FLOCCULATION MECHANISMS OF TOP AND BOTTOM FERMENTING BREWING YEAST

The flocculation behaviour of a large set of top and bottom fermenting brewing yeasts was investigated. Bottom and top fermenting strains flocculated according to different mechanisms. Bottom strains flocculated in the stationary growth phase in the presence of sufficiently high Ca²⁺ and sufficiently low sugar concentrations; these strains possessed a lectin-mediated flocculation mechanism. Top strains flocculated in the stationary growth phase without addition of Ca²⁺, only in the presence of sufficiently high concentrations of ethanol. Some of the top strains were inhibited with mannose, but not with sucrose or galactose, while others were not inhibited by any of these sugars. The different sensitivity of flocculation of top and bottom strains with respect to ethanol may be related to the hydrophobicity of the cell surface. Flocculation models for bottom and top fermenting yeasts were proposed. It is suggested that, besides the sugar inhibition pattern, the sensitivity of flocculation with respect to ethanol should be included as an additional parameter for classification of brewing yeasts .     

Adsorption of inorganic and organic ligands onto hydrous aluminum oxide: evaluation of surface charge and the impacts on particle and NOM removal during water treatment

The variable removal from solution of sulfate, orthophosphate, fluoride, five simple organic acids, and natural organic matterfromtwo sources by adsorption on aluminum hydroxide was examined to assess their potential influence on coagulation during drinking water treatment. Measurements of electrophoretic mobility were conducted with adsorption studies to provide means of evaluating the impact of the adsorption of these anions on the removal of particulate material during coagulation at water treatment facilities. The three inorganic ions exhibited widely different trends in terms of removal from solution and effect on the surface charge of the aluminum hydroxide. Phosphate was nearly completely removed from solution across a wide pH range and was observed to lower surface charge and shift the isoelectric point. Sulfate was removed to a lesser extent than phosphate, lowered the surface charge on the precipitate, but did not shiftthe isoelectric point. Fluoride was well-removed through adsorption but exhibited no influence on the charge of the hydrous aluminum oxide. The organic acids similarly displayed varying abilities to be removed through adsorption and different influences on surface charge. The results indicate the importance of the number and location of functional groups and their acid/ base properties. The ability to strongly influence surface charge illustrates the impact that adsorption of these anions can have on particle stability.     

Potential of yeast isolated from apple juice to adhere to stainless steel surfaces in the apple juice processing industry

The aim of this study was to assess the potential of four yeast strains to initiate a biofilm on stainless steel. The yeasts were isolated from apple juice and were identified as Kluyveromyces marxianus, Candida krusei, Zygosaccharomyces sp. and Rhodotorula rubra. The physiochemical properties of cell surface were determined under different conditions. The adhesion capacity of the yeast strains to stainless steel was assayed in presence of apple juice. Cell surfaces were always negatively charged except for Zygosaccharomyces sp., whose isoelectric point was around 3.0. Neither electrophoretic mobility nor flocculation coefficient correlated with the capacity of attachment to stainless steel. The hydrophobicity expressed by the yeast surfaces at pH 3.0, correlated positively with the rate of adhesion (number of cells/min) of each strain. These results indicated that cell surface hydrophobicity governs the initial attachment of the studied contaminant yeast strains to stainless steel surfaces common to the apple juice processing plant.     

Modeling the acid-base properties of bacterial surfaces: A combined spectroscopic and potentiometric study of the gram-positive bacterium Bacillus subtilis

In this study, macroscopic and spectroscopic data were combined to develop a surface complexation model that describes the acid-base properties of Bacillus subtilis. The bacteria were freeze-dried and then resuspended in 0.1 M NaCl ionic medium. Macroscopic measurements included potentiometric acid-base titrations and electrophoretic mobility measurements. In addition, ATR-FTIR spectra of wet pastes from suspensions of Bacillus subtilis at different pH values were collected. The least-squares program MAGPIE was used to generate a surface complexation model that takes into account the presence of three acid-base sites on the surface: tripple bond COOH, tripple bond NH+, and tripple bond PO-, which were identified previously by XPS measurements. Both potentiometric titration data and ATR-FTIR spectra were used quantitatively, and electrostatic effects at the charged bacterial surface were accounted for using the constant capacitance model. The model was calculated using two different approaches: in the first one XPS data were used to constrain the ratio of the total concentrations of all three surface sites. The capacitance of the double layer, the total buffer capacity, and the deprotonation constants of the tripple bond NH+, tripple bond POH, and tripple bond COOH species were determined in the fit. A second approach is presented in which the ratio determined by XPS of the total concentrations of tripple bond NH+ to tripple bond PO- sites is relaxed. The total concentration of tripple bond PO- sites was determined in the fit, while the deprotonation constant for tripple bond POH was manually varied until the minimization led to a model which predicted an isoelectric point that resulted in consistency with electrophoretic mobility data. The model explains well the buffering capacity of Bacillus subtilis suspensions in a wide pH range (between pH=3 and pH=9) which is of considerable environmental interest. In particular, a similar quantitative use of the IR data opens up possibilities to model other bacterial surfaces at the laboratory scale and help estimate the buffering capacity of carboxylate-containing compounds in natural samples.     

猪血蛋白肽功能性质的研究

试验测定了猪血蛋白肽的化学成分和等电点,研究了pH、蛋白肽浓度对猪血蛋白肽溶解性、浊度、乳化性、乳化稳定性、起泡性和起泡稳定性的影响。结果表明:本试验所用猪血蛋白肽蛋白含量为85.03%;等电点为pH5,在等电点时,其溶解性、乳化性、乳化稳定性和起泡性最小,而浊度、起泡稳定性在这点最大;温度在50℃时蛋白肽开始变性,高于50℃时蛋白质的溶解性降低,浊度增加。     

Kinetics of calcium phosphate nucleation and growth on calcite: implications for predicting the fate of dissolved phosphate species in alkaline soils

Unraveling the kinetics of calcium orthophosphate (Ca-P) precipitation and dissolution is important for our understanding of the transformation and mobility of dissolved phosphate species in soils. Here we use an in situ atomic force microscopy (AFM) coupled with a fluid reaction cell to study the interaction of phosphate-bearing solutions with calcite surfaces. We observe that the mineral surface-induced formation of Ca-P phases is initiated with the aggregation of clusters leading to the nucleation and subsequent growth of Ca-P phases on calcite, at various pH values and ionic strengths relevant to soil solution conditions. A significant decrease in the dissolved phosphate concentration occurs due to the promoted nucleation of Ca-P phases on calcite surfaces at elevated phosphate concentrations and more significantly at high salt concentrations. Also, kinetic data analyses show that low concentrations of citrate caused an increase in the nucleation rate of Ca-P phases. However, at higher concentrations of citrate, nucleation acceleration was reversed with much longer induction times to form Ca-P nuclei. These results demonstrate that the nucleation-modifying properties of small organic molecules may be scaled up to analyze Ca-P dissolution-precipitation processes that are mediated by a more complex soil environment. This in situ observation, albeit preliminary, may contribute to an improved understanding of the fate of dissolved phosphate species in diverse soil systems.     

环境条件对猪血浆蛋白功能性质的影响

本实验测定猪血浆蛋白的化学成分和等电点,并研究温度、pH 值、蛋白浓度、离子强度对猪血浆蛋白溶解性、起泡性、泡沫稳定性、乳化性和乳化稳定性质的影响。结果表明：猪血浆蛋白粉中蛋白含量为62.5%;猪血浆蛋白的等电点为pH5,在等电点时,其溶解性、起泡性和乳化稳定性都最小,而起泡稳定性在此点最大;温度在20～70℃范围内,溶解性随温度升高而增大,温度为70℃时开始变性,高于70℃时蛋白质的溶解性降低。一定浓度的食盐的加入对猪血浆蛋白的功能性质起正效应。     

鸡腿菇子实体蛋白提取工艺优化及其特性研究

以鸡腿菇子实体为原料,研究其蛋白的提取工艺及特性。以提取液pH值、提取温度、料液比、提取时间为提取鸡腿菇子实体蛋白的影响因素,采用正交试验优化碱溶酸沉法提取蛋白的工艺条件,结果表明:鸡腿菇子实体蛋白的最佳提取工艺参数为提取液pH11.5,提取温度50℃,料液比1:15,提取时间50min,此时蛋白提取率为24.3%。在pH3.5条件下沉淀蛋白效果最佳。对制备的鸡腿菇子实体蛋白特性的分析表明:其分子质量主要集中在50～90ku,20～35ku和13～17ku等区间内;必需氨基酸(EAA)占氨基酸总量(TAA)的40.57%,必需氨基酸与非必需氨基酸(NEAA)的比值为0.68,为理想蛋白质,且其所含人体必需氨基酸种类齐全,比例均衡,比值系数分(SRC)为80.69;在碱性条件下,鸡腿菇子实体蛋白具有良好的溶解性、乳化性和起泡能力。     

VARIATION OF CELL WALL CONTENT IN FLOCCULENT AND NON-FLOCCULENT YEAST STRAINS

Estimation of the wall content of a number of flocculent and non-flocculent yeast strains taken in both the growth and stationary phases reveals that flocculent yeasts almost invariably increase their wall content to a greater extent as the cells pass to the stationary phase than do non-flocculent strains. This variation of increase is observed whether the yeasts are grown in hopped wort or in a synthetic malt-yeast-peptone-glucose medium. There are considerable variations in wall content from strain to strain in both types of yeast. Cells of flocculent strains are more difficult to disrupt than those of powdery strains. The nitrogen, phosphate and carbohydrate contents of the walls of all strains varied between the growth and stationary phases but no significant trends were noted in any of the components which could be related to flocculent or non-flocculent behaviour.     

Functional properties of acetylated and succinylated sunflower protein isolate

Sunflower protein isolate was acylated with acetic and succinic anhydride. Succinylation changed the electrophoretic mobility, decreased the lysine and chlorogenic acid content and bulk density, increased the aqueous solubility, shifted the isoelectric point and increased the absorption, emulsification and foaming properties. Acetylation improved the functional properties of the protein to a much lesser extent. Implication of these changes is discussed with reference to food applications.     

大豆分离蛋白乳化性的研究

采用蛋白质乳化容量电导法,对不同浓度、ＰＨ和酶水解条件下大豆分离蛋白乳化容量和乳化稳定性进行测定,结果表明：大豆蛋白的乳化性在低密度时随浓度上升而增加,浓度达到６％以后趋于稳定;等电点时（ＰＨ４．５）,乳化性最差,偏离等电点后尤其在偏碱性条件下,乳化性明显增加,酶水解后,乳化性变化产大,水解度１７％时,乳化性最佳。     

Foam properties of algae soluble protein isolate: Effect of pH and ionic strength

In this study the foam properties of algae soluble protein isolate (ASPI), a mixture of mainly proteins and polysaccharides, were investigated as function of isolate concentration (0.1–1.0 mg/mL) and pH (3.0–7.0) at 10 mM and 200 mM NaCl. In addition, adsorption kinetics and dilatational elasticity at the air–water interface were studied. Whey protein isolate (WPI) and egg white albumin (EWA) were used as reference proteins. The consistent dilatational behaviour of ASPI at all pH values and ionic strengths tested indicated a similar interfacial composition at all these conditions. Adsorption kinetics, in contrast, were influenced by varying environmental conditions. At increased ionic strength and close to the theoretical isoelectric point calculated based purely on the amino acid composition of ASPI (pH 7) adsorption increased. Since similar adsorption behaviour was also observed for WPI and EWA, the interfacial properties of ASPI are most likely dominated by its protein fraction. This is further confirmed by the fact that ζ-potential measurements suggested an overall isoelectric point of ASPI below pH 3, while adsorption kinetics varied between pH 5 and pH 7 (the theoretical protein-based isoelectric point of ASPI). The overall foam stability of ASPI stabilized foams was superior to those of WPI and EWA in the pH range 5–7. In conclusion, the molecular and interfacial properties of ASPI, a mixture of proteins and polysaccharides, seem to favour the production of very stable foams in this pH range by the selective adsorption of its protein fraction to the air–water interface.     

Adsorption and aggregation properties of norovirus GI and GII virus-like particles demonstrate differing responses to solution chemistry

The transport properties (adsorption and aggregation behavior) of virus-like particles (VLPs) of two strains of norovirus ("Norwalk" GI.1 and "Houston" GII.4) were studied in a variety of solution chemistries. GI.1 and GII.4 VLPs were found to be stable against aggregation at pH 4.0-8.0. At pH 9.0, GI.1 VLPs rapidly disintegrated. The attachment efficiencies (α) of GI.1 and GII.4 VLPs to silica increased with increasing ionic strength in NaCl solutions at pH 8.0. The attachment efficiency of GI.1 VLPs decreased as pH was increased above the isoelectric point (pH 5.0), whereas at and below the isoelectric point, the attachment efficiency was erratic. Ca(2+) and Mg(2+) dramatically increased the attachment efficiencies of GI.1 and GII.4 VLPs, which may be due to specific interactions with the VLP capsids. Bicarbonate decreased attachment efficiencies for both GI.1 and GII.4 VLPs, whereas phosphate decreased the attachment efficiency of GI.1, while increasing GII.4 attachment efficiency. The observed differences in GI.1 and GII.4 VLP attachment efficiencies in response to solution chemistry may be attributed to differential responses of the unique arrangement of exposed amino acid residues on the capsid surface of each VLP strain.     

热诱导大豆蛋白纤维聚集体的分离及性质研究

酸性热处理条件下,随着大豆分离蛋白(SPI)自发形成纤维聚集体,仍有部分未自组装聚集的多肽分子存在其中,二者比例对蛋白功能性质产生较大影响。本文以超滤手段对热诱导大豆蛋白纤维聚集体和多肽进行大量分离,确定分离方法并研究两者的理化性质。结果表明:100 ku超滤膜反复分离两次即能获得较好的分离效果;SPI经热处理后,其等电点溶解度提高,但中性p H下溶解度变小,且蛋白水解致使其表面电位绝对值明显提高;分离所得纤维聚集体的氨基酸组成和表面电位与热处理蛋白相似,在等电点和中性p H溶解度更低;多肽则含有较少疏水氨基酸和较多负电氨基酸,在p H 2.0~9.0溶解度较好,其在酸性p H下电荷量和静电排斥力较低,导致其以无定形聚集体的形式存在,而中性p H其电荷量较高导致多肽分子间相互作用减弱,聚集体解离。     

Influence of collector surface composition and water chemistry on the deposition of cerium dioxide nanoparticles: QCM-D and column experiment approaches

The deposition behavior of cerium dioxide (CeO(2)) nanoparticles (NPs) in dilute NaCl solutions was investigated as a function of collector surface composition, pH, ionic strength, and organic matter (OM). Sensors coated separately with silica, iron oxide, and alumina were applied in quartz crystal microbalance with dissipation (QCM-D) to examine the effect of these mineral phases on CeO(2) deposition in NaCl solution (1-200 mM). Frequency and dissipation shift followed the order: silica > iron oxide > alumina in 10 mM NaCl at pH 4.0. No significant deposition was observed at pH 6.0 and 8.5 on any of the tested sensors. However, ≥ 94.3% of CeO(2) NPs deposited onto Ottawa sand in columns in 10 mM NaCl at pH 6.0 and 8.5. The inconsistency in the different experimental approaches can be mainly attributed to NP aggregation, surface heterogeneity of Ottawa sand, and flow geometry. In QCM-D experiments, the deposition kinetics was found to be qualitatively consistent with the predictions based on the classical colloidal stability theory. The presence of low levels (1-6 mg/L) of Suwannee River humic acid, fulvic acid, alginate, citric acid, and carboxymethyl cellulose greatly enhanced the stability and mobility of CeO(2) NPs in 1 mM NaCl at pH 6.5. The poor correlation between the transport behavior and electrophoretic mobility of CeO(2) NPs implies that the electrosteric effect of OM was involved.     

Coprecipitation of arsenate with metal oxides: nature, mineralogy, and reactivity of aluminum precipitates

Arsenic mobilization in soils is mainly controlled by sorption/desorption processes, but arsenic also may be coprecipitated with aluminum and/or iron in natural environments. Although coprecipitation of arsenic with aluminum and iron oxides is an effective treatment process for arsenic removal from drinking water, the nature and reactivity of aluminum- or iron-arsenic coprecipitates has received little attention. We studied the mineralogy, chemical composition, and surface properties of aluminum-arsenate coprecipitates, as well as the sorption of phosphate on and the loss of arsenate from these precipitates. Aluminum-arsenate coprecipitates were synthesized at pH 4.0, 7.0, or 10.0 and As/Al molar ratio (R) of 0, 0.01, or 0.1 and were aged 30 or 210 d at 50 degrees C. In the absence of arsenate, gibbsite (pH 4.0 or 7.0) and bayerite (pH 10.0) formed, whereas in the presence of arsenate, very poorly crystalline precipitates formed. Short-range ordered materials (mainly poorly crystalline boehmite) formed at pH 4.0 (R = 0.01 and 0.1), 7.0, and 10.0 (R= 0.1) and did not transform into Al(OH)3 polymorphs even after prolonged aging. The surface properties and chemical composition of the aluminum precipitates were affected by the initial pH, R, and aging. Chemical dissolution of the samples by 6 mol L(-1) HCl and 0.2 mol L(-1) oxalic acid/ oxalate solution indicated that arsenate was present mainly in the short-range ordered precipitates. The sorption of phosphate onto the precipitates was influenced by the nature of the samples and the amounts of arsenate present in the precipitates. Large amounts of phosphate partially replaced arsenate only from the samples formed at R = 0.1. The quantities of arsenate desorbed from these coprecipitates by phosphate increased with increasing phosphate concentration, reaction time, and precipitate age butwere always lessthan 30% of the amounts of arsenate present in the materials and were particularly low (<4%) from the sample prepared at pH 4.0. Arsenate appeared to be occluded within the network of short-range ordered materials and/or sorbed onto the external surfaces of the precipitates, but sorption on the external surfaces seemed to increase by increasing pH of sample preparation and aging. Furthermore, at pH 4.0 more than in neutral or alkaline systems the formation of aluminum arsenate precipitates seemed to be favored. Finally, we have observed that greater amounts of phosphate were sorbed on an aluminum-arsenate coprecipitate than on a preformed aluminum oxide equilibrated with arsenate under the same conditions (R = 0.1, pH 7.0). In contrast, the opposite occurred for arsenate desorption, which was attributed to the larger amounts of arsenate occluded in the coprecipitate.     

Production of diarrheal toxin by selected strains of Bacillus cereus

The production of diarrheal toxin by six selected strains of Bacillus cereus was monitored during growth at 32 °C, a temperature described as near-optimal for growth and toxin production. Toxic activity was measured in culture filtrates and cellular extracts sampled at three different times during growth. Two alternative methods, a cytotoxicity test on Chinese hamster ovary (CHO) cells and a commercial immunological test (BCET-RPLA, Oxoid) were used. Toxin titres were in agreement with epidemiological characteristics and toxicity demonstrated by using other systems in other examinations. A comparison of intra- and extracellular toxicities measured at the exponential and stationary growth phases showed that the toxin was essentially secreted during the exponential phase. For several strains, secretion peaked during the period from the middle exponential phase to the beginning of the stationary phase. There was no important overall increase of the toxicity during full and late stationary phase. The level was stable or even lower, thus indicating that diarrheal toxin production during stationary phase was small, if any, and that the toxin was unstable under these conditions. Statistical analysis of toxicities showed that the cytotoxicity test was correlated with the immunological test (significant at a 1% level). For routine determinations, a toxicologic laboratory may use any of the two methods, depending oft its facilities, the immunological test being relatively expensive.     

Modeling the lag phase of Listeria monocytogenes

An estimate of the lag phase duration is an important component for predicting the growth of a bacterium and for creating process models and risk assessments. Most current research and data for predictive modeling programs initiated growth studies with cells grown to the stationary phase in a favorable pH, nutrient and temperature environment. In this work, Listeria monocytogenes Scott A cells were grown in brain heart infusion (BHI) broth at different temperatures from 4 to 37 degrees C to the exponential growth or stationary phases. Additional cells were suspended in a dilute broth, desiccated or frozen. These cells were then transferred to BHI broth at various temperatures from 4 to 37 degrees C and the lag phase durations were determined by enumerating cells at appropriate time intervals. Long lag phases were observed for cells initially grown at high temperatures and transferred to low temperatures. In general, exponential growth cells had the shortest lag phases, stationary phase and starved cells had longer, frozen cells had slightly longer and desiccated cells had the longest lag phases. These data were from immediate temperature transitions. When a computer-controlled water bath linearly changed the temperature from 37 to 5 degrees C over a 3.0- or 6.0-h period, the cells had short lags and grew continuously with declining growth rates. Transitions of 0.75 or 1.0 h had 20-h lag phases, essentially that of immediate transitions. When the transition was 1.5 h, an intermediate pattern of less than 1 log of growth followed by no additional growth for 20 h occurred.     

FLOCCULENCE OF BREWERY YEASTS AND THEIR SURFACE PROPERTIES: CHEMICAL COMPOSITION,ELECTROSTATIC CHARGE AND HYDROPHOBICITY

The flocculence of a top and a bottom fermentation brewery yeast was studied through the cell surface properties: surface composition (X-ray photoelectron spectroscopy), surface charge (microelectrophoresis) and hydrophobicity (adhesion tests). Aerobic Cultures and fermentation simulating industrial conditions were performed. The flocculence (tendency to flocculate in favourable conditions) has been determined by a test which separated the occurrence of flocculation and the measurement of its manifestation (suspension clearing); the flocculating agents were ethanol and calcium. The top fermentation strain does not flocculate with calcium alone but is very sensitive to ethanol (0.01% (v/v)). The lower flocculence observed for cells cultured on malt extract as compared with cells cultured on yeast extract is accompanied by a more negative zeta potential. The bottom fermentation strain which has a more negative surface flocculates only in the presence of calcium but ethanol enhances the process. The variation of flocculence according to the culture medium and harvesting time is correlated to the variation of the phosphate surface concentration. This is not paralleled by a variation of the zeta potential which does not decrease below a value of about-35 mV as the phosphate surface concentration increases. The increase of flocculence as a function of culture time correlates also with an increase of the cell hydrophobicity; this is more obvious for the cultures than for the fermentations.     

Determination of ovine casein heterogeneity using gel electrophoresis and immunochemical techniques.

Discontinuous PAGE at alkaline and acid pH, polyacrylamide gel isoelectric focusing, two dimensional electrophoresis and immunoblotting have been used to study the heterogeneity of sheep caseins. Three main phenotypes were selected either because of mobility at alkaline and acid pH of the individual alpha s components or because of their relative intensity. On electrophoresis at alkaline pH, one phenotype showed two distinct bands of lower electrophoretic mobility than beta 1- and beta 2-caseins. A detailed study of these components using immunospecific detection with beta-casein antiserum showed that these minor components of ovine casein may have a polypeptide chain similar to that of beta 1- and beta 2-caseins. Complete electrophoretic patterns of the casein components in some individual milks are also presented.