DIFFERENTIAL STABILIZING EFFECTS OF BUFFERS ON STRUCTURAL STABILITY OF BOVINE SERUM ALBUMIN AGAINST UREA DENATURATION

The conformational stability of bovine serum albumin (BSA) against urea denaturation was investigated in aqueous solutions both in the absence and presence of buffers. Various buffers differing in polar and nonpolar characters such as sodium phosphate, Tris-HCl, (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) HEPES and [3-(N-morpholino)propanesulfonic acid] MOPS buffers were used in this study. Urea-induced structural changes were analyzed using different probes, i.e., intrinsic fluorescence, ANS fluorescence and UV-difference spectral signal. Presence of different buffers in the incubation medium offered different degrees of resistance to the protein against urea-induced structural changes compared to those obtained in water (in the absence of buffers). A similar trend of buffer-induced structural resistance was noticed with three different probes. The stabilizing effect of these buffers followed the order: MOPS > HEPES > sodium phosphate > Tris-HCl > water. As found in MOPS and HEPES buffers, the highest stability of BSA can be attributed to the presence of morpholine and piperazine rings, respectively, in their structures. These groups might have produced a hydrophobic environment around the protein surface, thus stabilizing protein conformation against urea denaturation.

Synthesis and Characterization of an Amphoteric Asphalt Emulsifier

A novel amphoteric asphalt emulsifier of octadecylbis(propanamide)-(3’-sodium phosphate-2’-hydroxypropyl)ammonium chloride was synthesised from the raw materials octadecylamine, acrylamide, epichlorohydrin and sodium dihydrogen phosphate. The tertiary amine octadecyl-bis(propanamide) was synthesised from octadecylamine and acrylamide (step 1). Sodium 3-chloro-2-hydroxypropyl phosphate (intermediate) was obtained from epichlorohydrin and sodium dihydrogen phosphate (step 2). The asphalt emulsifier was obtained from octadecyl-bis(propanamide)-tertiary amine and the intermediate by quaternisation reaction (step 3). The yield of the final product reached 94.90%. The structure was identified by FTIR, 1H-NMR and elemental analysis. The critical micelle concentration of the product is 1.46 × 10–5 mol L–1. The surface tension at CMC is 37.78 mN ν–1. The saturated adsorption amount of asphalt emulsifier is 2.72 × 10–3 mmol ν–2. The occupied area per asphalt emulsifier molecule at CMC is 0.611 nm2 mol–1. The surfactant is a fast-setting asphalt emulsifier.