Crosslinking of wheat dough proteins by glucose oxidase and the resulting effects on bread and croissants

The crosslinking of wheat flour proteins results in significant improvements in the functional properties of baked products. In this research, the enzyme glucose oxidase was investigated for its crosslinking effect on the dough proteins of bread and croissants. The macroscopic effects resulting from the addition of glucose oxidase to the dough formulation were compared to changes seen at the molecular level in individual protein fractions. Treatment with glucose oxidase produced slight improvements in crumb properties but no increase in product volume. At the molecular level, crosslinking occurred mainly in the water-soluble (albumin and globulin) fraction and was demonstrated to involve both disulfide and non-disulfide linkages. The SDS-soluble and -insoluble glutenins, which make up much of the gluten network, were crosslinked to a much lesser extent, with mainly non-disulfide linkages. These findings corroborate our theories on the relationship of wheat protein crosslinking and functional properties. Specifically, we conclude that (i) crosslinking of albumin and globulin proteins enhances crumb properties and (ii) changes in croissant volume require crosslinking of the glutenin fraction of wheat proteins.     

Pastry Lift and Croissant Volume as Affected by Microbial Transglutaminase

Microbial transglutaminase forms nondisulfide covalent crosslinks in proteins and is increasingly being used in foods. We have previously demonstrated beneficial effects of microbial transglutaminase during breadmaking, which are comparable to traditional oxidizing improvers, hypothesized to act via formation of disulfide crosslinks. Transglutaminase substantially improved the lift of puff pastry. It also had a dramatic effect on the volume of yeasted croissants made with both white flour and a blend of wholemeal and white flour. Furthermore, these effects were preserved after the pastry and croissant doughs had undergone frozen storage for periods of up to 90 d. Transglutaminase, therefore, offers a potential solution to the problem of pastry and croissant dough deterioration on frozen storage.     

Effects of processing on biochemical and rheological properties of wheat gluten proteins

Protein solubility increases during mixing at water absorption suitable for bread doughs. The changes that result from heat treatment may be much greater than the differences normally found among wheat varieties. Sheeting brings about a reduction in the gluten content and increase in gel protein content of doughs. This may be due to increase in temperature during sheeting as a result of work input to the dough resulting in protein denaturation. Drying at higher temperatures causes denaturation of pasta dough protein. In addition, it has been shown that high temperatures contribute to the formation of the protein network. During extrusion processing proteins are denatured and chemical bonds are weakened as result of heat and shear through the extruder.     

Effects of processing on biochemical and rheological properties of wheat gluten proteins.

Protein solubility increases during mixing at water absorption suitable for bread doughs. The changes that result from heat treatment may be much greater than the differences normally found among wheat varieties. Sheeting brings about a reduction in the gluten content and increase in gel protein content of doughs. This may be due to increase in temperature during sheeting as a result of work input to the dough resulting in protein denaturation. Drying at higher temperatures causes denaturation of pasta dough protein. In addition, it has been shown that high temperatures contribute to the formation of the protein network. During extrusion processing proteins are denatured and chemical bonds are weakened as result of heat and shear through the extruder.     

面包制作过程中戊聚糖酶的作用机理

研究了戊聚糖酶对面包品质的影响.加酶面包的体积明显增加,在适宜加酶量下,面包综合品质较佳.加酶面团中可溶性戊聚糖含量显著增加,但加酶过量可导致面团中水可提取的阿拉伯木聚糖(WEAX)降解程度加剧.未加酶面团较原料面粉中的可溶性戊聚糖含量略有增加,WEAX亦有部分降解,但作用程度远低于加酶样品.适量戊聚糖酶对面包品质的改良作用的关键在于产生大量的相对分子质量适宜的WEAX.     

The formation and properties of wheat flour doughs

Among the cereal flours, only wheat flour will form a viscoelastic dough when mixed with water. The viscoelasticity appears to be because the gluten proteins are water compatible and thus will swell and interact. The gluten protein's large molecular size and low charge density appear to allow them to interact by both hydrogen and hydrophobic bonds. Wheat flour doughs are also unique in their ability to retain gas. This property appears to result from a slow rate of gas diffusion in the dough. The third major unique property of wheat flour doughs is their ability to set in the oven during baking, and thereby to produce a rigid loaf of bread. Although not clearly understood, this appears to be a heat-induced crosslinking of the gluten proteins.     

Isolation and characterisation of wheat flour proteins. IV.—effects on wheat flour proteins of dough mixing and of oxidising agents

Proteins dispersible in 3M -urea increased from about 65 to 85% during mixing of dough in Farinograph. The increase was positively correlated with length of mixing. Protein dispersibility was highest in doughs mixed with five-fold excess potassium iodate, and was lowest in flours. Intermediate amounts of proteins were extracted from doughs mixed in air, in a nitrogen atmosphere, or in air with a five-fold excess of potassium bromate. Proteins, extracted with 3M -urea from doughs mixed to optimum consistency, were separated by fractionation on Sephadex G-100 columns into at least 6 fractions, constituting 3 distinct peaks. Distribution of proteins in the peaks was measured by Kjeldahl-N, biuret, and absorbance at 280 mμ methods. Mixing in air, but not in a nitrogen atmosphere, increased substantially the amount of proteins with a molecular weight above 150,000 and decreased the amount of low molecular-weight proteins. The effects of mixing bromated doughs were similar to mixing plain doughs in air, but the chemical modifications in bromated doughs were much more pronounced. No such changes were observed in iodate-treated doughs. Similar changes in distribution of protein fractions were observed when the gross protein extracts or Sephadex-fractionated proteins were studied by electrophoresis on starch gel in the presence of 3M -urea.     

Flour quality and disproportionation of bubbles in bread doughs

The bread making process transforms wheat flour doughs into highly porous breads. Bread has been shown (Wang, Austin and Bell, 2011) to be a single, open cell that is massively interconnected giving it a maze-like structure that encompasses the entire volume. The solid strands are also porous and contain closed cells. How the bubbles in dough mix partition into these open and closed cells in bread is not known. This study was undertaken to track changes in bubbles in doughs using 3-D X-ray microtomography techniques as doughs proofed and were baked. The mechanical properties of doughs were measured to establish how dough rheology impacted bubble growth. The doughs were made with ‘medium strong’ Canadian flour (CWRS) and ‘weak’ Australian flours (Wylk). Both doughs had similar protein amounts and strain-hardening characteristics; however the CWRS dough was more elastic. The scans identified formation of clusters of partially-coalesced bubbles from which one cluster grew to form a massively interconnected, single, closed cell in doughs as doughs proofed. Microscopy studies confirmed that the open cell in breads was made of partially-coalesced bubbles. Compared to the dough made with the Australian flours, the dough made from Canadian flour had a thicker dough layer separating bubbles, smaller size bubbles and a slower rate of formation of the continuous structure. This study highlights the critical role of dough elasticity and the disproportionation phenomena of bubble growth in controlling the quality of cell structures in dough and baked products.     

Impact of ‘oxidizing’ and ‘reducing’ buckwheat sourdoughs on brown rice and buckwheat batter and bread

The influence of reducing and oxidizing buckwheat sourdoughs on the rheological, protein, and bread properties of buckwheat and brown rice flour was investigated. Batters and breads prepared with chemically acidified doughs, fresh pre-doughs, and fresh pre-doughs containing glutathione (3 mM) were used as controls. No significant differences were observed after the addition of reducing and oxidizing sourdoughs in all trials. Proteolysis was observed after proofing time in buckwheat and brown rice batters, respectively. Acidified doughs increased the elasticity and the gelatinization temperature of buckwheat batters. No notable microstructure changes were detected in brown rice batters. The extension of fermentation time in sourdough caused a slight decrease in bread volumes in all trials. Sourdoughs increased the bread volume and decreased the crumb hardness of buckwheat breads. In trials with brown rice flour, the addition of sourdough did not show relevant volume differences as compared to the controls, except big voids in sourdough bread crumb. Linear correlations between hardness, volume, and cells’ density were observed. However, no clear correlations among rheological parameters and bread characteristics could be detected. These results indicated that the applied strains were responsible for the leavening capacity of the yeast during the proofing time and for crumb structure in trials with buckwheat and brown rice flour. Applied sourdoughs were able to change the molecular, and bread properties of buckwheat and brown rice bread.     

Effect of durum wheat varieties on bread quality

In this study, the effect of different durum wheat varieties on the sensorial and nutritional quality of bread was assessed. In particular, bread manufactured with six wheat cultivars was compared with bread based on commercial semolina mixtures. X‐ray microtomography was used to characterise the final cellular structure of the baked bread. The textural properties of baked bread and doughs were also investigated. In addition, the glucose equivalent (GE) of bread was also evaluated. Finally, the microstructural, sensorial and textural parameters were correlated with each other. Results highlighted that Cappelli and Saragolla bread samples showed higher protein and total soluble fibre content, respectively, in comparison with the reference samples. Moreover, the GE of the Cappelli, Claudio and Saragolla bread samples was similar to that of the reference samples and lower than that of the other samples. However, the sensorial properties of all bread samples showed a very positive score (up to 7.7).     

Structural and farinographic changes during mixing of a yeast sweet dough

Sweet dough requires longer mixing time than salty or white pan bread doughs to reach a developed stage. Although many studies have dealt with the effect of mixing time on dough, few have referred to yeast sweet doughs. The aim of this study was to evaluate if the changes in dough microstructure during different stages of mixing were the same between sweet pan bread and white pan bread, using as control a flour water system. Scanning electronic microscopy (SEM) and some bread characteristics were used as evaluation parameters. Doughs were prepared in a Brabender Farinograph instrument. Different mixing times were used for each formulation, which correspond to common farinographic parameters such as: arrival time, peak time, departure time, etc. Farinographic consistency was evaluated at those times. Results showed that sweet dough farinogram was quite different from those obtained from the other two samples; it starts with a low consistency value (260 BU), and after 8 min of mixing it began to increase until almost reaching the 500 BU line; then the graphic follows the classical curve. Larger products were obtained from arrival time to departure time for both formulations. SEM showed that as mixing proceeds the dough structure opens, changing its appearance from a compact structure at the beginning to a very open one at the end of the mixing process. SEM also showed that the process of mixing is the same for the two samples and control; however, the time needed to reach each stage was different among samples. Farinogram can be used to get information about mixing behavior of yeast sweet doughs. The study of mixing can be easier using sweet dough formulations because it was possible to get more points between the onset of mixing and full dough development, and the process was very similar no matter the formulation.     

Rebuilding gluten network of damaged wheat by means of glucose oxidase treatment

The disrupted gluten structure of infested wheat flours leads to low‐quality doughs unusable in bread‐making processes. Enzymes are replacing chemical treatments in the food industry as a tool to treat weak flours. Glucose oxidase is one of the most promising oxidative enzymes, although its efficiency compared with the alcohol‐soluble fraction of gluten proteins has not yet been demonstrated. If this enzyme could restore the broken covalent bonds between glutenin subunits, the gluten network of damaged wheat flour would recover its native structure and functionality. This treatment would allow bakers to use damaged flour, reducing the economic losses caused by this plague around Europe and North Africa. Electrophoretic studies demonstrated the formation of high‐molecular‐weight aggregates in the glutenin fraction, which had a characteristic thermal stability depending on enzyme dosage. Those molecular studies agreed with the bread‐making assays made with maximum enzyme dosage and microstructure determination. Overall results showed that glucose oxidase is a real alternative to traditionally used chemical oxidants. It acted specifically on the high‐molecular‐weight glutenin subunits of damaged wheat, forming dityrosine crosslinks between the wheat proteins, which reinforced the gluten network and gave away the dough functionality. Copyright © 2007 Society of Chemical Industry     

Biotechnology of Wheat Quality

Wheat gluten proteins are largely responsible for the visco-elastic properties that allow doughs to be processed into bread and various other food products including cakes, biscuits (cookies), pasta and noodles. Detailed biochemical and biophysical studies are revealing details of the molecular structures and interactions of the individual gluten proteins, and their roles in determining the functional properties of gluten. In particular, one group of gluten proteins, the high molecular weight (HMW) subunits of glutenin, have been studied in detail because of their role in determining the strength (elasticity) of doughs. The development of robust transformation systems for bread wheat is now allowing the role of the HMW subunits to be explored experimentally, by manipulating their amount and composition in transgenic plants. Such studies should lead to improvement of the processing properties of wheat for traditional end uses and the development of novel end uses in food processing or as raw material for other industries. © 1997 SCI.     

Water Content, Water Soluble Fraction, and Mixing Affect Fundamental Rheological Properties of Wheat Flour Doughs

Fundamental rheological behavior of wheat flour doughs were compared with empirical results from a farinograph. Water contents of 42-47% and mixing times of 8, 16, and 24 min were examined. The storage modulus, G′, decreased when water content was increased from 42 to 45%. Dough produced from water extracted flours had high G′ and low δ. Increasing water content or mixing time decreased G′. There was a linear relationship (slope = 0.30) between G′ and the farinogram peak height for the reconstituted (water-soluble fraction added back), flour-water, and freeze-dried flour-water doughs. The extracted flour doughs also showed a linear relationship between G′ and farinogram peak height with slope = 1.07.     

Effects of two barley β-glucan isolates on wheat flour dough and bread properties

The effects of wheat flour fortification with two different molecular weight barley β-glucan isolates (1.00 × 10⁵, BG-100 and 2.03 × 10⁵, BG-200) on the rheological properties of dough and bread characteristics, using flours from two wheat cultivars that differ in their breadmaking quality, have been examined. The farinograph water absorption of doughs and the moisture content and water activity of the breads increased with increasing β-glucan content; the β-glucan isolate with the higher molecular weight (BG-200) exerted a greater effect than did BG-100. The addition of β-glucans to the dough formula increased the development time, the stability, the resistance to deformation and the extensibility of the poor breadmaking quality doughs, as well as the specific volumes of the respective breads, exceeding even that of the good breadmaking cultivar. Furthermore, the colour of the bread crumbs got darker and their structure became coarser, whereas the bread crumb firmness decreased with increasing level of β-glucan addition. Generally, the BG-200 was more effective in increasing the specific bread volume and reducing the crumb firmness, especially when used to fortify the poor breadmaking quality flour. The results further indicate a requirement for optimisation of the fortified doughs (level and molecular size of the β-glucan) to maximise bread quality attributes (loaf volume, texture, and staling events).     

Scanning electron microscopic observations of dough and bread supplemented with Gastrodia elata Blume powder

Dough and bread prepared from wheat flour containing varying amounts of added Gastrodia elata Blume (GEB) rhizome powder [0, 0.5, 1.0, 1.5, and 2.0% (w/w)] were examined by scanning electron microscopy (SEM) during fermentation and baking. The structure of the doughs containing added GEB was found to be related to the protein matrix. Further, it was found that large starch granules and strings of small starch granules play an important role in dough structure. The control dough (no added GEB) had a membrane-like structure, and doughs with 0.5–1.0% added GEB had membrane-like structures that were more developed than those of the control, resulting in increased bread volumes. At 1.5–2.0% GEB levels, however, the doughs tended to have mesh-like structures and result in decreased bread volumes. The dough samples with 0.5 and 1.0% added GEB powder had well-developed gluten matrices with evenly dispersed starch granules. These samples resulted in breads with numerous gas bubble eruptions on their surfaces and consequently in larger loaf volumes than were obtained at other levels of GEB. After the second fermentation, many expanded starch granules were observed and these starch granules were dispersed more evenly than after the first fermentation. In 0.5–1.0% GEB bread, many of the large starch granules had expanded after fermentation, but small starch granules had not. The data obtained in this study suggest that bread baked with 0.5–1.0% GEB exhibits a better loaf volume due to the more complete development of a gluten matrix.     

Suitability of various australian wheats for chinese-style steamed bread

Three methods of preparation of steamed bread on a laboratory scale were evaluated to determine the one most suitable for discriminating between flour samples. The preferred method was then applied to a number of wheat varieties at different protein levels to select the most suitable ones for this product, and to determine the key analytical parameters of the most suitable flour. Flour colour and protein content appeared to be more important than other physical and chemical properties. Flour samples with high protein content (greater than 12%) and strong doughs gave bread with a wrinkled surface and dark colour, whereas low protein (less than 10%) soft wheat flours gave poor texture and eating quality, although the surface was smooth. It is suggested that flours of medium protein content (10–12%) and medium dough strength are the most suitable for steamed bread. Processing conditions affected steamed bread quality. Blending of high protein hard strong wheats with soft weak wheat could be practical for preparing flour for steamed bread.     

Rheological,microstructural and sensorial properties of durum wheat bread as affected by dough water content

In this paper the influence of water content on the rheological, microstructural and sensorial properties of durum wheat bread was evaluated. In order to evaluate bread quality, oscillation measurements, stress relaxation test and creep–recovery measurements were performed on dough samples, whereas tomographic and sensorial analyses were performed on baked bread samples. Results of the rheological analysis highlighted that both the storage and loss moduli (G′, G″) showed a descending trend with the increase of the water content. This is also confirmed by stress relaxation tests. Creep–recovery tests for strong doughs (with low water content), recorded greater resistance to deformation, therefore a smaller creep strain than the softer doughs. These results were reflected in the microstructural properties of the bread; an increase in water content caused an increase in the percentage volume of pores. Regarding the sensorial properties, the overall acceptability of the investigated bread samples was low for both the lowest and the highest water contents, and this was due primarily to the compact crumb with small bubbles and high crust firmness for the former and to the loaf volume collapsed with irregular distribution of very large bubbles for the latter. Therefore, the bread samples with intermediate water content were preferred by the panelists.     

Glucose oxidase effect on dough rheology and bread quality: A study from macroscopic to molecular level

Enzymes are used in baking to improve dough handling properties and the quality of baked products. Glucose oxidase (GO) is an enzyme with oxidizing effect due to the hydrogen peroxide released from its catalytic reaction. In this study, the macroscopic effect of increasing glucose oxidase concentrations on wheat dough rheology, fresh bread characteristics and its shelf life during storage was determined. A reinforcement or strengthening of wheat dough and an improvement of bread quality can be obtained with the addition of GO, although inverse effects were obtained when excessive enzyme levels were added. The analysis of the gluten proteins at molecular level by high performance capillary electrophoresis and at supramolecular level by cryo-scanning electron microscopy revealed that the GO treatment modified gluten proteins (gliadins and glutenins) through the formation of disulfide and non-disulfide crosslinks. The high molecular weight glutenin subunits showed to be the most susceptible glutenin fraction to the oxidation action of GO. Excessive addition of GO produced an excessive crosslinking in the gluten network, responsible of the negative effect on the breadmaking properties.     

Assessing Influence of Protein Source on Characteristics of Gluten-Free Breads Optimising their Hydration Level

Most gluten-free products have lower protein content than their counterparts with wheat flour. The addition of exogenous proteins could not only be a good option to compensate for this reduction but also a tool to create gluten-free products rich in protein. However, the different water-binding capacities of proteins modify dough rheology, which also affects bread volume. Therefore, this study aimed to analyse the incorporation of a high percentage (30%) of several proteins (rice, pea, egg white and whey protein) in gluten-free breads whose hydration levels were adjusted for each protein to achieve the maximum volume. In this way, the breads with vegetal proteins required a higher amount of water than the breads with animal proteins. Moreover, all enriched breads exhibited lower maximum volume values than control, and the ones with whey protein presented the lowest volumes overall. From these results, the rheological behaviour and characteristics (colour, texture and weight loss) of optimised doughs and breads were measured. The doughs with whey protein presented the highest G′ and G″ values due to their low hydration level, and the ones with egg white protein were very watery. Regarding colour, the addition of protein led to darker crusts, with the ones with whey protein being the darkest. With respect to the control, breads with animal proteins exhibited higher hardness, especially with whey protein, while the ones with vegetal proteins did not present significant differences.