Gluten hydrolysis and depolymerization during sourdough fermentation

Hydrolysis and depolymerization of gluten proteins during sourdough fermentation were determined. Neutral and acidified doughs in which microbial growth and metabolism were inhibited were used as controls to take into account the proteolytic activity of cereal enzymes. Doughs were characterized with respect to cell counts, pH, and amino nitrogen concentrations as well as the quantity and size distribution of SDS-soluble proteins. Furthermore, sequential extractions of proteins and analysis by HPLC and SDS-PAGE were carried out. Sourdough fermentation resulted in a solubilization and depolymerization of the gluten macropolymer. This depolymerization of gluten proteins was also observed in acid aseptic doughs, but not in neutral aseptic doughs. Hydrolysis of glutenins and occurrence of hydrolysis products upon sourdough fermentation were observed by electrophoretic analysis. Comparison of sourdoughs with acid control doughs demonstrated that glutenin hydrolysis and gluten depolymerization in sourdough were mainly caused by pH-dependent activation of cereal enzymes.     

脱酰胺与双酶协同作用提高小麦面筋蛋白酶解效率

为了探讨了不同脱酰胺处理和双酶协同作用方式对小麦面筋蛋白酶解效率及其产物抗氧化活性的影响,该文研究了小麦面筋蛋白在各种预处理方式和酶解条件下的蛋白回收率、水解度、抗氧化性能及肽分子量分布情况。结果显示,单独热处理(90℃,30 min)小麦面筋蛋白对其酶解效率无显著影响,而采用添加0.5 mol/L柠檬酸溶液进行热处理(质量分数为5%,90℃,30 min)可显著(P0.05)提高其蛋白回收率。此外,酶制剂添加顺序及双酶共同水解作用时间对酶解效率均具有较大影响:加入谷氨酰胺酶预先水解对小麦面筋蛋白的深度水解有促进作用;一定时间内的双酶协同作用有利于酶解的进行,但较长时间的双酶作用反而会抑制酶解效率。采用谷氨酰胺酶(质量分数为0.2%)对经柠檬酸加热处理的小麦面筋蛋白作用12 h后再加入胰酶(质量分数为0.6%)共同作用7 h可使蛋白回收率达70.74%,水解度达到9.88%;另外,酶解产物的自由基清除能力ABTS+(2,2’-Azinobis-(3-ethylbenzthiazoline-6-sulphonate)+)值与氧化自由基吸收能力(ORAC,oxygen radical absorbance capacity)值分别达到478.95 mmol/g和213.85μmol/g,提示该酶解产物是一种潜在优秀食品抗氧化剂。研究结果可为拓宽小麦面筋蛋白的应用领域,以及高效制备抗氧化活性肽提供方法和理论指导。     

基于管式模型的小麦面筋系统流变行为的解析

利用动态流变仪对8种小麦面筋系统的进行小形变试验,研究在振动模式下的频率扫描时,面筋系统流变学特性,从分子动力学的角度进行解析,并引入Pierre Gilles de Gennes等人的管式模型理论基础性地对面筋流变行为进行阐述。试验表明：面筋蛋白转变温度在52.6～58.5℃间;0.1～40℃间,频率扫描下,时间-温度叠加法算出20℃时表观活化能在4602.57～6511.16 J/mol间,不同样品间有较大的差异;Arrhenius模型和WLF模型在拟合面筋系统叠合曲线的效果上一致。通过SPSS分析相关性表明：在面筋蛋白转变温度下,固定频率的温度扫描的弹性模量和在不同温度下的频率扫描的弹性模量成显著的正相关;面筋蛋白的转变温度与表观活化能呈较显著的正相关。该试验结果为小麦面筋相关制品的开发提供了参考。     

Important aroma compounds in freshly ground wholemeal and white wheat flour-identification and quantitative changes during sourdough fermentation

An investigation on the odor-active compounds of wholemeal (WWF) and white wheat flour (WF 550) by aroma extract dilution analysis (AEDA) and by quantitative studies using stable isotope dilution assays (SIDA) revealed a significant number of odor-active compounds, such as (E)-2-nonenal, (E,Z)- and (E,E)-2,4-decadienal, (E)-4,5-epoxy-(E)-2-decenal, 3-hydroxy-4,5-dimethyl-2(5H)-furanone, and vanillin, with high odor activities in both wheat flours. The amounts and, consequently, the aroma potencies of vanillin, (E,E)-2,4-decadienal, and 3-(methylthio)propanal were much higher in the WWF than in the WF 550 samples. Fermentation of suspensions of both flours with lactic acid bacteria did not generate new odorants; however, many compounds, such as acetic acid or 3-methylbutanal, were increased, whereas aldehydes (formed from the degradation of unsaturated fatty acids) were decreased. Comparing the odorant concentrations present before and after fermentation gave evidence that the main influence of the microorganisms on sourdough aroma is to either enhance or decrease specific volatiles already present in the flour. A comparison with literature data indicated that most of these odorants are also important for the bread crumb aroma present after baking of the dough.     

Prolonged fermentation of whole wheat sourdough reduces phytate level and increases soluble magnesium

This work was designed to compare the effects of different leavens (yeast, sourdough, and a mixture of both) on phytic acid (PA) degradation and to assess the repercussions of PA breakdown on phosphorus and magnesium solubility during bread-making. Sourdough fermentation was more efficient than yeast fermentation in reducing the phytate content in whole wheat bread (-62 and -38%, respectively). Furthermore, lactic acid bacteria present in sourdough enhanced acidification, leading to increased magnesium and phosphorus solubility. To intensify phytate breakdown, bran was incubated with microorganisms (yeast or sourdough) before bread-making. Using this new method, the percentage of phytate breakdown was near 90%, whereas 40% of phytate remained in traditional French bread. In conclusion, a prolonged fermentation with sourdough still leads to improved Mg and P solubility by decreasing phytate content and through acidification.     

Kinetics of transglutaminase-induced cross-linking of wheat proteins in dough

The effects of TGase in dough after 15, 30, 45, and 60 min of resting time after mixing were studied with a Kieffer test. The resistance to stretching of control dough did not change greatly during the 60 min time period after mixing. In dough, TGase decreased extensibility and increased resistance to stretching and this change was already observed after the first 15 min (first measurement). The higher the enzyme dosage was, the higher the magnitude of the rheological change was. All of the doughs that contained TGase 3.8 or 5.7 nkat/g flour had a higher resistance to stretching and lower extensibility than control dough 15 min after mixing. Resistance to stretching clearly increased at a dosage of 5.7 nkat/g flour during the 15-60 min period after mixing. Extensibility increased in the control dough and in the doughs with a low enzyme dosage almost at the same rate. The evolution of air bubbles during proofing was determined with bright field microscopy and image analysis. In the presence of 5.7 nkat/g TGase, the fermented dough contained more of the smallest and less large air bubbles in comparison to the control dough. The effect of TGase and water content on the specific volume of the conventional and organic wheat bread was studied. Water did not have a significant effect on the specific volume of bread. TGase increased the specific volume of breads baked from organic flour only, when additional water (+10% of farinogram absorption) and a small enzyme dosage were used. Microstructural characterization showed that bread baked without TGase from conventional flour had a stronger protein network than that baked from organic flour. TGase improved the formation of protein network in breads baked from either normal or organic flour but at higher dosage caused uneven distribution.     

Evidence for formation of heterooligosaccharides by Lactobacillus sanfranciscensis during growth in wheat sourdough

Lactobacillus sanfranciscensis is a key organism of the lactic microflora in traditional and industrial sourdough fermentations. In this paper we provide evidence for the formation of heterooligosaccharides (HeOS) by L. sanfranciscensis during growth in sourdough. To identify the HeOS based on HPAEC-PAD analysis, HeOS standards were synthesized by enzymatic reactions with L. sanfranciscensis levansucrase in a chemically defined system in the presence of raffinose, maltotriose, maltose, xylose, or arabinose as acceptor carbohydrates. The oligosaccharides known to originate from the corresponding acceptor reactions, 1(F)-beta-fructosylraffinose, 1(F)-beta-fructofuranosylmaltotriose, erlose (1(F)-beta-fructofuranosylmaltose), xylsucrose, 1(F)-beta-fructosylxylsucrose, and arabsucrose, were identified by HPAEC-PAD. Evidence for the formation of further tri-, tetra-, and pentasaccharides was provided. Wheat doughs with sucrose were fermented with L. sanfranciscesis TMW 1.392 or the isogenic, levansucrase-negative strain TMW 1.392Deltalev, and the analysis of dough extracts or invertase-treated dough extracts provided evidence for the formation of arabsucrose and erlose in sourdough in addition to 1-kestose and nystose.     

Mechanisms of heat-mediated aggregation of wheat gluten protein upon pasta processing

During pasta processing, structural changes of protein occur, due to changes in water content, mechanical energy input, and high temperature treatments. The present paper investigates the impact of successive and intense thermal treatments (high temperature drying, cooking, and overcooking) on aggregation of gluten protein in pasta. Protein aggregation was evaluated by the measurement of sensitivity of disulfide bonds toward reduction with dithioerythritol (DTE), at different reactions times. In addition to the loss in protein extractability in sodium dodecyl sulfate buffer, heat treatments induced a drastic change in disulfide bonds sensitivity toward DTE reduction and in size-exclusion high-performance liquid chromatography profiles of fully reduced protein. The protein solubility loss was assumed to derive from the increasing connectivity of protein upon heat treatments. The increasing degree of protein upon aggregation would be due to the formation of additional interchain disulfide bonds.     

加工方式对非发酵面团小麦醇溶蛋白致敏性的影响

该研究探究了热加工(水煮、烘烤、微波、高温高压)、冷处理(液氮、速冻机速冻)和非热加工(超高静压、辐照、脉冲强光、臭氧和超声波)处理对非发酵面团小麦醇溶蛋白致敏性的影响。利用SDS-PAGE和双抗体夹心ELISA法研究各种加工处理后非发酵面团小麦醇溶蛋白的分子量和致敏性变化。结果表明:200和300 MPa的超高静压处理后,非发酵面团小麦醇溶蛋白的致敏性显著增加(P<0.05),均增至125%左右;水煮、微波、高温高压、烘烤、液氮、速冻机处理、超高静压250MPa、辐照(7、13kGy)、臭氧熏蒸、脉冲强光和超声波处理均能显著降低非发酵面团小麦醇溶蛋白的致敏性(P<0.05),其中以水煮、高温高压、液氮、臭氧和脉冲强光(PL-1)处理的脱敏效果最显著(P<0.01),均降至60%左右。由此可知,加工方式能够显著影响非发酵面团小麦醇溶蛋白的致敏性,可作为食品中过敏原安全控制的有效手段,为脱敏食品的生产提供有效参考。进一步研究需要结合其他体内试验的评估,特别是小麦易敏人群的临床试验。     

The importance of lactic acid bacteria for phytate degradation during cereal dough fermentation

Lactic acid fermentation of cereal flours resulted in a 100 (rye), 95-100 (wheat), and 39-47% (oat) reduction in phytate content within 24 h. The extent of phytate degradation was shown to be independent from the lactic acid bacteria strain used for fermentation. However, phytate degradation during cereal dough fermentation was positively correlated with endogenous plant phytase activity (rye, 6750 mU g(-1); wheat, 2930 mU g(-1); and oat, 23 mU g(-1)), and heat inactivation of the endogenous cereal phytases prior to lactic acid fermentation resulted in a complete loss of phytate degradation. Phytate degradation was restored after addition of a purified phytase to the liquid dough. Incubation of the cereal flours in buffered solutions resulted in a pH-dependent phytate degradation. The optimum of phytate degradation was shown to be around pH 5.5. Studies on phytase production of 50 lactic acid bacteria strains, previously isolated from sourdoughs, did not result in a significant production of intra- as well as extracellular phytase activity. Therefore, lactic acid bacteria do not participate directly in phytate degradation but provide favorable conditions for the endogenous cereal phytase activity by lowering the pH value.     

小麦粉面团形成过程水分状态及其比例变化

为揭示小麦粉面团形成过程水分状态和比例、面团结构的变化,以及这种变化与粉质仪和拉伸仪表征的质量特性之间的关系;认识面团形成过程表征筋力强弱的物质基础和变化机理。选用中筋（宁春4号）和强筋（师栾02-1）小麦品种为试验材料,利用低场核磁共振技术测定粉质仪和面过程、拉伸仪醒发拉伸过程不同时间点面团水分状态和比例的变化;利用红外显微成像技术分析面团形成过程不同取样点蛋白质和淀粉的分布及结构变化。结果表明,面粉原料中主要为弱结合水。面粉在粉质仪加水搅拌形成面团后,水分状态和比例发生显著变化,面团中的水可以分为强结合水（T21）、弱结合水（T22）和自由水（T23）。面团搅拌形成过程中,中筋小麦品种宁春4号面团中的强结合水比例显著降低;师栾02-1的强结合水的弛豫时间在和面终点消失,弱结合水的弛豫时间显著延长,而自由水的比例显著增加（P＜0.05）。强筋小麦粉强结合水的保持时间较长。拉伸过程加盐和不加盐对同一取样点、同一种水分状态之间的水分弛豫时间和比例无显著影响;宁春4号自由水的弛豫时间在加盐和不加盐处理时都显著缩短（P＜0.05）。湿面筋含量高、筋力较强面团的蛋白质网络结构致密。粉质仪和面过程强结合水和弱结合水弛豫时间和比例的变化,与面筋含量和强度有关。该结论可为面制品加工过程和面工艺选择与优化等方面提供一定的理论参考。     

Simultaneous determination of hydrolysis and mutarotation rates during the enzymatic hydrolysis of lactose

An experiment is described in which a custom-made glucose electrode is used to directly monitor the enzymatic hydrolysis of lactose to glucose. The transient profile of beta- d-glucose can be used to simultaneously determine the rate constants for mutarotation and for enzymatic hydrolysis by applying a dynamic nonlinear regression routine. Due to differences in the mutarotation rate constants between lactose and glucose, the beta- d-glucose concentration "overshoots" equilibrium under certain conditions, which can be modeled mathematically. This overshoot can be observed reliably and used to quantify the differences in mutarotational equilibria between glucose and lactose. These observations may be important for the analysis of dairy products and commercial lactase preparations and illustrate an unusual kinetic phenomenon caused by intramolecular forces. This approach may also be important for the accurate determination of a variety of oligosaccharides such as glycogen, which tend to be composed primarily of one stereoisomer.     

Modifications of wheat flour proteins during in vitro digestion of bread dough, crumb, and crust: an electrophoretic and immunological study

The proteins of wheat flour have several biological activities that can affect human health and physiology when wheat-based foods are consumed. The modifications of bread crumb and crust proteins during an in vitro peptic/pancreatic digestion process were studied by electrophoresis and immunoblotting with polyclonal antibodies specific for single proteins or groups of homologous proteins of the wheat flour, and the results were compared to those obtained for an unheated dough sample. The results show that baking affects the extent of proteolysis and the immunological and physicochemical features of the digestion products in relation to the level of the heat treatment. Therefore, the results concerning the digestion of the unheated wheat flour or dough are not representative of what happens when baked products enter the human digestive tract.     

Water-extractable and water-unextractable arabinoxylans affect gluten agglomeration behavior during wheat flour gluten-starch separation

Water-extractable arabinoxylan (WE-AX) of variable molecular weight (MW) and water-unextractable arabinoxylan (WU-AX) were added to wheat flour to study their effect on gluten agglomeration in a dough and batter gluten-starch separation process with recovery of gluten from the batter with a set of vibrating sieves (400, 250, and 125 microm). Low MW WE-AX had almost no impact on the distribution of the gluten on the different sieves. High MW WE-AX decreased yields of the largest (400 microm sieve) gluten aggregates, more than their medium MW counterparts, indicating the importance of AX MW for their effect on gluten interactions. Correlations between the total level of gluten protein recovered on the three sieves and the batter extract viscosity as well as between the proportion of gluten protein recovered on the 400 microm sieve to that on the three sieves and the batter extract viscosity pointed to the importance of viscosity as an indicator for gluten agglomeration, as did the fact that another viscosity increasing plant polysaccharide (guar gum) also negatively influenced gluten agglomeration. However, the obtained data cannot rule out that AX and guar gum also exert steric effects on gluten agglomeration. WU-AX, present as discrete cell wall fragments, had a negative impact on the level of large gluten aggregates. Taken together, the results show that both native WE-AX and WU-AX detrimentally impact gluten agglomeration.     

Physicochemical behaviors of sugars, lipids, and gluten in short dough and biscuit

The structure of short dough and biscuit has been characterized at a macroscopic level (dimensions, bulk structure) and a microscopic level (starch damage, protein aggregates, microstructure) by physical and biochemical methods. The baking process of short dough induces a large decrease of the product bulk density from 1.26 to 0. 42 (+/-0.01) g.cm(-)(3) for final biscuit, leading to a cellular solid with a thin colored surface and a porous inner structure. Proteins appear aggregated in biscuit when compared to short dough, whereas starch granules remain almost intact in biscuits. The components which are involved in the cohesiveness of short dough and biscuit final structure have been identified. They suggest that short dough is a suspension of solid particles in a liquid phase being an emulsion of lipids in a concentrated sugar solution. The role of sugars in biscuit structure suggest that biscuit structure is a composite matrix of protein aggregates, lipids and sugars, embedding starch granules.     

Moderate decrease of pH by sourdough fermentation is sufficient to reduce phytate content of whole wheat flour through endogenous phytase activity

Whole wheat bread is an important source of minerals but also contains considerable amounts of phytic acid, which is known to impair their absorption. An in vitro trial was performed to assess the effect of a moderate drop of the dough pH (around 5.5) by way of sourdough fermentation or by exogenous organic acid addition on phytate hydrolysis. It was shown that a slight acidification of the dough (pH 5.5) with either sourdough or lactic acid addition allowed a significant phytate breakdown (70% of the initial flour content compared to 40% without any leavening agent or acidification). This result highlights the predominance of wheat phytase activity over sourdough microflora phytase activity during moderate sourdough fermentation and shows that a slight drop of the pH (pH value around 5.5) is sufficient to reduce significantly the phytate content of a wholemeal flour. Mg "bioaccessibility"of whole wheat dough was improved by direct solubilization of the cation and by phytate hydrolysis.     

Properties of chemically and physically treated wheat gluten films

Chemical (vapors of formaldehyde), physical (temperature, UV and gamma radiation), and aging treatments were applied to wheat gluten films. Changes in film mechanical properties, water vapor permeability, solubility, and color coordinates were investigated. An aging of 360 h led to a 75 and 314% increase in tensile strength and Young's modulus, respectively, and a 36% decrease in elongation. Severe thermal (above 110 degrees C, 15 min) and formaldehyde treatments highly improved the mechanical resistance of the films. Under these conditions, up to 376 and 654% increase in tensile strength and Young's modulus and up to 66% decrease in elongation have been observed. Water solubility was only slightly modified, whereas water vapor permeability was not affected. Color coordinates of films heated above 95 degrees C changed to a great extent. An almost total insolubilization of proteins in sodium dodecyl sulfate occurred for heat- and formaldehyde-treated films, due to the modification of protein network leading to changes in properties of the films.