Antifreeze Activity of γ‐Polyglutamic Acid and Its Impact on Freezing Resistance of Yeast and Frozen Sweet Dough

This study investigated the antifreeze activity (AF) of γ‐polyglutamic acid (γ‐PGA), freezing resistance of yeast cells and sweet dough, and the mechanism influenced by γ‐PGA. Properties studied included AF of γ‐PGA, water‐holding capacity of flour, survival ratio and oxidation resistance capability of yeast cells, ice melting enthalpy (ΔH), and fermentation and breadmaking properties of sweet dough. The AF of γ‐PGA was 8.03 g of unfrozen water/g of sample, indicating good AF. γ‐PGA was tested on yeast cells and sweet dough stored frozen for 0, 1, 2, 4, and 8 weeks at four levels (0, 0.5, 1, and 3%). Survival ratio of yeast cells with γ‐PGA was significantly higher than the corresponding control. A possible mechanism might be related to the modulation of oxidation resistance capability of yeast cells by γ‐PGA. A decrease in glutathione release from frozen yeast cells and an increase in water‐holding capacity of wheat dough were observed with the addition of γ‐PGA. In the presence of γ‐PGA, ΔH, ice melting temperature, and proofing time of frozen sweet dough decreased significantly, and fermentation parameters improved, compared with the corresponding control sample. Specific volume of bread made from frozen sweet dough with 0.5, 1, and 3% γ‐PGA increased by 6.3, 8.9, and 3.3%, respectively, after 8 weeks of frozen storage. γ‐PGA enhanced the freezing resistance of yeast cells and sweet dough effectively, and the effect on specific volume of bread was not linear, with 1% showing better results.     

Metabolite Analysis Allows Insight into the Differences in Functionality of 25 Saccharomyces cerevisiae Strains in Bread Dough Fermentation

During dough fermentation, yeast (Saccharomyces cerevisiae) changes the physical properties of the dough matrix. In this study, we investigate if different yeast strains have an impact on dough rheology and on the gas holding capacity of fermenting dough. Furthermore, we analyze whether observed differences are linked to the metabolite profiles of the yeast strains. More specifically, the impact of 25 yeast strains on dough spread, dough fermentation properties, and dough metabolite profile was analyzed. Our results demonstrate large differences in the fermentation ability and metabolite profile of the 25 strains. Analysis of metabolites in fermented dough confirmed that acetic acid and succinic acid are likely responsible for the lowering of dough pH during fermentation and that the onset of CO₂ release from dough is related to dough pH rather than to the volume of CO₂ within the dough. Our results further suggest that the spread test is an inadequate tool to quantify rheological differences observed for strains with different fermentation profiles.     

酵母对冷冻面团发酵特性及馒头品质的影响

为了找出使冷冻面团和馒头综合品质较稳定的酵母产品,采用动态流变仪和F3发酵仪对低糖型国光高活性干酵母(1#)、英联马利苹果即发高活性干酵母(2#)、品一高活性干酵母(3#)、高糖型马利即发高活性干酵母(4#)、高糖型丹宝利即发高活性干酵母(5#)和耐高糖安琪高活性干酵母(6#)冷冻面团冻藏35 d过程中的流变学特性和发酵特性进行研究,并对由此面团制作馒头的质构、色泽、比容和感官品质进行分析。结果表明:不同市售酵母冷冻面团在不同冻藏时间下的流变学特性和发酵特性不同。不同酵母冷冻面团制作馒头后硬度、弹性、回复性、咀嚼性、亮度、红度、黄度、比容和感官品质分别差异显著,不同冻藏时间下同种酵母冷冻面团馒头的质构、色泽、比容和感官品质也分别差异显著。6种酵母在冻藏35 d内,1#和2#酵母冷冻面团的发酵活力始终较大,6#酵母冷冻面团的发酵特性参数始终最稳定;1#、5#和6#酵母冷冻面团的弹性模量与黏性模量较大,其中面团流变学特性最稳定的是5#;1#和6#酵母冷冻面团制作馒头的感官品质较好的同时,比容较高,色泽品质较好,质构品质也较好。因此整个冻藏期间,使冷冻面团和馒头综合品质较好较稳定的是1#,其次是6#。研究结果为冷冻面团馒头工业化生产中酵母的选择提供参考。     

Dynamic and Elongation Rheology of Yeasted Bread Doughs

The rheology of yeasted bread doughs is a little‐studied field despite yeast's importance in developing bread structure. A method of thermally inactivating the yeast within mixed bread doughs was developed to overcome the difficulty of yeast fermenting during rheological measurements. Sample stabilization by preshearing of dough samples at a stress amplitude of 1 Pa at 1 Hz for 10 sec improved the reliability of small amplitude oscillatory shear measurements, and resting 20 min within the rheometer was sufficient to produce reliable and consistent observations. Small amplitude oscillatory shear measurements were unable to detect any differences between yeasted and nonyeasted doughs nor any changes in linear viscoelastic properties due to fermentation. However, large strain uniaxial elongation measurements of yeasted doughs revealed a significant progressive decrease in elongational viscosities with fermentation. Size‐exclusion HPLC analysis of yeasted doughs showed an increase in unextractable polymeric dough proteins, which were interpreted as evidence of cross‐linking and therefore a potential improvement in dough properties. The apparent contradictions between uniaxial elongation and SE‐HPLC studies of fermenting yeasted doughs can be attributed to gas bubbles within the dough interrupting the increasingly cross‐linked protein network, resulting in the rheological weakness observed for fermenting yeasted doughs.     

Heterologous expression of type I antifreeze peptide GS-5 in baker's yeast increases freeze tolerance and provides enhanced gas production in frozen dough

The demand for frozen-dough products has increased notably in the baking industry. Nowadays, no appropriate industrial baker's yeast with optimal gassing capacity in frozen dough is, however, available, and it is unlikely that classical breeding programs could provide significant improvements of this trait. Antifreeze proteins, found in diverse organisms, display the ability to inhibit the growth of ice, allowing them to survive at temperatures below 0 degrees C. In this study a recombinant antifreeze peptide GS-5 was expressed from the polar fish grubby sculpin (Myoxocephalus aenaeus) in laboratory and industrial baker's yeast strains of Saccharomyces cerevisiae. Production of the recombinant protein increased freezing tolerance in both strains tested. Furthermore, expression of the GS-5 encoding gene enhanced notably the gassing rate and total gas production in frozen and frozen sweet doughs. These effects are unlikely to be due to reduced osmotic damage during freezing/thawing, because recombinant cells showed growth behavior similar to that of the parent under hypermosmotic stress conditions.     

复合改良剂对后发酵馒头冷冻面团冻藏品质的影响

冷冻面团加工技术作为一种面食生产新工艺,是目前较先进的面制品保鲜技术。冷冻处理会对酵母活力、面团品质、面团面筋结构等产生不良影响。为了提高冷冻面团品质,明确不同食品改良剂对后发酵冷冻面团馒头品质的影响,该研究优化了复配增稠剂(卡拉胶、瓜尔豆胶、海藻酸钠)、复配乳化剂(双乙酰酒石酸单双甘油酯、羧甲基纤维素)、复配酶制剂(谷氨酰胺转氨酶、葡萄糖氧化酶)和海藻糖对后发酵冷冻面团馒头品质的作用,通过单因素和响应面试验设计,考察了不同复合改良剂对冷冻面团馒头比容、质构和感官品质的影响。研究表明:海藻糖、酶制剂对冷冻面团馒头比容的影响极其显著(p0.01)。在添加量为0.4%乳化剂、0.8%增稠剂、45 mg/kg酶制剂、2%海藻糖的复配工艺下,冷冻面团馒头的比容达到最佳水3.15 mL/g,其硬度和弹性也达到较优值,分别为926.832 g、0.912 8。同时对未添加和添加复合改良剂冻藏0～2.5个月的冷冻面团进行扫描电镜观察,发现未添加复配改良剂的冷冻面团中大多数小淀粉颗粒裸露在外,颗粒分明,内部组织结构不均匀,而添加复合改良剂的冷冻面团淀粉颗粒镶嵌在面筋网络之间,蛋白网络结构清晰完整,冷冻面团内部组织结构均匀紧密,面团的抗冻性较强。     

Effects of Yeast Freezing in Frozen Dough

The effects of freezing and frozen storage of bread dough and compressed yeast on bread quality were studied. Besides, the effects of compressed yeast freezing on cell viability, gas production and release of substances by the yeast cells were examined. Freezing and frozen storage of dough made with fresh yeast had more negative effects on baking quality than the addition of frozen yeast to dough. When the compressed yeast is frozen and stored at ‐18°C, the CO₂ production decreased, while the amount of dead cells, the total protein, and the total reducing substances leached from the yeast increased as the length of yeast frozen storage increased. SDS‐PAGE showed that the substances leached from frozen yeast caused an increase in the solubility of some gluten proteins. On the other hand, size‐exclusion chromatography (SEC) pointed out that the relative amount of two protein fractions of low molecular weight leached from frozen yeast increased for longer yeast frozen storage periods. The yeast leachates had an adverse effect on loaf volume.     

Impact of Sodium Alginate and Xanthan Gum on the Quality of Steamed Bread Made from Frozen Dough

The impact of freezing on dough rheology, fermentation performance, and final steamed bread quality was investigated in this study. Also, the incorporation of sodium alginate and xanthan gum into the frozen dough formulation, in comparison with 0.1% salt, was studied to test their suitability as frozen dough improvers. Incorporating these hydrocolloids into steamed bread revealed their totally different characteristics from those in baked bread. Freezing of dough led to diminished specific volumes of proofed dough and steamed bread, and it also caused higher crumb firmness for steamed bread. The incorporation of sodium alginate and xanthan gum did not improve the quality of the steamed bread but led to further reduction in specific volume and increase in crumb firmness at the higher levels of 0.3, 0.5, and 1.0% and 0.07 and 0.1%, respectively. Xanthan gum and sodium alginate showed dough strengthening effects by increasing resistance to uniaxial deformation, bubble burst stress, and declining dough weakening coefficients at these levels, but decreased dough extensibility and bubble burst strain were revealed at these concentrations tested.     

The Combination of Rhizopus chinensis Lipase and Transglutaminase Affects the Rheology and Glutenin Macropolymer Properties of Frozen Dough

The combination of Rhizopus chinensis lipase (RCL) and transglutaminase (TG) was previously reported to improve the quality of frozen dough bread. In this study, the effects of RCL, TG, and their combination on the modification of glutenin macropolymer (GMP) and rheological properties of dough during frozen storage were investigated. Frozen storage changed both GMP and rheology properties of dough. TG treatment significantly decreased the ratio of high‐molecular‐weight glutenin subunits to low‐molecular‐weight glutenin subunits and GMP content in fresh dough, and GMP particle size increased. The effect of RCL on GMP properties was not significant, but its combination with TG dramatically increased the proportion of the larger particles and weighted average volume (D_4.3) in GMP. The treatment with the enzyme combination could have inhibited the depolymerization of GMP, which slowed down the decrease rate of some parameters such as GMP content, proportion of larger particles, D_4.3, and release of free amino and thiol groups during frozen storage. The modification of GMP properties by enzyme treatment weakened the effect of the freezing process on rheological properties of dough, especially TG treatment and its combination with RCL. Correlation between GMP particle size and dough properties (dough tensile force and elastic modulus) after freezing and enzyme treatment were confirmed.     

Properties of Arabinoxylans in Frozen Dough Enriched with Wheat Fiber

The global market for frozen bread dough is rising; however, its quality could deteriorate during extended storage. Our previous study indicated that undesirable changes caused by freezing could be reduced by adding arabinoxylan‐rich fiber sources. The present study investigated the changes in arabinoxylan properties of yeasted dough during frozen storage. Dough samples made from refined, whole, and fiber‐enriched (15% either wheat aleurone or bran) flours were stored at –18°C for nine weeks, and structural properties of arabinoxylan were probed during storage. Water‐extractable arabinoxylan (WEAX) content in dough samples increased by about 19–33% during the first three weeks of storage. Prolonged storage of dough (weeks 6 and 9), however, correlated with a decline in WEAX content. Average molecular weight and intrinsic viscosity of WEAX decreased during storage for all frozen dough samples. Arabinose‐to‐xylose ratios also decreased by 11 and 6% for control and composite dough samples, respectively. There was a significant positive correlation (r = 0.89, P < 0.0001) between WEAX content of dough and bread quality throughout the storage period. The results demonstrated that changes in dough quality during frozen storage were related to changes in the content and structure of WEAX that took place during frozen storage.     

Validation of antifreeze properties of glutathione based on its thermodynamic characteristics and protection of baker's yeast during cryopreservation

The antifreeze ability of glutathione was evaluated on the basis of its thermodynamic characteristics and protection of baker's yeast during cryopreservation at -30 degrees C. The thermodynamic characteristics and protection of baker's yeast of glutathione were similar to those of known antifreeze proteins, such as carrot antifreeze protein and holly antifreeze protein. These properties included lowering the freezing point at about 0.20 degrees C non-colligatively, decreasing freezable water content, controlling the movement of free water for its strong hydrophilicity, and improving baker's yeast survival during the simulated processing of frozen dough. Therefore, glutathione was viewed to be an antifreeze protein like substance on the basis of its unique thermodynamic characteristics and protection of baker's yeast. The method combining thermodynamic characteristic analysis and protection evaluation is a new and simple way to screen new antifreeze proteins.     

Effect of freezing and frozen storage of doughs on bread quality

The effects of freezing and storage in frozen conditions on bread quality, crumb properties, and aggregative behavior of glutenins were analyzed. The effect of different additives on bread quality was also studied. The results obtained showed that freezing and storage at -18 degrees C decreased the bread quality. Samples stored in frozen conditions supplemented with diacetyl-tartaric acid ester of monoglycerides, gluten, and guar gum produced breads of greater volume and more open crumb structure than those prepared with the base formulation (without additives). All additives analyzed increased the proof time. Crumb firmness increased with dough frozen storage and bread aging time at 4 degrees C. A decrease in the amount of glutenin subunits of high molecular mass was observed by electrophoresis analysis of the SDS-soluble proteins aggregates extracted from the frozen dough. This result suggested that the protein matrix of bread underwent depolymerization during storage in frozen conditions.     

可移动双孢菇栽培料发酵车设计及效果试验

在中国多数地区,双孢菇以家庭小规模栽培生产为主,由于双孢菇培养料发酵达不到要求,双孢菇存在产量低、效益差等问题,是双孢菇未得到大面积推广栽培的因素之一。可移动式发酵车是针对中小规模双孢菇栽培者使用的研发产品,主要由通风底座、发酵室、压力风机、控制系统、通风管道、控制阀、换热器等部分构成。根据原料性状,完成一次发酵需要15~20 d,发酵过程中无需翻堆倒料;发酵料上、中、下三层温差可以控制在0.1~3.9℃,均质程度高;发酵料色泽、手感、秸秆纤维、气味、浸出液等物理性状均符合堆肥腐熟的判定标准,且栽培双孢菇平均产量能达到18 kg/m~2。     

Fermentation Stability and Pore Size Distribution of Frozen Prefermented Lean Wheat Doughs

Fermentation stability of frozen prefermented doughs was studied with a maturograph, an instrument that allows monitoring of dough rise, gas production, and gas retention during fermentation. Maturograph curves excellently predicted the baking quality, measured as form ratio, after frozen storage. The greatest decrease in dough level occurred after seven days of storage, after which the level remained constant. With some flours, decreased amount of water improved both the fermentation stability and form ratio of breads baked after seven days of frozen storage of dough. However, no improvement was observed in loaf volume. Preliminary experiments with longer final fermentation time (after thawing) showed that the reduced water content also resulted in higher loaf volumes than did optimal water content. Microscopic studies showed that with most doughs, porosity decreased with reduced water content. However, these changes depended on flour type. In one dough, reduction of water by 2 percentage units decreased the area of pores per total area of section from 56.6 to 46.4%, whereas in another dough the same water reduction had no effect on the pore area.     

Effect of Hydrophilic Gums on the Quality of Frozen Dough: Electron Microscopy,Protein Solubility,and Electrophoresis Studies

Hydrophilic gums have been shown to improve the shelf‐life stability of frozen doughs during long periods of frozen storage. The objective of this research was to determine the effect of gums on starch and protein characteristics of frozen doughs using electron microscopy and electrophoresis studies. Frozen doughs, supplemented with three levels of gum arabic, carboxy methyl cellulose (CMC), kappa (κ) carrageenan, and locust bean gum, were studied after day 1 and after 4, 8, 12, and 16 weeks of frozen storage. Changes in the ultra structure of the frozen doughs were investigated, as well as the solubilities and composition of dough proteins by SDS‐PAGE. Scanning electron micrographs of doughs evaluated on day 0 (unfrozen) showed starch granules securely embedded in the gluten matrix. However, after 8 and 16 weeks of frozen storage, the frozen control dough without the gum additives clearly showed damage to the gluten network, and the starch granules appeared to be separated from the gluten. Doughs with locust bean gum and gum arabic showed better retention of the gluten network compared with the frozen control evaluated after different periods of storage. The SDS‐soluble protein content increased while residue protein content decreased as the frozen storage time increased. After each frozen storage period, the control dough without the gum additive had the highest amount of SDS‐soluble proteins and the lowest amount of residue proteins when compared with the doughs treated with gums. κ‐Carrageenan and locust bean gum had the lowest amount of SDS‐soluble proteins compared with doughs with CMC and gum arabic. The frozen control had the lowest amount of residue proteins at any particular time of frozen storage. κ‐Carrageenan treated doughs had the highest amount of residue proteins, followed by doughs with locust bean gum. Doughs with gum arabic and CMC had the lowest amount of residue proteins but still higher than the control doughs.     

Correlations Between Empirical and Fundamental Rheology Measurements and Baking Performance of Frozen Bread Dough

Empirical and fundamental rheology measurements were made on fresh and frozen dough to investigate the effects of freezing, frozen storage, and additives. These results were compared with results of a standard baking test. Four formulations were tested: a control dough, and doughs with additions of 100 ppm of ascorbic acid (AA), 0.5% sodium stearoyl lactylate (SSL), and 0.5% diacetyl tartaric acid esters of monoglycerides (DATEM). Rheological and baking tests were performed on fresh doughs and on doughs after two, five, and eight weeks of frozen storage. Resistance to extension was higher for doughs with additives in fresh and frozen doughs. There was a decrease in resistance to extension due to freezing. Complex modulus in fresh doughs was highest for doughs with SSL. There was a decrease complex modulus after freezing and thawing. In frozen doughs at 10 Hz, doughs with additives had higher complex modulus values and lower phase angle values when compared to the control. The additives used all had a positive effect on proof time, loaf volume, and crumb firmness, and all formulations deteriorated in quality during frozen storage. Resistance to extension and complex modulus were positively correlated with loaf volume (r = 0.86 and r = 0.64, P < 0.01). Phase angle was negatively correlated with loaf volume (r = -0.74, P < 0.01).     

Improvement of texture properties and flavor of frozen dough by carrot (Daucus carota) antifreeze protein supplementation

The effects of concentrated carrot protein (CCP) containing 15.4% (w/w) carrot (Daucus carota) antifreeze protein on texture properties of frozen dough and volatile compounds of crumb were studied. CCP supplementation lowered the freezable water content of the dough, resulting in some beneficial effects including holding loaf volume steadily and making the dough softer and steadier during frozen storage. Furthermore, SPME-GC-MS analysis showed CCP supplementation did not give any negative influences on volatile compounds of crumb and gave a pleasant aroma felt like Michelia alba DC from trans-caryophyllene simultaneously. Combining our previous results that CCP supplementation improves the fermentation capacity of the frozen dough, CCP could be used as a beneficial additive for frozen dough processing.     

液氮冷媒介质对速冻饺子冻裂率的影响

研究饺子新型冻结技术,探索液氮速冻工艺对调理食品品质的作用规律。该文以饺子为研究对象,和面水面质量比、皮馅质量比、面粉品质、馅料配比、产品规格、冻结时间、冻结方式、冻结介质等诸多因素对饺子冻裂率都有重要影响。该试验以猪肉、香菇、大豆组织蛋白为饺子馅主要原料,研究了和面时的水面质量比、面团的静置时间、皮馅比、饺子在液氮池中的沉浸时间对饺子冻裂率的影响。结果表明:以上四因素对饺子的冻裂率都有不同程度的影响。和面水面质量比为56∶100、面团静置时间在10 min、皮馅质量比在1∶1.3、液氮池沉浸时间为40 s,制作好的饺子在立即速冻后的冻裂率为37%,感官评定得分为77.6。该研究为液氮沉浸速冻工艺在调理食品的应用提供了一定的理论参考。     

Effect of Hydrophilic Gums on Frozen Dough. I. Dough Quality

Disadvantages of frozen doughs are their variable performance and loss of stability over long‐term frozen storage. Changes in rheological properties of frozen doughs have been reported to be due to the physical damage of the gluten network caused by ice crystallization and recrystallization. The objective of this study was to determine the effect of hydrophilic gums on ice crystallization and recrystallization for improvement of the shelf‐life stability of frozen dough. The present research involved use of the Hard Red Spring wheat cultivar Grandin and hydrophilic gums such as carboxymethyl cellulose (CMC), gum arabic, kappa carrageenan (κ‐carrageenan), and locust bean gum at three different levels each on doughs stored frozen for up to 16 weeks. The dough characteristics were analyzed after day 0, day 1, and after 4, 8, 12, and 16 weeks of frozen storage using data from differential scanning calorimetry (DSC), water activity, extensigraph, and proof time. The ΔH value of freezable water endothermic transitions obtained using DSC increased with storage time for all treatments. However, addition of different levels of the four gums lowered the ΔH value, indicating a decrease in freezable water. Doughs with locust bean gum gave a higher peak force, measured using the Kieffer dough extensibility rig of the texture analyzer, and lower proof time, indicating better retention of baking quality. Maximum resistance to extension increased upon addition of 1 and 3%; CMC; 1 and 3%; κ‐carrageenan; and 1, 2, and 3% locust bean gum as compared with the control. The various periods of storage or gum treatments did not affect the water activity of the thawed frozen doughs. Doughs with locust bean gum gave significantly lower proof time compared with the other treatments and the control. CMC gave the second lowest values, followed by gum arabic treatment. Addition of κ‐carrageenan increased the proof time compared with the control. In summary, locust bean gum, gum arabic, and CMC improved the dough characteristics to varying degrees. κ‐Carrageenan was the only gum that showed a detrimental effect on frozen dough.     

Changes in Phytate Content in Whole Meal Wheat Dough and Bread Fermented with Phytase‐Active Yeasts

The degradation of inositol hexakisphosphate (IP6) was evaluated in whole meal wheat dough fermented with baker's yeast without phytase activity, different strains of Saccharomyces cerevisiae (L1.12 or L6.06), or Pichia kudriavzevii with extracellular phytase activity to see if the degradation of IP6 in whole meal dough and the corresponding bread could be increased by fermentation with phytase‐active yeasts. The IP6 degradation was measured after the dough was mixed for 19 min, after the completion of fermentation, and in bread after baking. Around 60–70% of the initial value of IP6 in the flour (10.02 mg/g) was reduced in the dough already after mixing, and additionally 10–20% was reduced after fermentation. The highest degradation of IP6 was seen in dough fermented with the phytase‐active yeast strains S. cerevisiae L1.12 and P. kudriavzevii L3.04. Activity of wheat phytase in whole meal wheat dough seems to be the primary source of phytate degradation, and the degradation is considerably higher in this study with a mixing time of 19 min compared with earlier studies. The additional degradation of IP6 by phytase‐active yeasts was not related to their extracellular phytase activities, suggesting that phytases from the yeasts are inhibited differently. Therefore, the highest degradation of IP6 and expected highest mineral bioavailability in whole meal wheat bread can be achieved by use of a phytase‐active yeast strain with less inhibition. The strain S. cerevisiae L1.12 is suitable for this because it was the most effective yeast strain in reducing the amount of IP6 in dough during a short fermentation time.