein nanoparticles. The zinc loading efficiency was determined ranging from 76.7% to 99.2%. In vitro test data showed that the zinc release rate was low in simulated gastric fluid but high in simulated intestinal fluid. The results indicated that enzymatic cross‐linked whey protein nanoparticles may be used as a good vehicle to deliver zinc as a supplement.     

Effect of microbial transglutaminase on gel properties and film characteristics of gelatin from lizardfish (Saurida spp.) scales

The addition of microbial transglutaminase (MTGase) generally increased the gel strength of lizardfish (Saurida spp.) scale gelatin gels (P≤0.05) with an increase in gel strength with the addition of MTGase up to 0.5% (w/v). The texture profile analysis compression tests of lizardfish scale gelatin gel with and without MTGase were studied to determine their effects on gel characteristics. MTGase added to the gels decreased the band intensity of the β- and α-components with increasing concentrations of enzyme. Gel microstructures with various concentration of MTGase showed denser strands in the gels with enzyme compared with the looser stands in non-enzyme-treated gel samples. Films cast from lizardfish scale gelatin with and without 0.5% MTGase and bovine gelatin films were transparent and flexible. The lizardfish gelatin films were all slightly yellowish while the bovine gelatin films were clearer. The L value of bovine gelatin films had the highest value (P≤0.05) whereas lizardfish scale gelatin films with and without enzyme were not significantly different (P>0.05) for L, a, and b values and ΔE. The film's mechanical properties included tensile strength (TS) and elongation at break (E) were not significantly different (P > 0.05) for E and the films of lizardfish scale gelatin showed higher TS than the films without enzyme added (P ≤ 0.05). The water vapor permeability of films from lizardfish scale gelatin with and without 0.5% MTGase and bovine gelatin films were 21.0 ± 0.17, 26.3 ± 0.79, and 25.8 ± 0.09 g·mm/m(2)·d·kPa, respectively, while the oxygen transmission rate of all 3 types of films were less than 50 cc O(2)/m(2)·d.     

酪蛋白的转谷氨酰胺酶酶促糖基化交联条件及产物性质

在氨基葡萄糖存在的条件下,利用转谷氨酰胺酶(EC2.3.2.13)对酪蛋白进行糖基化交联修饰。以修饰酪蛋白产物中氨基葡萄糖的导入量为指标,采用单因素试验分别考察反应体系pH值、酶添加量、反应温度和时间对修饰反应的影响。优化后的适宜修饰条件为:酪蛋白底物质量浓度为30g/L,氨基葡萄糖添加量为3mol(每kg酪蛋白中)、pH值为7.5、酶添加量为10kU(每kg酪蛋白中)、反应温度为37℃、时间4h。与酪蛋白和转谷氨酰胺酶促交联的酪蛋白相比,修饰酪蛋白产物的乳化性质和胶凝性质得到显著改善,并且体外消化性能未受到影响,表明转谷氨酰胺酶催化的糖基化交联修饰可以用于改善酪蛋白的这些功能性质。     

Effect of a microbial calcium‐independent transglutaminase on functional properties of a partially purified cowpea (vigna unguiculata) globulin

A partially purified globulin from cowpea seeds was polymerised by calcium‐independent microbial transglutaminase. The level of free amino groups in the globulin decreased with increase in enzyme concentration, suggesting that the cross‐linking reaction involved participation of the amino residues. At transglutaminase concentrations of 6–18 μg ml⁻¹ emulsifying activity decreased with increasing enzyme concentration and degree of cross‐linking, while a similar trend was observed for the foaming property at 6–12 μg ml⁻¹ transglutaminase concentrations. The resultant emulsions and foams formed by the cross‐linked proteins were more stable than those formed by the untreated protein. Sodium dodecyl sulphate polyacrylamide gel electrophoresis showed that of the two major polypeptides, the 55 kDa protein was more susceptible to transglutaminase action than the 50 kDa protein. Scanning electron microscopy revealed an amorphous structure for the control protein gel while a more defined and cross‐linked gel structure was observed for the protein treated with transglutaminase. The results suggest that protein products differing in functionalities can be obtained by controlling the degree of enzymatic protein cross‐linking. © 1999 Society of Chemical Industry     

Gel‐enhancing effect and protein cross‐ linking ability of tilapia sarcoplasmic proteins

BACKGROUND: Tilapia (Oreochromis niloticus) sarcoplasmic proteins contain substantial transglutaminase (TGase) activity. The enzyme catalyzes the protein cross‐linking reaction, resulting in a more elastic gel. The objective was to investigate the gel‐enhancing effect of sarcoplasmic proteins from tilapia as related to TGase activity. RESULTS: Total TGase activity of sarcoplasmic proteins concentrate (SpC) increased about 3.6‐fold after ultrafiltration using 30 kDa membrane, but specific activity remained unchanged, indicating minimal TGase purification by ultrafiltration. Addition of 1 mg mL^?1 SpC containing 40 units TGase activity induced cross‐linking of tilapia actomyosin, and the extent of cross‐linking increased with added level of SpC. Myosin heavy chain (MHC) and troponin were preferably cross‐linked by tilapia SpC, while actin and tropomyosin were not affected. Higher retention of MHC was observed concomitantly with greater content of cross‐linked protein when SpC was added to lizardfish surimi. Lizardfish surimi with 10 g kg^?1 SpC added and pre‐incubated at 37 °C for 1 h exhibited 91.6% and 26.7% increase in breaking force and deformation, respectively, when compared to the control. CONCLUSIONS: Residual TGase activity in SpC played an important role in catalyzing the protein cross‐linking and enhancing actomyosin gelation. SpC could be a potential ingredient for improving textural properties of fish protein gel. Copyright © 2007 Society of Chemical Industry     

Tuning the Functional Properties of Polysaccharide–Protein Bio‐Based Edible Films by Chemical,Enzymatic, and Physical Cross‐Linking

Among natural biopolymers, polysaccharides and proteins are very promising for biodegradable and edible wraps with different characteristics, so that their formulations can be tailor‐made to suit the needs of a specific commodity. Films prepared from polysaccharides have good gas barrier properties but exhibit lower resistance to moisture compared to protein films (edible) or polylactide films (biodegradable). Protein‐based films show better mechanical and oxygen barrier properties compared to polysaccharide films. For that reason, film performances may be enhanced by producing blend systems, where hydrocolloids (mixtures of proteins and/or polysaccharides) form a continuous and more cohesive network. However, the lower water barrier properties of hydrocolloid films and their lower mechanical strength in comparison with synthetic polymers limit their applications in food packaging. Therefore, the enhancement of biopolymer film properties has been studied to attain appropriate applications. This review provides an extensive synthesis of the improvement of the properties of edible polysaccharide–protein films by way of various chemical, enzymatic, and physical methods. These methods primarily aim at improving the mechanical resistance. They also permit to ameliorate the water and gas barrier properties and related functional properties.     

Production of a transglutaminase from Zea mays in Escherichia coli and its impact on yoghurt properties

A gene from Zea mays coding a TGase was expressed in E. coli and identified by Western blotting. Under optimal expression conditions, the production and specific activity of refolded TGZ were 1.41 mg/L and 0.34 U/mg. The activated TGZ was employed to cross‐link milk proteins. The yoghurt treated with TGZ showed a lower syneresis, higher apparent viscosity and texture than untreated yoghurt. The properties of TGZ‐treated sample were better than those of MTG‐treated samples, so TGZ may have potential as an additive in yoghurt manufacture.     

Comparison of gelling properties and flow behaviors of microbial transglutaminase (MTGase) and pectin modified fish gelatin

The gelling and structural properties of microbial transglutaminase (MTGase) and pectin modified fish gelatin were compared to investigate their performances on altering fish gelatin properties. Our results showed that within a certain concentration, both MTGase and pectin had positive effects on the gelation point, melting point, gel strength, textural, and swelling properties of fish gelatin. Particularly, low pectin content (0.5%, w/v) could give fish gelatin gels the highest values of gel strength, melting temperature, and hardness. Meantime, flow behavior results showed that both MTGase and pectin could increase fish gelatin viscosity without changing its fluid characteristic, but the latter gave fish gelatin higher viscosity. Both MTGase and pectin could increase the lightness of fish gelatin gels but decreases its transparency. More importantly, fluorescence and UV absorbance spectra, particle size distribution, and confocal microscopy results indicated that MTGase and pectin could change the structure of fish gelatin with the formation of large aggregates. Compared with MTGae modified fish gelatin, pectin could endow fish gelatin had similar gel strength, thermal and textural properties to pig skin gelatin.