Thermal isoelectric precipitation of alpha-lactalbumin from a whey protein concentrate: Influence of protein-calcium complexation

The selective precipitation of alpha-lactalbumin (alpha-LA) at a pH around its isoelectric point (4.2) under heat treatment is the basis for a fractionation process of whey proteins. As precipitation is a phenomenon dependent on the protein hydrophobicity, and as the release of the tightly bound calcium occurring at pH around 4 modifies the alpha-LA hydrophobicity, the specific role of calcium on isoelectric precipitation is investigated. A study of the extent of alpha-LA precipitation in a whey protein concentrate under various operating conditions of pH, temperature, protein concentration, and calcium content is presented. We propose a mechanism for this phenomenon as a combination of a complexation equilibrium and of an irreversible precipitation, to account for the influence of temperature, alpha-LA concentration total ionic content, and calcium concentration, and also to estimate the complexation equilibrium constant. (c) 1995 John Wiley & Sons, Inc.     

An analysis of proton fluxes coupled to electron transport and ATP synthesis in chloroplast thylakoids

Electron transport, phosphorylation and internal proton concentration were measured in illuminated spinach chloroplast thylakoid membranes under a number of conditions. Regardless of the procedure used to vary these parameters, the data fit a simple chemiosmotic model. Protons from Photosystem II did not appear to be utilized differently from those derived from Photosystem I. The maximal phosphorylation efficiency ($\text{P}\text{e}{}_2$) for photophosphorylation in washed thylakoids under oxidizing conditions is likely to be $\text{4}\text{3}$. This value is consistent with a proton-to-electron-pair ratio of 4 for electron flow through both photosystems and a proton-to-ATP ratio of 3 for the chloroplast proton-ATPase.     

Precipitation of collagens by polyethylene glycols

Types I, II, and III collagens are readily precipitated at neutral pH by polyethylene glycols (PEG). As the molecular weight fraction of the polyethylene glycols increases, they become more effective as precipitants on a weight basis. The amount of PEG required for precipitation depends on the pH, the ionic strength, and the nature of the buffer or salts present. In tissue culture media, low concentrations of collagens and procollagens may be quantitatively precipitated and readily collected by low-speed centrifugation. Polyethylene glycol precipitation can be used to obtain collagens and procollagens from tissue culture media at either analytical or preparative scale, and since the polyethylene glycols do not bind to collagens, the precipitates may be further analyzed directly by chromatographic or electrophoretic methods.     

Molecular heterogeneity of canine cholecystokinin in portal and peripheral plasma

The release of molecular forms of cholecystokinin (CCK) into the portal and peripheral blood in response to an intraduodenal perfusion of sodium oleate (9 mmol X h-1) was studied in six conscious dogs with chronic portal vein catheters. Immunoreactive CCK as concentrated from 20 ml plasma by C18 SEP PAK cartridges and the pattern of molecular forms of CCK were studied by G50 gel filtration. CCK-like immunoreactivity (CCK-LI) was measured in the column eluates with antibody 5135, which measures gastrin and CCK equally and requires the intact carboxyl-terminus for full recognition. Gastrin was measured specifically with antibody 1611. Intraduodenal perfusion with oleate did not alter basal gastrin release. Release of CCK-LI by intraduodenal oleate was calculated by the increments of the integrated CCK-LI peaks over basal. Total CCK-like immunoreactivity (CCK-LI), calculated by integration of all CCK-LI peaks in gel filtration eluates, increased over basal by 12 fmol/ml in the portal and by 6 fmol/ml in the peripheral plasma after intraduodenal perfusion with sodium oleate. The main molecular forms eluted on gel filtration in positions of CCK33,39 and of CCK8. The pattern of CCK in the peripheral plasma was similar to that in the portal plasma except that in the peripheral plasma large molecular forms were more abundant than small forms. This finding was confirmed when CCK39 and CCK8 were infused either into the portal vein or into the peripheral vein and peripheral plasma CCK levels were measured. Elimination of CCK8 after portal vein infusion compared to peripheral vein infusion was about 3 times higher than that of CCK39. The abundance of large molecular forms of CCK in the circulating blood which are similar in potency to small forms, underlines their role in the physiology of CCK.     

Purification of recombinant protein A by aqueous two-phase extraction integrated with affinity precipitation

Aqueous two-phase extraction incorporated affinity precipitation was examined as a technique for protein purification. An enteric coating polymer, Eudragit S100, was employed as a ligand carrier. Eudragit was specifically partitioned to the top phase in the aqueous two-phase systems. For application of this method to purification of recombinant protein A using human IgG coupled to Eudragit in an aqueous two-phase system, 80% of protein A added was recovered with 81% purity. The purity was enhanced 26-fold by thid method. The IgG-Eudragit could be used repeatedly for the purification process. This seperation method should be applicable to industrial-scale purification as a new purification procedure combining the advantages and compensating for the disadvantages of the aqueous two-phase method and affinity precipitation method. (c) 1992 John Wiley & Sons, Inc.     

Carbon isotope discrimination at vegetative stage as an indicator of yield and water use efficiency of spring wheat (Triticum turgidum L. var. durum)

A field experiment was conducted to investigate if carbon isotope (¹³C) discrimination () measured at the vegetative stage of spring wheat (Triticum turgidum L. var. durum) is related with the yield and water use efficiency (WUE) at ripening. A line source sprinkler irrigation system exposed the wheat genotypes to different watering regimes, from rainfed to full irrigation and thereby increased the range in yield and WUE attainable in the four genotypes studied. The results indicated that values measured at the late stem elongation stage 60 days after planting (DAP), showed strong positive correlation with total dry matter yield (r=0.732^***), and a highly significant negative correlation with WUE (r=–0.755^***) measured at ripening 105 DAP. The data suggest that the imprints of measured at vegetative growth stage persists throughout the entire growth period, until maturity. Subject to confirmation from additional studies in other crops and locations, early measurements of may prove a useful tool for rapid and early screening of cultivars, for high yield and high WUE.     

Evaluation of actual evapotranspiration of a Quercus ilex L. stand by the Bowen Ratio-Energy Budget method

Actual evapotranspiration from a closed-canopy Quercus ilex L. stand has been estimated by applying the Bowen Ratio-Energy Budget method. Daily water loss was 3.5 mm day^–1, with a peak rate near 0.6 mm hour^–1. The phenomenon of thermal inversion, quite common in mediterranean climates, seemed to play a significant role in reducing evapotranspiration, by promoting dew formation and delaying the establishment of fluxes of latent and sensible heat away from the canopy. Dew, which may form over many hours in the night, appears to be a major sink of available energy in the early morning and may represent a useful water source for stressed foliage. The alternating processes of condensation and evaporation may have a beneficial effect on the closed stand micro-environment.     

The evolution of saline lake waters: gradual and rapid biogeochemical pathways in the Basotu Lake District,Tanzania

The biogeochemical evolution of solutes markedly alters the chemistry in the closed-basin maar lakes that comprise the Basotu Lake District (Tanzania, East Africa). Examination of 11 (out of 13) lakes in the Basotu Lake District identified two distinct evolutionary pathways: a gradual path and a rapid path. During the course of biogeochemical evolution these waters follow either the gradual path alone or a combination of the gradual and rapid paths. Solute evolution along the gradual path is determined by all of the biogeochemical processes that for these waters appear to be tightly coupled to evaporative concentration (e.g. mineral precipitation, sorption and ion exchange, C0₂ degassing, and sulfate reduction). Rapid evolution occurs when mixing events suddenly permit H₂S to be lost to the atmosphere. The chemistry of waters undergoing rapid evolution is changed abruptly because loss of every equivalent of sulfide produces an equivalent permanent alkalinity.The Basotu Lake District in north central Tanzania is comprised of 13 maar lakes. They range in surface water conductivity from 592 to 24 000 µ S cm ^–1 (at 20°). Within these lake basins only a few of the variety of geo- and biogeochemical processes known to occur in lakes of this type are actually responsible for the gain and/or loss of individual solutes. For example, potassium appears to be taken up in the formation of illite. Calcium is precipitated as calcite. Magnesium interacts with alumino-silicate precursors to form a variety of clay minerals that contain magnesium (e.g. stevensite). This process is also known as reverse weathering. Sulfate is reduced to sulfide and subsequently lost as H₂S and/or metal sulfides. Alkalinity is lost owing to calcite precipitation and as a consequence of reverse weathering. Alkalinity is gained in the form of extra permanent alkalinity when sulfide is lost from these waters (via metal sulfide precipitation or gaseous emission to the atmosphere). Rapid (punctuated) evolution can occur in any lake containing anoxic waters providing that mixing events take place which cause H₂S to be lost to the atmosphere.Peter Kilham died on March 20, 1989, in Kisumu, Kenya, while working as part of a research team on Lake Victoria.     

The pH-independent effect of aluminum on cultures of phytoplankton from an acidic Wisconsin lake

Negative correlations between aluminum and planktonic algal abundance have been reported in acidic lakes. Natural assemblages of phytoplankton from a low-pH, low-Al lake (Franklin Lake, WI) were grown in semi-continuous cultures consisting of four treatments at pH 5.7 with 0.0, 50, 100, and 200 μg Al L⁻¹ and one treatment at pH 4.7 with no Al added. Asterionella ralfsii var. americana (a common diatom plankter in acidic lakes) grew well at both pH 4.7 and 5.7 when no Al was added but declined in all other treatments and so may be useful as an indicator of acidic, low monomeric-Al conditions. Other common plankters that showed this pattern included: Arthrodesmus indentatus, Ar. octocornus, Ar. quiriferus, Staurastrum arachne var. curvatum, S. longipes var. contractum, and S. pentacerum. Common taxa showing no toxic effects of Al were Dinobryon bavaricum, Peridinium limbatum, Stenokalyx monilifera, Elaktothrix sp. and Oedogonium sp. We hypothesize that metal toxicity as a pulse at spring snowmelt could dramatically change algal succession in moderately acidic lakes. The experimental results agreed well with field observations. These types of experiments are useful for predicting the responses of natural phytoplankton communities to increases in Al concentration.     

Effects of acid irrigation and liming on two clones of Norway spruce

Summary The effect of acid irrigation on the growth of rooted cuttings ofPicea abies (L.) Karst, was investigated in a pot experiment lasting 3 years. It involved two clones of Norway spruce, H 253 Bogstad I and H 254 Bogstad II. Irrigation water of pH 5.4, 4.0, 3.0 and 2.5 was used. Liming was included in the experiment.After the experimental period, the plants of all treatments were growing reasonably well. However, those plants irrigated at pH 2.5 were slightly discoloured. The plant mortality was only 3% throughout the experiment, and was not connected to acid irrigation. The limiting growth factor was N. All other nutrient elements measured in the plants were close to optimal concentration. Plants irrigated at pH 2.5, and to some extent at pH 3, contained excessively high concentrations of Al, t-S and SO₄. The total amount of Ca, Fe and Mn taken up by the plants decreased with increasing soil acidity. The increased growth of clone H 254 relative to H 253, produces a corresponding impression on soil characteristics. Soil acidity is governed by acid irrigation and CaCO₃ application, but the clonal effects are also of importance. Norway spruce appears to be tolerant to Al concentrations as high as 50 mmol/kg in the needles.