Application of Transglutaminase in Developing Cassava-based Wet Noodle for Quality and Shelf Life Improvement: A Review.

Characteristic of cassava flour is relatively similar to wheat flour. Cassava flour has the potential to substitute 70-80% of wheat flour as the main ingredient for wet noodle production. Unfortunately, cassava flour has no gluten and lower protein content than wheat flour, which is important for the characteristic of a wet noodle. Therefore, transglutaminase (MTGase) is often applied in non-gluten products to improve its texture. This enzyme catalyzes the reaction between lysine and glutamine to form isopeptide cross-links. Moreover, the addition of MTGase to cassava-based wet noodle improves its texture and color. In addition, this effect gives better palatability for wet noodle. This enzyme can increase the shelf life of wet noodles and safe for our health. The present study demonstrates with patent and literature data the potential of MTGase in noodles based on cassava flour.

Efeitos da transglutaminase na textura de fiambres elaborados com carne suína PSE e teor reduzido de sal / Effects of transglutaminase addition on the texture of ‘fiambres’ elaborated with pork PSE and salt reduction

Objetivou-se avaliar o perfil de textura instrumental e sensorial de fiambres elaborados com transglutaminase (TG), carne suína PSE e redução de sal. Seis formulações com carnes normais (RFN) e PSE foram formuladas contendo 2% de sal, sem sal adicionado e sem sal + TG. A remoção do sal da formulação resultou (P<0,05) em menores valores de dureza e mastigabilidade, independentemente da carne utilizada. A TG foi capaz de evitar a redução nos valores de dureza e mastigabilidade quando os produtos foram elaborados com carne RFN, mas não quando elaborado com carne PSE. A aceitação sensorial da textura foi reduzida com a redução de sal, mas este foi minimizado pela adição da TG. Concluiu-se que apesar da TG minimizar os problemas de textura oriundos da redução de sal nos fiambres, a percepção sensorial não foi favorecida.

Microbial transglutaminase in noodle and pasta processing.

Nowadays, there is an aggressive rate in consumption of noodles and pasta products throughout the world. Consumer acceptability and preference of these functional products can be promoted by the discovery of novel knowledge to improve their formulation and quality. The development of fortified-formulations for noodles and pasta products based on microbial transglutaminase (MTGase) can guarantee the shelf life extension with minimum quality losses. The current review focuses on recent trends and future prospects of MTGase utilization in the structural matrix of noodles and pasta products and represents the quality changes of cooking loss, texture, microstructure, color and sensory attributes of the MTGase-incorporated products. Digestibility, nutritional and health aspects of the MTGase-enriched formulations are also reviewed with a vision toward physical functions and safety outcomes of MTGases isolated from new microbial sources. The high potential of MTGase in developing commercial noodles and pasta products is successfully demonstrated. MTGase by modifying the crystallinity or molecular structure via covalent crosslinks between protein molecules strengthens the doughs stability and the textural characteristics of final products with the low- or high-protein flour. Compared with the control samples, the MTGase-supplemented products indicate slower digestion rates and better sensory and cooking properties without any remarkable color instability.

Improvement of viability of probiotic bacteria, organoleptic qualities and physical characteristics in kefir using transglutaminase and xanthan.

BACKGROUND: Using kefir as a probiotic food carrier has many benefits. At the same time, it is considered an appropriate product for the dairy industry. The aim of this study was to evaluate the effect of xanthan gum and transglutaminase enzyme on the viability of probiotics and the organoleptic qualities and physicochemical characteristics of kefir. METHODS: Three levels of transglutaminase enzyme (50, 100 and 150 ppm), and xanthan gum (0.05%, 0.1% and 0.2%) were used. Sensory and physicochemical properties and viability of probiotic bac- teria were measured over 2 weeks of storage at 4°C. RESULTS: By increasing the amounts of xanthan gum and transglutaminase, the viscosity of the samples was increased and syneresis was reduced significantly (P < 0.05). The kefir sample containing 150 ppm enzyme and 0.2% gum had the highest number of probiotic bacteria. Moreover, the highest organoleptic scores were found for this sample. CONCLUSIONS: It can be concluded that adding 150 ppm transglutaminase and 0.2% xanthan improved the vi- ability of probiotics and the physical and organoleptic characteristics of kefir.

Improving the stability of chitosan-gelatin-based hydrogels for cell delivery using transglutaminase and controlled release of doxycycline.

Although local cell delivery is an option to repair tissues, particularly using chitosan-based hydrogels, significant attrition of injected cells prior to engraftment has been a problem. To address this problem, we explored the possibility of stabilizing the chitosan-gelatin (CG) injectable hydrogels using (1) controlled release of doxycycline (DOX) to prevent premature degradation due to increased gelatinase activity (MMP-2 and MMP-9), and (2) transglutaminase (TG) to in situ cross-link gelatin to improve the mechanical stability. We prepared DOX-loaded PLGA nanoparticles, loaded into the CG hydrogels, measured DOX release for 5 days, and modeled using a single-compartmental assumption. Next, we assessed the influence of TG and DOX on hydrogel compression properties by incubating hydrogels for 7 days in PBS. We evaluated the effect of these changes on retention of fibroblasts and alterations in MMP-2/MMP-9 activity by seeding 500,000 fibroblasts for 5 days. These results showed that 90 % of DOX released from cross-linked CG hydrogels after 4 days, unlike CG hydrogels where 90 % of DOX was released within the first day. Addition of TG enhanced the CG hydrogel stability significantly. More than 60 % of seeded fibroblasts were recovered from the CG-TG hydrogels at day 5, unlike 40 % recovered from CG-hydrogels. Inhibition of MMP-2/MMP-9 were observed. In summary, controlled release of DOX from CG hydrogels cross-linked with TG shows a significant potential as a carrier for cell delivery.

Thermosensitive dendritic polyglycerol-based nanogels for cutaneous delivery of biomacromolecules.

Genetic skin diseases caused by mutations resulting in diminished protein synthesis could benefit from local substitution of the missing protein. Proteins, however, are excluded from topical applications due to their physicochemical properties. We prepared protein-loaded thermoresponsive poly(N-isopropylacrylamide)-polyglycerol-based nanogels exhibiting a thermal trigger point at 35°C, which is favorable for cutaneous applications due to the native thermal gradient of human skin. At≥35°C, the particle size (~200nm) was instantly reduced by 20% and 93% of the protein was released; no alterations of protein structure or activity were detected. Skin penetration experiments demonstrated efficient intraepidermal protein delivery particularly in barrier deficient skin, penetration of the nanogels themselves was not detected. The proof of concept was provided by transglutaminase 1-loaded nanogels which efficiently delivered the protein into transglutaminase 1-deficient skin models resulting in a restoration of skin barrier function. In conclusion, thermoresponsive nanogels are promising topical delivery systems for biomacromolecules. FROM THE CLINICAL EDITOR: Many skin disorders are characterized by an absence of a specific protein due to underlying gene mutation. In this article, the authors described the use of a thermoresponsive PNIPAM-dPG nanogel for cutaneous protein delivery in a gene knock-down model of human skin. The results may have implication for nano-based local delivery of therapeutic agents in skin.

A novel hemostatic sealant composed of gelatin, transglutaminase and thrombin effectively controls liver trauma-induced bleeding in dogs.

AIM: novel hemostatic sealant based on the in situ gel formation of gelatin catalyzed by thrombin and crosslinked by transglutaminase was designed. The aim of this study was to investigate the efficacy of the hemostatic sealant in control of bleeding caused by liver trauma in dogs. METHODS: Hepatic trauma that mimicked the grade III-IV rupture of liver was made in 20 dogs. The traumatic lesion was topically administered the hemostatic sealant (treatment group, n=10), or a thrombin solution (control group, n=10). The time to achieve hemostasis and the blood loss were measured. Contrast-enhanced ultrasound (CEUS) examination was performed in each animal on d 3, d 7, and d 10 d postoperatively to study the healing of the lesions. RESULTS: The mean time to achieve hemostasis in the treatment group was significantly shorter than that in the control group (1.20±0.33 vs 6.70±0.64 min, P<0.05). The mean blood loss in the treatment group was significantly less than that in the control group (47.22±8.61 vs 79.29±11.97 mL, P<0.05). In CEUS examination, the traumatic lesions in the treatment group became significantly smaller on d 3, and disappeared on d 7, whereas the lesions in the control group still existed on d 10. Ascites were never found during 10 d follow-up in the treatment group but were observed on d 3 and d 7 in the control group. CONCLUSION: Compared with thrombin, the novel hemostatic sealant shows much better efficacy in hemostasis and may promote wound healing in dog liver trauma.

The treatment of collagen fibrils by tissue transglutaminase to promote vascular smooth muscle cell contractile signaling.

The enzyme tissue transglutaminase 2 (TG2) appears to play an important role in several physiological processes such as wound healing, the progression of cancer and of vascular disease. Additionally, TG2 has been proposed as a means of stabilizing collagen extracellular matrix (ECM) scaffolds for tissue engineering applications. In this report, we examined the effect of TG2 treatment on the mechanical properties of the ECM, and associated cell responses. Using a model ECM of fibrillar collagen, we quantitatively examined vascular smooth muscle cell (vSMC) response to untreated, or TG2 treated collagen. We show that cells respond to TG2 treated collagen with increased spreading, an increase in contractile response as indicated by elevated F-actin polymerization and myosin light chain phosphorylation, and increased proliferation, without apparent changes in integrin specificity or matrix topography. Comparative atomic force microscopy loading studies indicate that TG2 treated fibrils are 3 times more resistant to shearing force from an AFM tip than untreated fibrils. The data suggest that TG2 treatment of collagen increases matrix mechanical stiffness, which apparently alters the contractile and proliferative response of vSMC.

The potential of mannosylated chitosan microspheres to target macrophage mannose receptors in an adjuvant-delivery system for intranasal immunization.

A vaccine delivery system based on mannosylated chitosan microspheres (MCMs) was studied in vitro and in vivo. Bordetella bronchiseptica antigens containing dermonecrotoxin (BBD) were loaded in MCMs or chitosan microspheres (CMs). Fluorescence confocal microscopy indicated that BBD-loaded MCMs (BBD-MCMs) bound with mannose receptors on murine macrophages (RAW264.7 cells). In vitro experiments using macrophages demonstrated that BBD-MCMs had more effective immune-stimulating activity than BBD-loaded CMs (BBD-CMs). Mice intranasally immunized with BBD-MCMs showed significantly higher BBD-specific IgA antibody responses in saliva and serum than mice immunized with BBD-CMs (p<0.05). After challenge with B. bronchiseptica via the nasal cavity, groups treated with BBD-MCMs or BBD-CMs showed similar patterns with a high survival rate even though there was no significant difference between those groups. These results suggested that mannose moieties in the MCMs enhanced immune-stimulating activities through mucosal delivery due to a specific interaction between mannose groups in the MCMs and mannose receptors on the macrophages.

Transglutaminases: a meeting point for wheat allergy, celiac disease, and food safety.

Wheat is the staple cereal in many countries and its uses in manufactured foods are ever growing due to the technological qualities of gluten proteins. Transglutaminases (TG) are ubiquitous enzymes with many functions. They are able to transform proteins by deamidation and/or transamidation. This last reaction can cross-link proteins together. Intestinal tissue TG has been shown to play an important role in two kinds of immune reactions to wheat: celiac disease and wheat-dependent exercise-induced anaphylaxis. In addition, new epitopes have been suspected in cases of anaphylaxis to wheat isolates, a food ingredient consisting mainly of deamidated gluten proteins. As a microbial TG is included in many food technological processes, its safe use should be checked. This assessment must cover not only the safety of the TG itself but also that of the deamidated/cross-linked proteins generated by this enzyme. This article aims at discussing the possible consequences of using TG in food industry in the light of today knowledge about immune reactions to wheat.

In vitro study of the immune stimulating activity of an atrophic [correction of athrophic] rhinitis vaccine associated to chitosan microspheres.

Chitosan microspheres (CMs) were prepared by an ionic gelation process with tripolyphosphate and characterized. Bordetella Bronchiseptica Dermonecrotoxin (BBD), a major virulence factor of a causative agent of atrophic rhinitis (AR), was loaded on to the CMs for nasal vaccination. BBD-loaded CMs were observed as aggregated shapes although unloaded CMs were observed as relatively spherical ones. The average particle size of the BBD-loaded CMs was 4.39 microm. The lower the molecular weight of chitosan and the higher the medium pH, the greater was the release of BBD from the BBD-loaded CMs in vitro due to weaker intermolecular interaction between chitosan and BBD. Tumor necrosis factor alpha and nitric oxide from RAW264.7 cells exposed to BBD-loaded CMs were gradually secreted with time, suggesting that released BBD from CMs had immune stimulating activity of AR vaccine in vitro.

Anticuerpos séricos de clase IgA anti-transglutaminasa tisular en el diagnóstico y seguimiento de la enfermedad celíaca. Resultados de un estudio multicéntrico internacional / The serum IgA class anti-tissue transglutaminase antibodies in the diagnosis and follow up of coeliac disease. Results of an international multi-centre study

Objetivos: La evaluación de la fiabilidad del test de anti-transglutaminasa tisular (anti-TGt) para el diagnóstico de la enfermedad celíaca se ha efectuado habitualmente con sistemas de enzimoinmunoanálisis (ELISA) ligeramente diferentes, en grupos de pacientes seleccionados y casi siempre pequeños. Los objetivos de este estudio son los siguientes: (1) evaluar la fiabilidad de los anticuerpos IgA anti-TGt en el diagnóstico de la enfermedad celíaca; y (2) establecer la sensibilidad y especificidad de un kit comercial para la determinación cuantitativa de anticuerpos anti-TGt.Diseño: Cada centro participante en este estudio multicéntrico internacional recogió muestras de suero de tres grupos de sujetos: (1) pacientes celíacos comprobados antes de iniciar dieta sin gluten; (2) pacientes celíacos en dieta sin gluten durante al menos 12 meses; (3) controles enfermos y sanos. Métodos: Se determinaron anticuerpos anti-TGt por duplicado mediante un kit de ELISA comercial (EutTG Eurospital, Trieste, Italia). Resultados: Se seleccionaron los siguientes casos y controles: (1) 399 sujetos con enfermedad celíaca activa; (2) 351 casos tratados de enfermedad celíaca; (3) 432 controles. La repetición centralizada de la prueba se realizó sobre: (1) grupo a: 176 pacientes con enfermedad celíaca activa (media de anti-TGt, 21 unidades arbitrarias; (2) grupo b: 172 casos tratados de enfermedad celíaca (media de anti-TGt, 5 AU); (3) grupo c: 206 controles (media anti-TGt, 3 AU). En celíacos activos, los anticuerpos anti-TGt mostraron un descenso progresivo con la edad significativo, mientras que en los controles se encontró una tendencia opuesta. En los pacientes con enfermedad celíaca activa, los anticuerpos anti-TGt estuvieron significativamente más elevados en celíacos con enteropatía de grado III que en los que tenían una lesión de grado II. En los celíacos tratados, los valores medios de anti-TGt eran significativamente menores en aquellos que seguían una dieta estricta sin gluten que en los que hacían transgresiones. La sensibilidad y especificidad de la técnica Eu-tTG fueron, respectivamente, 90 y 96 por ciento. Conclusión: Los resultados de este estudio muestran que el test comercial empleado para la determinación de anticuerpos anti-TGt es una herramienta reproducible y válida para el diagnóstico y seguimiento de la enfermedad celíaca. (AU)

Relación de los anticuerpos IgA anti-transglutaminasa tisular con la situación morfológica de la mucosa intestinal en niños con enfermedad celíaca / Relation of anti-tissue transglutaminase IgA antibodies with the morphology of the intestinal mucosa in children with celiac disease

Objetivo: Se ha investigado si la presencia de anticuerpos IgA antitransglutaminasa tisular es un indicador de la existencia de lesión de la mucosa intestinal, tanto en pacientes celíacos, en sus distintas fases evolutivas, como en controles de diverso tipo.Métodos: En 260 muestras de 214 pacientes, elegidos aleatoriamente, se han determinado, por método ELISA, anticuerpos IgA antitransglutaminasa tisular utilizando transglutaminasa de hígado de cobaya, así como anticuerpos IgA e IgG anti-gliadina. En un subgrupo de 109 muestras se determinaron, además, anticuerpos IgA antitransglutaminasa con transglutaminasa humana recombinante. Los resultados de todos ellos se han comparado con los de la biopsia intestinal tomada en los mismos días.Resultados: Con transglutaminasa de cobaya se ha obtenido una sensibilidad de 0,92 y una especificidad de 0,97, con relación a la biopsia intestinal, con una validez total máxima de 0,90 que corresponde a un coeficiente de probabilidad positivo de 34. El empleo de transglutaminasa humana recombinante no mejora estos resultados, pero se obtienen menos valores dudosos. Los anticuerpos antigliadina, tanto de clase IgA como de clase IgG sólo han mostrado una sensibilidad de 0,86 y 0,42 y una especificidad de 0,77 y 0,76, respectivamente.Conclusiones: La determinación de anticuerpos IgA anti-transglutaminasa tiene gran capacidad diagnóstica, reflejando el estado de la mucosa intestinal y mejorando sensiblemente la de otros marcadores séricos. No obstante, su discriminación no es absoluta, por lo que la BI debe seguir siendo el patrón-oro de referencia para el diagnóstico definitivo de la enfermedad celíaca (AU)

Tissue transglutaminase is expressed as a host response to tumor invasion and inhibits tumor growth.

A stable extracellular matrix (ECM) constitutes an important part of host response mechanism against tumor growth and invasion. Tissue transglutaminase (TG), a calcium-dependent enzyme, can cross-link all major ECM proteins to form a stable ECM, because these cross-links are resistant to proteolytic and mechanical damage. TG can also enhance stability and strength of the ECM by its ability to facilitate the activation of transforming growth factor-beta. We hypothesized that TG ECM-promoting abilities form an important part of the host response mechanism against tumor growth. Increased expression of TG was observed in the ECM of the host tumor interface of subcutaneously implanted rat mammary adenocarcinoma R3230 Ac. TG expression was also detected in the endothelial cells and macrophages. We also detected the cross-link product at the host tumor interface and within the tumor tissue, showing that TG was active. Western blots showed TG was degraded into three fragments of 55-, 50-, and 20-kDa forms. When recombinant wild-type TG was applied to R3230 Ac implanted in rat dorsal skin flap window chamber, it caused significant growth delay at day 7 compared with recombinant inactive TG controls. Collagen was detected in increased amounts in TG treated tumors, suggesting augmentation of production and stability of the ECM. We conclude that TG forms a distinct part of host response system against and acts to inhibit tumor growth.