New Saccharomyces cerevisiae baker's yeast displaying enhanced resistance to freezing

Three procedures were used to obtain new Saccharomyces cerevisiae baker's yeasts with increased storage stability at -20, 4, 22, and 30 degrees C. The first used mitochondria from highly ethanol-tolerant wine yeast, which were transferred to baker's strains. Viability of the heteroplasmons was improved shortly after freezing. However, after prolonged storage, viability dramatically decreased and was accompanied by an increase in the frequency of respiratory-deficient (petite) mutant formation. This indicated that mitochondria were not stable and were incompatible with the nucleus. The strains tested regained their original resistance to freezing after recovering their own mitochondria. The second procedure used hybrid formation after protoplast fusion and isolation on selective media of fusants from baker's yeast meiotic products resistant to parafluorphenylalanine and cycloheximide, respectively. No hybrids were obtained when using the parentals, probably due to the high ploidy of the baker's strains. Hybrids obtained from nonisogenic strains manifested in all cases a resistance to freezing intermediate between those of their parental strains. Hybrids from crosses between meiotic products of the same strain were always more sensitive than their parentals. The third method was used to develop baker's yeast mutants resistant to 2-deoxy-d-glucose (DOG) and deregulated for maltose and sucrose metabolism. Mutant DOG21 displayed a slight increase in trehalose content and viability both in frozen doughs and during storage at 4 and 22 degrees C. This mutant also displayed a capacity to ferment, under laboratory conditions, both lean and sweet fresh and frozen doughs. For industrial uses, fermented lean and sweet bakery products, both from fresh and frozen doughs obtained with mutant DOG21, were of better quality with regard to volume, texture, and organoleptic properties than those produced by the wild type.     

Mathematical model of sugar uptake in fermenting yeasted dough

Fermentation prior to freezing significantly reduces the shelf life of frozen dough, measured as a decline in proofing power. Changes during fermentation caused by yeast metabolism have previously been described empirically on a dough weight basis and have not been mathematically modeled. In this work, yeast metabolites were quantified in fermenting dough and their concentrations were estimated in the aqueous environment around yeast cells. The osmotic pressure in the aqueous phase increases by 23% during 3 h of fermentation, which depresses the freezing point by 1 degrees C. The rise in osmotic pressure and the accumulation of ethanol may affect phase equilibria in the dough, baking properties, and the shelf life of frozen dough. Predictive modeling equations fitted sugar concentration data accurately. It was found that the preference of baker's yeast for glucose over fructose was stronger in fermenting dough than in liquid fermentations. The usefulness of the model in industrial bakery formulation work was demonstrated.     

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.     

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.     

Improved organoleptic and nutritive properties of bakery products supplemented with amino acid overproducing Saccharomyces cerevisiae yeasts

Spontaneous yeast mutants isolated in continuous culture as resistant to toxic amino acid analogues, able to increase up to 40 times their free amino acid pool of Thr, up to 160 times their pool of Met, or up to 20 times their pool of Lys, were characterized with regard to properties of industrial interest. Growth rate, mu (h(-1)), and biomass yield, Y (g/L), of the amino acid overproducing mutants (AA(S)) were in many cases similar to those of the wild type, whereas their free amino acid content was substantially increased in laboratory and industrial media (molasses). Doughs fermented with 3% baker's yeast and 0.5% AA(S) mutants produced bakery products that displayed texture similar to those fermented with 3.5% baker's yeast, but the former had a considerable improvement of their taste and aroma. On the other hand, bread content of the essential amino acids Lys, Met, and Thr provided by yeast was also increased.     

Respiratory burst after freezing and thawing of soil: Experiments with soil bacteria

The respiratory burst after freezing and thawing of soil has its maximum immediately after thawing. The reduction in bacterial plate counts after a freeze-thaw cycle was moderate, and so was also the subsequent increase. Experiments with isolated bacteria from the same soil showed that there was a similar burst when bacteria were frozen and thawed in the stationary phase. Between different strains, there was a positive correlation between the killing effect of the freeze-thaw treatment and the respiratory increase per surviving bacterium. The increased O₂ uptake during the first 11 h after thawing corresponded to a C consumption 5–15% of that in the killed bacteria. A brief respiratory increase was observed when the supernatant from a frozen-thawed culture was added to bacteria in the stationary state. The burst was more persistent when frozen-thawed bacteria were centrifuged and then resuspended in supernatants from stationary cultures of unfrozen and frozen bacteria, or even in a mineral salts solution. The optical difference spectrum between the supernatants from frozen and unfrozen cultures demonstrated leakage of cellular material after freezing and thawing. Monod kinetics for growth shows that if the substrate concentration is substantially lower than K_s, and the biomass concentration relatively high, the respiratory rate is at maximum at the time when the substrate is added. The respiratory burst following freezing and thawing can thus be explained on the basis of simple growth kinetics.     

Generation of thiols by biotransformation of cysteine-aldehyde conjugates with baker's yeast

Baker's yeast was shown to catalyze the transformation of cysteine-furfural conjugate into 2-furfurylthiol. The biotransformation's yield and kinetics were influenced by the reaction parameters such as pH, incubation mode (aerobic and anaerobic), and substrate concentration. 2-Furfurylthiol was obtained in an optimal 37% yield when cysteine-furfural conjugate at a 20 mM concentration was anaerobically incubated with whole cell baker's yeast at pH 8.0 and 30 degrees C. Similarly to 2-furfurylthiol, 5-methyl-2-furfurylthiol (11%), benzylthiol (8%), 2-thiophenemethanethiol (22%), 3-methyl-2-thiophenemethanethiol (3%), and 2-pyrrolemethanethiol (6%) were obtained from the corresponding cysteine-aldehyde conjugates by incubation with baker's yeast. This work indicates the versatile bioconversion capacity of baker's yeast for the generation of thiols from cysteine-aldehyde conjugates. Thanks to its food-grade character, baker's yeast provides a biochemical tool to produce thiols, which can be used as flavorings in foods and beverages.     

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.     

Development of a simplified method for the determination of folates in baker's yeast by HPLC with ultraviolet and fluorescence detection

A simplified HPLC method for rapid determination of folates in yeast with ultraviolet and fluorescence detection without sample purification has been developed. By use of the column Aquasil C(18), specially designed for polar analytes, and gradient elution, it was possible to separate and determine five folate derivatives: tetrahydrofolate, 5-methyltetrahydrofolate, and 5-formyltetrahydrofolate with fluorescence detection, and 10-formylfolic acid and folic acid with ultraviolet detection. The sample preparation required only a small amount of dry yeast (25-50 mg) and included an extraction of folates by heat treatment and deconjugation of folate polyglutamates to monoglutamates with the use of rat serum conjugase. Validation involved investigation of matrix effects, determination of recovery by standard addition method, repeatability, and stability tests. The dominating folate forms in commercial dry baker's yeast were found to be tetrahydrafolate and 5-methyltetrahydrofolate with a total folate content of 2890 microg/100 g (63.4 nmol/g). The simplicity of the method makes it suitable for folate screening studies of different yeast strains.     

土体冻结过程中基质势与水分迁移及冻胀的关系

土体冻结过程中不同位置液态水的能量差引起了水分迁移与重分布,进而引发冻胀,关于势能差驱动下的冻土水分迁移问题一直由于技术手段的匮乏而没有完全解决。利用新近推出的可用于冻土水热研究的p F meter基质势传感器与5TM水分传感器,实时监测研究饱和青藏红黏土单向冻结过程中基质势-液态含水率-温度-含冰量-水分迁移量-冻胀变形之间在时间、空间上的耦合变化关系。结果表明:土体温度场变化引起内部液态水相变,打破了原有的能量平衡,试验结束后12~14 cm土样高处含水率最高达到55%,靠近冻融交界面处(10 cm)的未冻区含水率减小至25.8%,水分整体向冷端发生迁移;土体冻胀的快慢及冻胀量大小与水分迁移速率及数量具有线性关系;试验后土体内总含水率的分布与分凝冰透镜体的分布一致,已冻区液态含水率的分布与温度梯度近似成线性关系,未冻区液态含水率的分布与水分的迁移量有关,与温度梯度无关。此外,温度场对水分场的变化具有诱导作用但二者并不同步,当冻结速率减小到一定程度时水分才开始迁移,第10小时后温度场趋于稳定而水分迁移并未停止。研究成果揭示了土体单向冻结过程中液态水、基质势、温度等物理参数的动态变化过程及内在联系,为冻胀机制的研究以及冻胀模型的建立提供了试验基础。     

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。     

Inositol hexaphosphate hydrolysis by Baker's yeast. Capacity, kinetics, and degradation products

Phytases hydrolyze myo-inositol 1,2,3,4,5,6-hexaphosphate (IP(6)), yielding lower inositol phosphates and inorganic orthophosphate. Two commercial strains of baker's yeast (Saccharomyces cerevisiae), Y(1) and Y(2), were able to express phytase activity. This was determined by the capacity to grow in a synthetic medium with IP(6) as the sole phosphorus source. IP(6) hydrolysis was rapid for both strains, and after 24 h, all IP(6) was degraded. Control cultures contained inorganic orthophosphate (P(i)) and no IP(6). Growth rate in IP(6) medium was for both strains essentially identical to growth in P(i) medium, indicating a well-adapted metabolism for utilization of phosphorus from IP(6). There was some difference in growth yield (milligrams of biomass per milligram of glucose) between the two strains: 0.95 (Y(1)) and 1.35 (Y(2)) in IP(6) medium and 1.03 and 1. 35, respectively, in P(i) medium. The phytases were of the 3-phytase type, forming mainly DL-Ins(1,2,4,5,6)P(5), DL-Ins(1,2,5,6)P(4), and DL-Ins(1,2,6)P(3).     

Expression of a bacterial ice nucleation gene in a yeast Saccharomyces cerevisiae and its possible application in food freezing processes

A 3.6 kb ice nucleation gene (ina) isolated from Erwinia herbicola was placed under control of the galactose-inducible promoter (GAL1) and introduced into Saccharomyces cerevisiae. Yeast transformants showed increased ice nucleation activity over untransformed controls. The freezing temperature of a small volume of water droplets containing yeast cells was increased from approximately -13 degrees C in the untransformed controls to -6 degrees C in ina-expressing (Ina(+)) transformants. Lower temperature growth of Ina(+) yeast at temperatures below 25 degrees C was required for the expression of ice nucleation activity. Shift of temperature to 5-20 degrees C could induce the ice nucleation activity of Ina(+) yeast when grown at 25 degrees C, and maximum ice nucleation activity was achieved after induction at 5 degrees C for approximately 12 h. The effects of Ina(+) yeast on freezing and texturization of several food materials was also demonstrated.     

Stereospecificity of myo-inositol hexakisphosphate dephosphorylation by a phytate-degrading enzyme of baker's yeast

During food processing such as baking, phytate is dephosphorylated to produce degradation products, such as myo-inositol pentakis-, tetrakis-, tris-, bis-, and monophosphates. Certain myo-inositol phosphates have been proposed to have positive effects on human health. The position of the phosphate groups on the myo-inositol ring is thereby of great significance for their physiological functions. Using a combination of high-performance ion chromatography analysis and kinetic studies the stereospecificity of myo-inositol hexakisphosphate dephosphorylation by a phytate-degrading enzyme from baker's yeast (Saccharomyces cerevisiae) was established. The data demonstrate that the phytate-degrading enzyme from baker's yeast dephosphorylates myo-inositol hexakisphosphate in a stereospecific way by sequential removal of phosphate groups via D-Ins(1,2,4,5,6)P(5), D-Ins(1,2,5,6)P(4), D-Ins(1,2,6)P(3), D-Ins(1,2)P(2), to finally Ins(2)P (notation 3/4/5/6/1). Knowledge of the absolute stereochemical specificity of the baker's yeast phytase allows use of the enzyme to produce defined myo-inositol phosphates for kinetic and physiological studies.     

磁场辅助冻结对冷冻熟制面条品质的影响

在食品的冻结方式中,磁场是一种新兴的物理方法,其特点是在食品原料中具有很强的穿透性。为改善冷冻熟制面条的品质,探究了磁场辅助冻结对冷冻熟制面条品质的影响。选取不同的磁场强度：3、6、9、12、15、18 Gs,辅助冻结面条。结果表明：磁场强度为12 Gs时,与未加磁场的空白组相比,冷冻熟制面条的冻结时间显著（P<0.05）缩短了4 min、白度显著（P<0.05）增大2.37%、硬度显著（P<0.05）提升7.40%、咀嚼性显著（P<0.05）增大12.41%、剪切力显著（P<0.05）增大19.20%、蒸煮损失显著（P<0.05）降低17.88%、微观结构中冰晶更细小、可冻结水含量显著（P<0.05）降低9.44%、水分分布状态中结合水含量显著（P<0.05）提升10.95%、淀粉分子的峰强度比值降低,老化程度显著（P<0.05）降低0.22%,结果表明,磁场辅助冻结可以改善冷冻熟制面条的品质,12 Gs的磁场强度冻结效果较好。     

Determination of cellular carbohydrates in peanut fungal pathogens and baker's yeast by capillary electrophoresis and electrochromatography.

In this work, the quantitation of cellular carbohydrates, namely chitin and glucan, in peanut fungal pathogens and baker's yeast was carried out by capillary electrophoresis (CE) and capillary electrochromatography (CEC). The chitin and glucan of the fungi were hydrolyzed by the enzymes chitinase and glucanase, respectively, to their corresponding sugar monomers N-acetylglucosamine (GlcNAc) and glucose (Glc). These two monosaccharides were then tagged with 6-aminoquinoline (6-AQ) to allow their separation and detection in CE and CEC. The 6-AQ derivatives of GlcNAc and Glc formed the basis for the determination by CE and CEC of chitin and glucan in peanut fungi and baker's yeast. Several parameters affecting the separation of the 6-AQ derivatives of GlcNAc and Glc, including the separation voltage and the composition of the running electrolyte, were investigated. Under the optimized separation conditions, the contents of cellular carbohydrates including N-acetylglucosamine, chitin, glucose, and glucan in some fungi, such as Sclerotinia minor, Sclerotium rolfsii, and baker's yeast, were successfully determined. The method described here allowed the assessment of genetic differences in Sclerotium rolfsii isolates from various locations.     

大同市季节性冻土的分布与变化特征

利用大同市1961-2008年8个站点地面气象观测记录中的冻土资料,对大同市地面冻结日期、解冻日期、冻结日数及最大冻土深度的时空分布进行分析。结果表明:40多年来土壤开始冻结期呈推迟的趋势,土壤完全解冻期呈提前的趋势,地面冻结日数相应呈减少趋势,累年最大冻土深度自西北向东南逐渐减小,全市历年平均最大冻土深度变化总体上呈减少趋势,但2000年以后较20世纪60-80年代有增加的趋势。     

Variation in fatty acid composition of Artemia salina nauplii enriched with microalgae and baker's yeast for use in larviculture

The high content of the essential fatty acids in some microalgae and baker's yeast has made them excellent diets for boosting the fatty acid content of livefood Artemia. The influences of baker's yeast (Saccharomyces cerevisiae) and three microalgae, viz., Chlorella salina, Chaetoceros calcitrans, and Nannochloropsis salina, were tested as diet components in marine livefeed brine shrimp Artemia salina nauplii to improve the polyunsaturated fatty acid (PUFA) composition. Artemia nauplii submerged in these diets for four different enrichment intervals (3, 6, 8, and 24 h) were found to incorporate essential fatty acids, and the percentage composition of different fatty acids was measured in the enriched Artemia nauplii and enrichment diets. N. salina produced higher levels of arachidonic acid (AA, 20:4n6, 9.50%), eicosapentaenoic acid (EPA, 20:5n3, 25.80%), and docosahexaenoic acid (DHA, 22:6n3, 4.18%) as compared to other diets. The total PUFA content of the enriched Artemia by N. salina increased by 56.50% with enrichment periods up to 8 h, followed by a significant reduction in the final 24 h. N. salina yielded Artemia nauplii with considerable EPA (8.05%), AA (14.15%), and DHA (1.85%) after 8 h of enrichment, which are significantly higher levels than in nauplii fed with the other three diets (p = 0.05). The DHA/EPA values in Artemia enriched for 6 h by N. salina and C. calcitrans were found to be, respectively, 88.46 and 25% higher than freshly hatched Artemia. Artemia enriched by C. salina and baker's yeast exhibited a reduction in PUFA content even at 6 h of enrichment. Significant relative decreases in DHA, EPA, and total PUFA in Artemia enriched with all of the diets were apparent, with a corresponding increase in the total saturated fatty acid content (26.95 +/- 9.75%) in the final stages (24 h) of enrichment (p = 0.05).     

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.