Microbial Transglutaminase: General Characteristics and Performance in Food Processing Technology

The enzyme microbial transglutaminase (MTGase) (EC 2.3.2.13) catalyzes acyl transfer reactions, deamidation, and inter and intramolecular crosslinks between amino acid residues of glutamine and lysine. Its application in foods alters the technological properties of food proteins, such as their emulsifying capacity, gelation, viscosity, water holding capacity, and foam formation and stability. The commercial application of the enzyme in Brazil is limited almost entirely to the restructured meat industry and dairy beverages, but several studies have demonstrated its potential for commercial application in vegetable products such as soy, which is little exploited in the market. The application of this enzyme to vegetable derivatives allows for the development of new products or improvement of the functional properties of traditional foods. The objective of this article is to offer a comprehensive overview of the general characteristics and function of the enzyme microbial transglutaminase, with emphasis on its application in foods.     

Processing conditions and transglutaminase sources to “drive” the wheat gluten dough quality

Gluten proteins are highly impacting the quality of various gluten-based products, and transglutaminases (TGs) are used to influence the protein cross-linking. In this study we monitored the interplay of “harsh” and “mild” gluten processing for dough mixing and pasta-like sheet production and TGs from a commercial and newly sourced bacteria (SB6). Despite the harshly separated gluten presenting strongly cross-linked proteins in the beginning of the mixing, similar levels of polymerization were achieved at the optimum mixing time but with differences in the secondary protein structure. TG addition increased polymerization in wheat doughs, possibly as a result of increased glutenin polymerization, while gliadins become more soluble with SB6. This enzyme also dramatically increased polymerization in mild gluten. These results show that an adequate investigation when using TGs and gluten from various origins is necessary to adequately predict the quality in various gluten-based products, thus, of great relevance to the food industry.Industrial relevanceCurrently, there is a mounting trend towards the modification of gluten proteins to improve technological features and functionality. In breadmaking, when weak flour (low protein content) is used or general stabilization is desired for technological purposes, additives can be used to stabilize the gluten protein matrix. The use of transglutaminase (TG) has grown in popularity as they promote specific cross-linking between residues of glutamine and lysine in proteins. Another way of improving dough functionality is by increasing the oxidation of disulfide groups by adding gluten which is a co-product of the starch industry. Industrial production of gluten includes the use of heating and shear forces, which may impact gluten dough-forming ability. Thus, increased understanding of the interplay of gluten processing and the impact of choice of the TG origin in gluten dough quality is highly applicable in food industry.     

Use of dairy proteins and microbial transglutaminase to obtain low-salt fish products from filleting waste from silver carp (Hypophthalmichthys molitrix)

Restructured fish products from the filleting waste from silver carp (Hypophthalmichthys molitrix) were obtained using sodium caseinate (1%), whey protein concentrate (WPC) (1%) or MTGase (0.3%) at different levels of salt (0%, 1% or 2%). The restructured products were obtained by incubating at 40 °C for 60 min and then at 90 °C for 20 min. Changes in the mechanical properties were studied by measuring changes in the textural profile analyses, punch test and torsional test analyses and expressible water. The mechanical properties increased when the salt level was also increased. The mechanical properties of non-salted and low-salt products were increased by using dairy proteins. Sodium caseinate had a greater effect in improving mechanical properties than WPC. MTGase increased expressible water. The results obtained showed that the mechanical properties of low-salt restructured fish products can be improved by using dairy proteins combined with MTGase, with a slight increase in expressible water.     

TRANSGLUTAMINASE IN DAIRY PRODUCTS: CHEMISTRY, PHYSICS, APPLICATIONS

Literature on the effects of microbial transglutaminase on various dairy‐based systems is discussed. Beginning with a short synopsis on the development of microbial transglutaminase as a functional tool for modifying foods, the principles of reactions catalyzed by transglutaminase and their structural implications, as well as the mechanisms of formation and cleavage of isopeptide bonds are reviewed. After summarizing the present knowledge on the specificity of microbial transglutaminase towards milk proteins, including reactions determined by individual lysine and glutamine residues, emphasis is placed on the effects of enzymatic cross‐linking on physicochemical properties in foods and, particularly, dairy‐based systems. Discussed are implications of cross‐linking on acidified milk gels including yogurt and effects on single milk protein fractions, with respect to several physicochemical properties including rheology and mechanical properties of these systems, but also syneresis, and emulsification behaviour.     

乳和乳制品中抗生素残留的危害及检测方法

综述了乳与乳制品中抗生素残留现状和危害，比较了抗生素检测Delvotest法与其他几种方法。认为Delvotest法优于其他几种方法，建议有关部门尽快引进该技术，以加强我国乳和乳制品中的抗生素监测，解决目前乳业中面临的难题。     

Texturisation and modification of vegetable proteins for food applications using microbial transglutaminase

Microbial transglutaminase (MTG) isolated from Streptomyces mobaraensis has been available on a commercial scale for several years. MTG generates inter- and intramolecular cross-links between γ-carboxylamide groups of glutamine residues and ɛ-amino groups of lysine residues in proteins. Due to its great potential to improve various functional properties of proteins, MTG is mainly used to enhance texture, stability, and water binding. Application of MTG for the production of plant protein-based foodstuffs such as tofu, noodles, bread and bakery products, is still limited to raw materials from soybean and wheat. However, with the increasing demand for vegetarian foods, the utilisation of novel proteins as functional ingredients, e.g. from peas, lupins, sesame, and sunflower, seems promising. To open new horizons for MTG application, this review aims at demonstrating the actual potential of MTG in processing foodstuffs based on vegetable proteins. Particular focus was laid on novel plant protein sources suitable for cross-linking with MTG. Furthermore, strategies for improving texture and nutritive value of the proteins are discussed.     

Liver protein expression in dairy cows with high liver triglycerides in early lactation

Fatty liver is a frequent subclinical health disorder in dairy cows that may lead to disorders related to the liver function. However, the effect of triglyceride (TG) accumulation on liver metabolic pathways is still unclear. The objective was, therefore, to characterize quantitative differences in the liver proteome between early lactation dairy cows with a low or high liver TG content. The liver proteome analysis indicated that a high liver TG content in early lactation dairy cows is associated with increased oxidation of saturated fatty acids, oxidative stress, and urea synthesis and decreased oxidation of unsaturated fatty acids. Furthermore, liver gluconeogenesis is apparently not impaired by an increased liver TG content. Based on correlations between liver proteins and plasma components, we suggest that future studies investigate the sensitivity and specificity of plasma aspartate aminotransferase, β-hydroxybutyrate, total bilirubin, total bile acids, and γ-glutamyltransferase for potential use as blood-based biomarkers for early detection of fatty liver in dairy cows. Our study is the first to study the proteome of dairy cows with naturally occurring fatty liver in early lactation.     

Effect of enzymatic cross-linking of milk proteins on functional properties of set-style yoghurt

This paper presents research on the effect of enzymatic cross-linking of milk proteins on the properties of yoghurt. Whole milk was incubated with transglutaminase (TG) prior to fermentation (2 h, 40°C, E/S ratio 1/2000). Enzyme action was stopped by heating (1 min, 80°C). Skim-milk was treated by simultaneous use of TG and thermophilic yoghurt starter culture without inactivation of the enzyme. A TG treatment of milk prior to fermentation led to prolonged fermentation, while the concomitant use of TG and culture had no influence on fermentation time. Post acidification of yoghurt during storage was lower for products from enzyme-treated milk. This applies both for products cross-linked prior to fermentation with enzyme inactivation, and for simultaneous use of culture and TG without inactivation of the enzyme. Scanning electron microscopic studies revealed that a TG treatment of milk led to reduced mesh sizes of the protein network, and a more regular distribution of the proteins in the yoghurt gel. As a result, yoghurt products from enzyme-treated milk showed increased gel strength and less syneresis, especially when the enzyme was not inactivated. Sensory studies revealed that odour and consistency properties of products from TG-treated milk were assessed as less 'yoghurt specific'. On the other hand, products from enzyme-treated milk were described as being more creamy, indicating that a TG treatment may simulate fat in fermented milk products.     

Addition of crude tiger nut protein extract affects stiffness of enzymatically cross‐linked dairy proteins

The influence of crude tiger nut protein extract on the gel properties of enzymatically cross‐linked dairy proteins was investigated. Enzymatic cross‐linking of dairy proteins in the presence of crude tiger nut proteins caused the formation of larger casein polymers and increased the degree of polymerisation. Gel stiffness of acidified products containing whey proteins was higher when cross‐linking occurred in the presence of crude tiger nut proteins. The results are relevant for improving the textural characteristics of acidified aqueous tiger nut extract (tiger nut milk) enriched in dairy proteins.     

乳及乳制品中残留青霉噻唑酸钾的检测方法

通过分光光度法检测乳及乳制品中青霉素钾的主要酶解产物青霉噻唑酸钾,间接检测了乳及乳制品中是否添加过β-内酰胺酶。通过实验该方法的工作曲线相关系数为0.9997,检测限为2.5 mg/L,回收率在93.2%～106.3%之间,RSD为2.9%～4.5%。在30组样品中应用效果良好,表明该方法简单、快速准确、灵敏度高,适用于乳及乳制品中青霉噻唑酸钾残留量的检测。     

高静压技术在乳及乳制品中的应用进展

现在人们热衷于开发更高安全性、营养质量和具有独特物化特性的新型乳制品。高静压技术对产品赋予特殊的功能性质的潜力巨大 ,从而有助于开发更高营养和更好感观质量的新型乳制品 ,形成新的组织形态 ,并提高了产品货价期 ,所以在乳及乳制品中的应用逐渐增加。这篇文章总结了高静压技术在乳及乳制品中的应用进展 ,并就压力的潜在作用———导致乳制品的修饰改性 ,深入探讨了压力处理条件下乳脂肪和乳蛋白质的变化。     

Bioactive peptides: Production and functionality

Milk proteins exert a wide range of nutritional, functional and biological activities. Many milk proteins possess specific biological properties that make these components potential ingredients of health-promoting foods. Increasing attention is being focused on physiologically active peptides derived from milk proteins. These peptides are inactive within the sequence of the parent protein molecule and can be liberated by (1) gastrointestinal digestion of milk, (2) fermentation of milk with proteolytic starter cultures or (3) hydrolysis by proteolytic enzymes. Milk protein derived peptides have been shown in vivo to exert various activities affecting, e.g., the digestive, cardiovascular, immune and nervous systems. Studies have identified a great number of peptide sequences with specific bioactivities in the major milk proteins and also the conditions for their release have been determined. Industrial-scale technologies suitable for the commercial production of bioactive milk peptides have been developed and launched recently. These technologies are based on novel membrane separation and ion exchange chromatographic methods being employed by the emerging dairy ingredient industry. A variety of naturally formed bioactive peptides have been found in fermented dairy products, such as yoghurt, sour milk and cheese. The health benefits attributed to peptides in these traditional products have, so far, not been established, however. On the other hand, there are already a few commercial dairy products supplemented with milk protein-derived bioactive peptides whose health benefits have been documented in clinical human studies. It is envisaged that this trend will expand as more knowledge is gained about the multifunctional properties and physiological functions of milk peptides.     

Enzymatic crosslinking and food allergenicity: A comprehensive review

Food allergy has become a major global public health concern. In the past decades, enzymatic crosslinking technique has been employed to mitigate the immunoreactivity of food allergens. It is an emerging non-thermal technique that can serve as a great alternative to conventional food processing approaches in developing hypoallergenic food products, owing to their benefits of high specificity and selectivity. Enzymatic crosslinking via tyrosinase (TYR), laccase (LAC), peroxidase (PO), and transglutaminase (TG) modifies the structural and biochemical properties of food allergens that subsequently cause denaturation and masking of the antigenic epitopes. LAC, TYR, and PO catalyze the oxidation of tyrosine side chains to initiate protein crosslinking, while TG initiates isopeptide bonding between lysine and glutamine residues. Enzymatic treatment produces a high molecular weight crosslinked polymer with reduced immunoreactivity and IgE-binding potential. Crosslinked allergens further inhibit mast cell degranulation due to the lower immunostimulatory potential that assists in the equilibration of T-helper (Th)1/Th2 immunobalance. This review provides an updated overview of the studies carried out in the last decade on the potential application of enzymatic crosslinking for mitigating food allergenicity that can be of importance in the context of developing hypoallergenic/non-allergenic food products.     

Effects of Transglutaminase on SDS-PAGE Patterns of Wheat, Soy, and Barley Proteins and their Blends

Transglutaminase (TG) catalyzes the formation of nondisulfide covalent crosslinks between pep‐tide‐bound glutaminyl residues and ε‐amino groups of lysine residues in proteins. TG can be used for polymerizing proteins from 1 or more sources through formation of intermolecular crosslinks. This study investigated, by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, polymers created by the action of TG on proteins of wheat, soy, barley, wheat‐soy, and wheat‐barley blends. Electrophoretic results showed that with increasing incubation time, the crosslinking reaction is substantially increased, with progressive decrease or disappearance of some protein monomers. Densitometric results showed that soy proteins were the best substrates of TG while barley and wheat proteins were similar in reactivity.     

食品中乳蛋白的重要作用

乳蛋白具有许多令人关注的特性，包括营养、理化、功能、工艺、生理保健等特性．被广泛地应用于乳制品、乳饮料和保健食品等．作者首先介绍了乳蛋白的一些特性，然后描述了富含蛋白质的主要乳制品，包括产量较大的干酪、乳粉和产量较小的特殊制品如酪蛋白和乳清蛋白制品．     

Monitoring the authenticity of low-fat yogurts by an artificial neural network

The growing consumption of low- and reduced-fat dairy products demands routine control of their authenticity by health agencies. The usual analyses of fat in dairy products are very simple laboratory methods; however, they require manipulation and use of reagents of a corrosive nature, such as sulfuric acid, to break the chemical bounds between fat and proteins. Additionally, they generate chemical residues that require an appropriate destination. In this work, the use of an artificial neural network based on simple instrumental analyses, such as pH, color, and hardness (inputs) is proposed for the classification of commercial yogurts in the low- and reduced-fat categories (outputs). A total of 108 strawberry-flavored yogurts (48 probiotic low-fat, 36 low-fat, and 24 full-fat yogurts) belonging to several commercial brands and from different batches were used in this research. The statistical analysis showed different features for each yogurt category; thus, a database was built and a neural model was trained with the Levenberg-Marquardt algorithm by using the neural network toolbox of the software MATLAB 7.0.1. Validation with unseen data pairs showed that the proposed model was 100% efficient. Because the instrumental analyses do not require any sample preparation and do not produce any chemical residues, the proposed procedure is a fast and interesting approach to monitoring the authenticity of these products.     

钙与乳蛋白的相互作用机制及对其功能特性的影响研究进展

钙作为人体骨骼生长和调节生理功能的重要物质,广泛存在于乳品中。钙与乳中蛋白质如αs-酪蛋白、β-酪蛋白、κ-酪蛋白、α-乳白蛋白和β-乳球蛋白等发生相互作用,影响乳蛋白的结构稳定性,进而使其热稳定性、乳化性、溶解性、起泡性和凝胶性发生改变。本文综述了钙与各种乳蛋白之间的相互作用机制以及对其功能特性的影响,对系统理解钙与乳蛋白的结合具有重要意义,为钙类功能性乳品的深入研究以及工业化生产提供理论依据。     

Educational interventions to address misconceptions about antibiotic residues in milk can alter consumer perceptions and may affect purchasing habits

The industrialization of the agri-food industry and resultant decrease in the number of people employed on farms has contributed to a knowledge gap among consumers about food production processes. A commonly reported concern of dairy consumers is the use of antibiotics in dairy animals, even though these drugs are an important tool for promoting animal health and welfare and food safety. The extent to which consumers are aware of antibiotic residue avoidance practices in dairy production is unknown, and it is unclear whether acquisition of such knowledge could affect purchasing behavior and perceptions of dairy farming. The objectives of this study were to assess consumers' perceptions about the quality and production of dairy products in the United States and determine whether educational materials on processes that limit the occurrence of antibiotic residues in milk can change consumers' perceptions of dairy products and purchasing behaviors. We surveyed 804 consumers and assigned them to 1 of 3 interventions: (1) a control arm (reading the content of the Dairy page of the USDA's myplate.gov website); (2) an educational brochure on the processes that prevent antibiotic residues in milk; and (3) a video on the same processes. We found that a majority (86.1%) of participants believe that the quality of dairy products in the United States is high, although many had concerns about the treatment of dairy animals and chemicals (pesticides, antibiotics, hormones) in dairy products. Compared with the control intervention, the brochure was associated with a significant decrease in the level of concern consumers had about chemicals in their milk [−0.20 points on a Likert scale, 95% confidence interval (CI), −0.32 to −0.08] and a significantly increased comfort in purchasing conventional dairy products (odds ratio 2.43, 95% CI, 1.62 to 3.66). The video was associated with even stronger effects: a 0.29-unit decrease in the level of concern about chemicals in milk (95% CI, −0.42 to −0.016) and 2.94 times greater odds of purchasing conventional dairy products (95%, CI 1.92 to 4.49). Although consumer food decision making is complex and driven by multiple factors, it appears that education about the processes that promote food safety can reassure consumers about their concerns and potentially affect purchasing habits.     

乳蛋白作为食品配料的价值与应用

通过对乳蛋白质体系的组成、结构、营养和物理化学性质的分析，阐述乳蛋白的价值以及富含优质蛋白的乳制品作为蛋白性配料的应用。同时介绍一些特殊乳蛋白如乳铁蛋白、免疫球蛋白等的市场前景。