Effect of pH change on size and nanomechanical behavior of whey protein microgels.

Microgels specific structural and functional features are attracting high research interest in several applications such as bioactives and drug delivery or functional food ingredients. Whey protein microgels (WPM) are obtained by heat treatment of whey protein isolate (WPI) in order to promote intramolecular cross-linking. In the present work, atomic force microscopy (AFM) was used in contact mode and in liquid to investigate WPM particles topography and mechanical properties at the nanoscale at native pH (6.5) and acid pH (5.5 and 3.0). Prior to AFM, WPM particles were captured on a gold substrate via low energy interactions by means of specific monoclonal antibodies. AFM images clearly showed an increase in the size of WPM particles induced by pH decrease. AFM in force spectroscopy mode was employed to monitor the elasticity of WPMs. The obtained effective Young's modulus data showed a significant increase in stiffness at pH 5.5 and pH 3.0, over 15-fold compared to native pH. These findings indicate that the mechanical profile of the WPM network varied with the pH decrease. The WPM topographic and nanomechanical changes induced by acidification were most likely due to substantial changes in the shape and inner structure of WPM particles. Our results suggest that internally cross-linked structures, modified by acidification could display interesting functional properties when used as a food ingredient.

Binding analysis between monomeric ß-casein and hydrophobic bioactive compounds investigated by surface plasmon resonance and fluorescence spectroscopy.

ß-Casein, a phosphoprotein representing 37% of the bovine milk caseins, has specific features promoting its application as a nanocarrier for hydrophobic bioactives. In this study, the interactions of ß-casein with curcumin and vitamin D3 under the same physico-chemical conditions were investigated. The interaction kinetics have been studied by surface plasmon resonance (SPR) and fluorescence spectroscopy. The KD value for curcumin-ß-casein interaction has been successfully evaluated (4.1 ±â€¯0.7 × 10-4 M) using SPR by fitting data to a 1:1 Langmuir interaction model. Conversely, the SPR responses obtained for vitamin D3 show that the interactions between this hydrophobic compound and the ß-casein immobilized on the sensor chip were below the sensitivity of the SPR apparatus. Moreover, the fluorescence quenching data show that curcumin has higher affinity to ß-casein (KA = 23.5 ±â€¯1.9 × 104 M-1) than vitamin D3 (KA = 5.8 ±â€¯1.1 × 104 M-1).

Atomic Force Microscopy Study of the Topography and Nanomechanics of Casein Micelles Captured by an Antibody.

Casein micelles (CMs) are colloidal phospho-protein-mineral complexes naturally present in milk. This study used atomic force microscopy (AFM) in a liquid environment to evaluate the topography and nanomechanics of single native CMs immobilized by a novel capture method. The proposed immobilization method involves weak interactions with the antiphospho-Ser/Thr/Tyr monoclonal antibody covalently bound to a carboxylic acid self-assembled monolayer (SAM) on a gold surface. This capture strategy was compared to the commonly used covalent immobilization method of CMs via carbodiimide chemistry. With this conventional method, CMs remained mainly mobile during AFM measurements in liquid, disturbing the evaluation of their average size and elastic properties. Conversely, when captured by the specific antibody, they were successfully immobilized and their integrity was preserved during the AFM measurement. The characterization of both CM topography and elastic properties was carried out in a liquid ionic environment at native pH 6.6. The CMs' capture efficiency via antibody was concurrently proved by surface plasmon resonance. The calculation of casein micelles' width, height, and contact angle was carried out from the recorded 2D AFM images. CMs were characterized by a mean width of 148 ± 8 nm and a mean height of 42 ± 1 nm. Weak forces were applied to single captured CMs. The obtained force versus indentation curves were fitted using the Hertz model in order to evaluate their elastic properties. The elasticity distribution of native CMs exhibited a unimodal trend with a peak centered at 269 ± 14 kPa.

Production, Characterization, and Application of Bacillus licheniformis W16 Biosurfactant in Enhancing Oil Recovery.

The biosurfactant production by Bacillus licheniformis W16 and evaluation of biosurfactant based enhanced oil recovery (EOR) using core-flood under reservoir conditions were investigated. Previously reported nine different production media were screened for biosurfactant production, and two were further optimized with different carbon sources (glucose, sucrose, starch, cane molasses, or date molasses), as well as the strain was screened for biosurfactant production during the growth in different media. The biosurfactant reduced the surface tension and interfacial tension to 24.33 ± 0.57 mN m-1 and 2.47 ± 0.32 mN m-1 respectively within 72 h, at 40°C, and also altered the wettability of a hydrophobic surface by changing the contact angle from 55.67 ± 1.6 to 19.54°± 0.96°. The critical micelle dilution values of 4X were observed. The biosurfactants were characterized by different analytical techniques and identified as lipopeptide, similar to lichenysin-A. The biosurfactant was stable over wide range of extreme environmental conditions. The core flood experiments showed that the biosurfactant was able to enhance the oil recovery by 24-26% over residual oil saturation (Sor). The results highlight the potential application of lipopeptide biosurfactant in wettability alteration and microbial EOR processes.