Protein hydration, interfacial

During ageing of the mix, interfacial milk protein hydration also increases simultaneously with protein desorption from the fat globules. The water content of the isolated cream layers after centrifugation of ice cream mix can be analyzed by Karl Fischer titration. From such analyses, interfacial protein hydration can be calculated (Figure 13). 

The voluminosity or hydration of interfacially bound protein may be calculated from the amount of water bound per gram of fat divided by the amount of protein bound per gram of fat. This corresponds to the volume of water per gram interfacial protein. Calculations show that emulsifiers facilitate interfacial protein hydration. This property is probably connected with their ability to desorb protein from the interface (Figure 14). 

The second section. Perspective Macro- and Microinteractions of Water and Polymers, consists of three chapters. It covers a brief historical introduction leading to an insight into selected aspects of current thought on interactions of water with model solutes and proteins [4], consideration of water-polymer and ice-polymer interfacial regions [5], and examination of stages in the process of protein hydration assessed by a variety of types of measurements [6]. 

D. E. Khoshtariya, E. Hansen, R. Leecharoen, and G. C. Walker, Probing protein hydration by the difference 0-H (O-D) vibrational spectroscopy Interfacial percolation network involving highly polarizable water-water hydrogen bonds. J. Mol. Liq., 105 (2003), 13-36. 

The NMR study by Wiithrich and coworkers has shown that there is a cavity between the protein and the DNA in the major groove of the Antennapedia complex. There are several water molecules in this cavity with a residence time with respect to exchange with bulk water in the millisecond to nanosecond range. These observations indicate that at least some of the specific protein-DNA interactions are short-lived and mediated by water molecules. In particular, the interactions between DNA and the highly conserved Gin 50 and the invariant Asn 51 are best rationalized as a fluctuating network of weak-bonding interactions involving interfacial hydration water molecules. 

The new generations of experiments are aimed at linking dynamical studies of these and other processes to the function. We have already begun research in this direction. In a recent publication [9] we reported studies of the femtosecond dynamics of an RNA-protein complex and then compared the results with those obtained for in vivo (E. Coli) transcription anti-termination activities. In two other studies we measured the activity of the protein Subtilisin Carlsberg, discussed above, to a substrate, and the role of hydration in interfacial binding and function of bovine pancreatic phospholipase at a substrate site. The goal in all these studies is to relate structures to the dynamics and hopefully to key features of the (complex ) function. 

It is commonly stated that the first readily observable event at the interface between a material and a biological Quid is protein or macromolecule adsorption. Clearly other interactions precede protein adsorption water adsorption and possibly absorption (hydration effects), ion bonding and electrical double layer formation, and the adsorption and absorption of low molecular weight solutes — such as amino acids. The protein adsorption event must result in major perturbation of the interfacial boundary layer which initially consists of water, ions, and other solutes. 

Figure 14 Effect of low temperature on hydration of bovine casein micelles and of interfacially bound protein in ice cream mix with (+E) and without (-E) emulsifier (saturated mono-diglyceride).

The properties of the water phase in whippable emulsions are important for product stability. The water phase is influenced by the soluble components of the systems, i.e., sugars, proteins and hydrocolloids. Interfacial hydration may also influence the properties of the water phase, particularly in high-fat systems. 

Both hydrocolloids and emulsifiers increase the water-binding capacity in the mix (increased % of hydrogen atoms with low T2 and decreased T2 values). A synergistic effect is observed when both ingredients are present. From studies described earlier in this chapter, the effect of hydrocolloids is assumed to be due to simple water binding and increased thickness of protein layers around the fat globules, whereas the effect of emulsifiers may be due to the increased hydration of interfacially bound protein as well as increased hydration of polar groups of emulsifier at the oil-water interface. 

Protein function at solid-liquid interfaces holds a structural and a dynamic perspective [31]. The structural perspective addresses macroscopic adsorption, molecular interactions between the protein and the surface, collective interactions between the individual adsorbed protein molecules, and changes in the conformational and hydration states of the protein molecules induced by these physical interactions. Interactions caused by protein adsorption are mostly non-covalent but strong enough to cause drastic functional transformations. All these features are, moreover, affected by the double layer and the electrode potential at electrochemical interfaces. Factors that determine protein adsorption patterns have been discussed in detail recently, both in the broad context of solute proteins at solid surfaces [31], and in specific contexts of interfacial metalloprotein electrochemistry [34]. Some important elements that can also be modelled in suitable detail would be ... 

In the bacterial PI-PLC structures, the top of the barrel rim has several hydrophobic residues that are fully exposed to solvent and poorly defined in the crystal structures (implying significant mobility). The active site of PI-PLC is accessible and well-hydrated, and these mobile elements at the top of the barrel offer a different motif for interactions of the protein with phospholipid interfaces. The PI-PLC from B. thuringiensis (nearly identical in sequence to the enzyme from B. cereus whose crystal structure was determined) exhibits the property of interfacial activation, where enhanced activity is observed when the substrate PI is present in an interface compared to monomeric substrate (Lewis et al., 1993). However, other non-substrate lipids such as phosphatidylcholine (PC), phosphatidic acid (PA), and other anionic lipids have an effect on the activity of PI-PLC toward both substrates PI and water-soluble cIP (Zhou et al., 1997). In particular, the presence of PC enhances the catalytic activity of... 

Moreover, in the case of the low level of hydration h = 0.25 the evolution of the density of states as a function of the temperature is less pronounced than in the case of h = 0.5. This is in agreement with the structural study [43] at the lower hydration (h = 0A 75), which only detected small changes when the temperature is lowered from room temperature to 77 K, and with further structural studies of low-hydrated Vycor samples. Low temperatures do not significantly affect the overall structure of the protein and the bound water molecule, and no crystallization of water has been observed. This could reflect the fact that at room temperature the interfacial water behaves like a dense supercooled liquid. 

Steric Hindrance. Another form of stabilization is relatively independent of ionic strength the oil droplets are prevented from making contact by simple steric hindrance. This may take two forms, either an immobilized water layer at the interface or a solid interfacial film. Emulsion stabilization by proteins, gums, and polyoxyethylene derivatives occurs by the first mechanism. Hydrophobic parts of the stabilizers adsorb at the oil surface, but adjacent large hydrophilic segments are hydrated and form an immobilized layer on the order of 10-100 nm thick (Figure 9). As mentioned, these hydrated segments frequently interact to cause flocculation, while coalescence of the oil drops themselves is prevented. Such emulsions are frequently used as carriers for oil-soluble flavors, essences, and colorants. 

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