A novel approach for improving yeast viability and baking quality of frozen dough by adding biogenic ice nucleators from Erwinia herbicola

Freezing deteriorates the baking quality of frozen bread dough by causing lethal injury to yeast cells and depolymerization to the gluten network. To investigate the potential of biogenic ice nucleators in frozen food applications, the effect of extracellular ice nucleators (ECINs) from Erwinia herbicola on the baking quality of frozen dough upon three freeze/thaw cycles were investigated. With addition of ECINs to the activity of 2.4 × 10⁶ units per gram of dough, hardening of bread crumb caused by three freeze/thaw cycles was alleviated by about 50% compared to the control. Additionally, the bread from frozen dough with added ECINs showed 50% larger specific volume compared to the control. The mechanism of cryoprotective effects from ECINs was possibly that ECINs helped in preserving the viability of yeast cells during freeze/thaw cycles. ECINs were able to improve the viability of log-phase and stationary-phase yeast cells in suspensions by about 100 and 10 fold, respectively, and viability of yeast in the frozen dough by 17%. This study revealed the potential of ECINs as a cryoprotectant for applications in the food and biotechnology industries.     

Soy ingredients stabilize bread dough during frozen storage

Bread with 48.5% soy ingredients was assessed for quality during frozen storage of the dough. Soy protein was hypothesized to prevent water migration during frozen storage, thereby producing dough that would exhibit fewer structural changes than traditional wheat bread. Wheat and soy bread were baked from dough that was fresh or frozen (−20 °C, 2 or 4 wks). Dough and bread were assessed for physical properties including moisture content, percent “freezable” and “unfreezable” water, dough extensibility, and bread texture. The bread was subjected to an untrained sensory panel. The soy bread was denser, chewier, and had a higher moisture content than wheat bread. When baked from fresh or frozen dough, soy bread was rated “moderately acceptable” or higher by 70% of panelists. Soy minimized changes in dough extensibility and resistive force to extension, leading to minimal changes in bread hardness. Although consumers could distinguish between bread baked from soy dough that was fresh or frozen for 4 wks, sensorial and textural data suggested that the rate at which the quality of the soy dough deteriorated was slower than that of wheat dough. In conclusion, the dough of consumer-acceptable soy bread retained quality characteristics during frozen storage slightly better than wheat dough.     

Effects of glycerol on water properties and steaming performance of prefermented frozen dough

The amount of ice in both unfrozen steamed bread dough (UFD) and prefermented frozen steamed bread dough (PFD) with and without glycerol was investigated by differential scanning calorimetry (DSC). The quality of unfrozen steamed bread (UFB)/prefermented frozen dough steamed bread (PFB) was also evaluated. Frozen stability and steaming performance of prefermented frozen dough were negatively correlated with ice crystal growth. Glycerol effectively prevented the formation of ice crystals during freezing and frozen storage, maintaining the quality of steamed bread from prefermented frozen dough even over a period of 30 days. The best steamed bread performance was observed with the dough containing 2% of glycerol (flour weight basis) addition. Prefermenting conditions significantly affected the quality of UFB/PFB. The highest quality scores of steamed bread from prefermented frozen dough were obtained from 32 °C and 85% rh for 40 min.     

Staling of fresh and frozen gluten-free bread

The difficulty in finding gluten-free bread and its high price make it necessary to prolong its shelf life to facilitate its availability. Freezing is an interesting alternative. The storage of bread at over zero temperatures, 20 °C and 4 °C, showed faster staling at refrigerator temperatures. A good relationship between crumb firmness and the extent of starch recrystallization was obtained, although the effect of water loss was also detected. The study of freezing and frozen storage at −14 °C and −28 °C for 7 days showed a substantial effect of the storage temperature on gluten-free bread quality and shelf life. Breads stored at −28 °C retained a quality similar to that of fresh breads while a marked deterioration of the breads stored at −14 °C was observed. This effect, the strongest on bread texture, was a result of starch recrystallization. The glass transition, T_g’ and onset of ice melting, T_m’ of the maximally freeze-concentrated bread crumb were −37.1 ± 0.6 °C and −19.3 ± 0.2 °C respectively. The higher amount of unfrozen water at −14 °C could explain the acceleration of reactions responsible for bread staling during frozen storage. The use of storage temperatures below T_m’ is recommended to retain high quality of the gluten-free bread during frozen storage.     

Impact of long-term frozen storage on the dynamics of water and ice in wheat bread

Frozen storage of bread has a substantial impact on the dynamics of water and ice in the crumb and crust. In this study, the impact was characterized using wheat bread stored at −18 °C for a long term of ∼4 months. The frozen bread incurred a considerable loss of the crumb water that migrated out and formed ice crystals on the bread surface. Such a moisture decrease underwent more rapidly for the bread stored without intact crust, suggesting the specific role of crust during frozen storage. Moisture also redistributed significantly within the frozen crumb, resulting in an elevated crumb heterogeneity of freezable water. This redistribution of freezable water was accompanied by a progressive recrystallization of the crumb-borne ice crystals, which were measured to grow into bulk sizes using a modified calorimetric procedure for analyzing the crumb samples at their as-frozen states.     

Staling of frozen partly and fully baked breads. Study of the combined effect of amylopectin recrystallization and water content on bread firmness

Partly baked (PB) and fully baked (FB) breads were frozen at −18 °C for 7, 21, 63, 92, 126 and 188 d and were analysed after its thawing (FB) or thawing and final baking (PB). The starch retrogradation, the moisture content and the firmness were measured as properties closely related to the aging of bread. The temperature of glass transition of the maximally freeze-concentrated state, T_g′, was also measured and established in (−18 ± 0.8) °C. This value cannot ensure molecular immobility in both types of bread during its frozen storage at (−18 ± 2) °C. Consequently, the rearrangements of starch component molecules, needed for its recrystallization and for the diffusion of water during frozen storage, could take place and could justify the changes observed in the bread. PB bread showed a significant decrease in firmness with frozen storage, while the firmness of the FB bread did not change significantly, although an increase when compared with the control, not frozen bread, was detected. A regression study led to the conclusion that the combined effect of starch component crystallization and water loss could explain the firming evolution and that both variables exerted an effect of similar intensity on crumb firmness.     

Volume change of bread and bread crumb during cooling,chilling and freezing,and the impact of baking

During baking, bread dough undergoes an expansion followed by a slight contraction at the end of baking. The contraction during baking has been evidenced by some authors. However, there is a limited amount of literature about the contraction of the crumb during the chilling phase and also during the freezing phase in the case of freezing. A study has been carried out to better understand the impact of the baking degree on the contraction of the crumb during chilling after baking and during freezing. The volume of the samples has been evaluated with a laser volumeter. Breads (70 g dough) were baked until reaching 75 °C, 85 °C, 95 °C, 98 °C and then 98 °C for 10 min. Results showed that a longer baking resulted in a lower contraction of the bread. The volume change was between 25% and 2.5% for baking at 75 °C—0 min dwell and 98 °C—10 min dwell, respectively. The contraction was compared to the contraction of degassed bread crumb samples, which was more important. SEM pictures showed that the degree of baking also corresponded to a very different structure of the crumb. For the longer baking, the starch granules were fully gelatinized and no ghosts of starch granules were visible. The magnitude of the contraction was thus associated with the degree of baking and with the degree of starch granule destructuration.     

Rheological properties and baking performance of rye dough as affected by transglutaminase

Since protein aggregation and formation of a continuous protein matrix in rye dough is very limited, an enzyme-induced protein aggregation method to improve the baking properties was investigated. The effects of microbial transglutaminase (TG) on the properties of rye dough were studied by rheological tests, confocal laser scanning microscopy (CSLM), standard-scale baking tests and crumb texture profile analysis. Addition of TG in the range of 0-4000 Ukg⁻¹ rye flour modified the rheological properties of rye flour dough, resulting in a progressive increase of the complex shear modulus (|G∗|) and in a decrease of the loss factor (tan δ) due to protein cross-linking or aggregation. CLSM image analysis illustrated a TG-induced increase of the size of rye protein complexes. Standard baking tests showed positive effects on loaf volume and crumb texture of rye bread with TG applied up to 500 Ukg⁻¹ rye flour. Higher levels of TG (500 U ≤ TG ≤ 4000 U) had detrimental effects on loaf volume. Increasing TG concentration resulted in an increase of crumb springiness and hardness. In conclusion, the results of this work demonstrated that TG can be used to improve the bread making performance of rye dough by creating a continuous protein network.     

Experimental data and modeling of the thermodynamic properties of bread dough at refrigeration and freezing temperatures

Thermodynamic properties of bread dough (fusion enthalpy, apparent specific heat, initial freezing point and unfreezable water) were measured at temperatures from −40 °C to 35 °C using differential scanning calorimetry. The initial freezing point was also calculated based on the water activity of dough. The apparent specific heat varied as a function of temperature: specific heat in the freezing region varied from (1.7–23.1) J g⁻¹ °C⁻¹, and was constant at temperatures above freezing (2.7 J g⁻¹ °C⁻¹). Unfreezable water content varied from (0.174–0.182) g/g of total product. Values of heat capacity as a function of temperature were correlated using thermodynamic models. A modification for low-moisture foodstuffs (such as bread dough) was successfully applied to the experimental data.     

Combining local pressure and temperature measurements during bread baking: insights into crust properties and alveolar structure of crumb

Improvements in both the miniaturisation and heat compensation of pressure transducers made it possible to measure pressures as low as 5 kPa inside bread dough during baking (ΔT = 80 °C). Additional calibration was found to be necessary to decrease it below 0.18 kPa according to the variations in temperature encountered during baking. Two probes with both a thermocouple and a miniature pressure transducer were used to reveal pressure gradients inside bread dough during baking and post-cooling. During baking, increase in pressure (up to 1.1 kPa) was mainly attributed to the mechanical restrictions exerted on the dough by the stiffened surface layers. Pressure build-up due to the stiffening of bubble walls could not be detected. Various effects of the rupture in the bubble walls are reported. Sudden falls in pressure observed up to 70 °C were attributed to the bubble coalescence phenomenon. Evidence of an open porous structure was provided by the balance in pressure through the dough before the end of baking and the almost simultaneous lowering of pressure (−0.15 kPa) throughout the crumb during cooling. The slight lowering of pressure during post-cooling was also evidence of lower permeability of the crust compared to the crumb.     

In situ observation of the freezing process in wheat dough by confocal laser scanning microscopy (CLSM): Formation of ice and changes in the gluten network

Freezing of bread dough is widely applied in food industry. However, freezing impairs the baking performance of dough, which is largely attributed to structural changes as induced by ice formation. The aim of the present investigation was to image ice formation during freezing of dough and to assess the structural changes in the gluten network. A confocal laser scanning microscope (CLSM) equipped with a freezing stage was used to follow ice formation in the reflection and transmission (bright field) mode. Wheat dough with air inclusions served as a model for fermented dough. The gas pores and the ice crystals could be imaged by confocal laser reflection. Ice formation was initiated at the gas pore interface, where large ice crystals were formed during a freezing time of 4 h at −15 °C. The freezing of gluten samples stained with rhodamin was followed in the fluorescence mode. The cryoconcentration of gluten could be observed, but no irreversible changes in the microstructure of gluten were detected upon thawing. It is concluded that the gas pore interfaces in dough are preferential sites for ice nucleation, favouring the growth of ice crystals in these regions and by this a freeze induced redistribution of water in dough.     

Evaluation of water holding capacity and breadmaking properties for frozen dough containing ice structuring proteins from winter wheat

The effects of ice structuring proteins (ISPs) from white wheat and storage conditions on the water holding capacity (WHC) and breadmaking properties of frozen dough were investigated. The WHC of frozen dough was measured by centrifugation and the breadmaking properties were assessed as proofing time and bread specific volume. It was found that the prolonged frozen storage and freeze–thaw cycles decreased the WHC and breadmaking properties of dough. ISPs were highly effective in increasing the WHC of frozen dough and improving the breadmaking properties. There was a strong correlation between WHC and breadmaking properties (proofing time and bread specific volume) of frozen dough.     

Interrelation between mechanical and biological aeration in starch-based gluten-free dough systems

Alternative aeration and gas stabilization strategies are required for the production of starch-based cellular food systems, such as gluten-free bread. In the present study, density and temperature were monitored in mixing experiments without yeast, aiming at maximum mechanical aeration. Additionally, the same trials were performed with subsequent biological aeration, including yeast fermentation and baking. As a result, the gas volume fraction was elevated to 21%, instead of 6% with conventional kneading. Reducing the water content from 120% to 90% (flour/starch weight base) raised dough viscosity and temperature without affecting the state of aeration. The bread volume was strongly influenced by the dough temperature after mixing (R² = 0.98), since it depended on yeast activity. The implemented process is suitable to aerate starch-based dough systems mechanically and enables the production of gluten-free bread with high volume and fine pores.     

Impact of baking conditions and storage temperature on staling of fully and part-baked Sangak bread

Bread staling involves a combination of physico-chemical phenomena that leads to a reduction of quality. This study aims at evaluating the impact of baking conditions (280 °C, 8 min; 310 °C, 5.5 min; 340 °C, 4 min), baking type (of fully baked (FB) and part-baked (PB)) and storage temperature (−18, 4 and 20 °C) on the staling of Sangak bread. Results showed that lower baking temperature with longer baking time produced drier bread with higher firmness. In FB Sangak breads, amylopectin retrogradation, amount of unfreezable water and firmness (measured by compression test) increased during storage at positive temperatures but hardness (determined by Kramer shear test) decreased significantly during first day of storage. The recrystallized amylopectin traps the free water resulting in crumb hardening. Water is also absorbed by the dry crust resulting in changes of rheological properties in the crust and crumb, and finally in staling. Storage at 4 °C resulted in increasing melting enthalpy of amylopectin crystallite in comparison with storage at 20 °C. Also it was found that firmness of PB breads due to rebaking was significantly lower than FB breads. There were no significant changes in staling parameters of FB and PB stored at −18 °C.     

Fate of ACE-inhibitory peptides during the bread-making process: Quantification of peptides in sourdough,bread crumb,steamed bread and soda crackers

This study compared the concentration of angiotensin-converting enzyme (ACE) inhibitory peptides at different stages of the bread-making process, including kneading, proofing, and final products. Steamed bread, baked bread, and soda crackers were produced with 3–20% addition of rye malt sourdoughs to assess products differing in their thermal treatment. Eight tripeptides with known or predicted ACE-inhibitory activity were quantified by LC/MS in multiple reaction monitoring (MRM) mode. In wheat sourdough and rye-malt gluten sourdough, IPP was the predominant tripeptide at 58 and 473 μmol kg⁻¹, respectively, followed by LQP, IQP, and LPP. During the bread-making process, peptide concentrations were modified by enzymatic conversions at the dough stage and by thermal reactions during baking. The concentrations of IPP, LPP and VPP remained stable during dough preparation but decreased during thermal treatment; the concentrations of other peptides were changed at the dough stage but remained relatively stable during baking. The cumulative concentration of 8 ACE-inhibitory peptides in steamed bread and bread crumb exceeded 60 μmol kg⁻¹, while soda crackers contained less than 3 μmol kg⁻¹. The peptide levels in bread thus likely meet in vivo active concentrations.     

Formation and distribution of ice upon freezing of different formulations of wheat bread

The formation and distribution of ice upon the freezing of fresh breadcrumb were investigated using differential scanning calorimetry. Three types of wheat bread containing different amounts of sugar and dietary fiber were measured. Various frozen states were produced through freezing with different cooling rates (0.5, 1, 2, 5, 10, 20 and 30 °C/min) to −30 °C; they were then analyzed and compared by thawing with the same heating rate (10 °C/min) to 20 °C. All DSC heating traces exhibited dual endotherms in the temperature range for the melting of ice: The major transition was attributed to the ice formed in the large crumb pores (gas cells) and the minor event, which preceded the major endotherm, was assigned primarily to the ice formed in the nanometer-sized pores within the gluten-starch matrix. The size of ice crystals in the two classes of pores was estimated using the modified Gibbs–Thompson relation. The distributions of ice in these pores depended on the bread compositions. It is concluded that the complex crumb porosity plays an essential role in shaping the activities of water and ice in the breadcrumb.     

The final established physicochemical properties of steamed bread made from frozen dough: Study of the combined effects of gluten polymerization,water content and starch crystallinity on bread firmness

The gluten polymerization behavior, water content, starch crystallinity and firmness of Chinese steamed bread made from frozen dough were investigated and their correlations were also established in this study. The decreased degree of gluten polymerization in steamed bread was observed by the enhanced SDS-extractable proteins (SDSEPs) upon frozen storage. Less incorporation of glutenin in the glutenin–gliadin crosslinking of steamed bread mainly contributed to the decreased degree of gluten polymerization. The decreased moisture of steamed bread had a significant negative correlation with the sublimated water in frozen dough (r = −0.8850, P < 0.01). Frozen storage also induced an increase in starch crystallinity and bread firmness. A multiple linear regression model with SDS-extractable proteins, water content and melting enthalpy of starch crystals of steamed bread accounted for 86% of the variance in the natural logarithm of firmness and further revealed that starch crystallinity mainly contributed to bread firmness.     

Phytate negatively influences wheat dough and bread characteristics by interfering with cross-linking of glutenin molecules

The influence of added phytate on dough properties and bread baking quality was studied to determine the role of phytate in the impaired functional properties of whole grain wheat flour for baking bread. Phytate addition to refined flour at a 1% level substantially increased mixograph mixing time, generally increased mixograph water absorption, and reduced the SDS-unextractable protein content of dough before and after fermentation as well as the loaf volume of bread. The added phytate also shifted unextractable glutenins toward a lower molecular weight form and increased the iron-chelating activity of dough. It appears that phytate negatively affects gluten development and loaf volume by chelating iron and/or binding glutenins, and consequently interfering with the oxidative cross-linking of glutenin molecules during dough mixing. Phytate could be at least partially responsible for the weak gluten network and decreased loaf volume of whole wheat flour bread as compared to refined flour bread.     

Effect of the thermostable xylanase B (XynB) from Thermotoga maritima on the quality of frozen partially baked bread

The effect of the recombinantly produced xylanase B (XynB) from Thermotoga maritima MSB8 on the quality of frozen partially baked bread (FPBB) was investigated. Addition of XynB to wheat flour dough resulted in a significant increase in dough extensibility (L), swelling (G), and a decrease in dough resistance to deformation (P), configuration. Bread crumb characteristics were studied by differential scanning calorimeter (DSC) and dynamic-mechanical analysis (DMA). The results show that addition of XynB leads to improvements in the bread quality of FPBB and retards bread staling compared to the control. The greatest improvements were obtained in specific volume (+35.2%) and crumb firmness (−40.0%). The control FPBB was significantly firmer in texture and higher in amylopectin recrystallization than the bread with XynB. During frozen storage of FPBB with and without XynB for 8 weeks, the crumb firmness increased gradually and the specific volume slightly decreased with the frozen storage time. The ΔH values of freezable water (FW) endothermic transitions increased with frozen storage time for all samples. However, addition of XynB lowered the ΔH values indicating a decrease in FW. Therefore, XynB is useful in improving the quality of FPBB. DMA was also used to monitor the shrinking behavior of the samples. Addition of XynB increased the contraction during chilling but significantly diminished the total shrinking and frozen-state shrinking of the bread crumb during the freezing process.     

Frozen bread dough: Effects of freezing storage and dough improvers

This review focuses on the effects of freezing storage on the microstructure and baking performance of frozen doughs, and provides an overview of the activities of dough improvers, including emulsifiers, hydrocolloids and other improvers used in frozen dough applications. The overall quality of bread baked from frozen dough deteriorates as the storage of the dough at sub-zero temperatures increases due to several factors which are discussed. Lipid-related emulsifiers such as diacetyl tartaric acid esters of mono and diglycerides and sucrose esters employed as anti-staling agents, dough modifiers, shortening sparing agents, and as improvers for the production of high-protein bread have also been employed in frozen doughs. Hydrocolloids are gaining importance in the baking industry as dough improvers due to their ability to induce structural changes in the main components of wheat flour systems during breadmaking steps and bread storage Their effects in frozen doughs is discussed. Other dough improvers, such as ascorbic acid, honey and green tea extract, are also reviewed in the context of frozen doughs.