Guillaume Delaplace

Influence of calcium on β-lactoglobulin denaturation kinetics: Implications in unfolding and aggregation mechanisms

Much research dealing with the processing of milk by-products in heat exchangers has noted the key role of calcium in β-lactoglobulin (β-LG) fouling behavior. Nevertheless, the manner by which Ca affects β-LG denaturation has rarely been quantified using reliable kinetic and thermodynamic data. To this end, the influence of Ca on β-LG denaturation mechanisms in simulated lactoserum concentrates was studied on the laboratory-scale under 100°C by HPLC analysis. The heat-treated solutions were composed of 53.3g/L β-LG and were enriched in Ca at various concentrations (0, 66, 132, and 264 mg/kg). The kinetic parameters (reaction order, activation energy, and frequency factor) associated with β-LG denaturation, along with the unfolding and aggregation thermodynamic parameters were deduced from these experiments and discussed with respect to Ca content. We found that the multistage process characterizing β-LG thermal denaturation is not greatly affected by Ca addition. In fact, the general model subdividing β-LG denaturation mechanisms in 2 steps, namely, unfolding and aggregation, remained valid for all tested Ca concentrations. The change in the predominant mechanism from unfolding to aggregation was observed at 80°C across the entire Ca concentration range. Moreover, the classical 1.5 reaction order value was unaffected by the presence of Ca. Interpretation of the acquired kinetic data showed that Ca addition led to a significant increase in kinetic rate, and more so in the aggregation temperature range. This indicates that Ca principally catalyzes β-LG aggregation, by lowering the Coulombian repulsion between the negatively charged β-LG reactive species, bridging β-LG proteins, or via an ion-specific conformational change. To a lesser extent, Ca favors β-LG unfolding, probably by disturbing the noncovalent binding network of native β-LG. Simultaneously, Ca has a slight protective role on the native and unfolded β-LG species, as shown by the increase in activation energy with Ca concentration. The calculation of thermodynamic parameters related to β-LG denaturation confirmed this observation. A threshold effect in Ca influence was noted in this study: no further significant kinetic rate change was observed above 132 mg/kg of Ca; at this concentration, the studied solution was an almost equimolar mixture of β-LG and Ca. Finally, we simulated the temporal evolution of β-LG species concentrations at diverse Ca contents at 3 holding temperatures. The simulations were based on the acquired kinetic parameters. This permitted us to highlight the greater effect of Ca on β-LG denaturation at high Ca content or for short-time heat treatments at temperatures near 100°C, as in heat exchangers.

Toward the understanding of the interfacial dairy fouling deposition and growth mechanisms at a stainless steel surface: a multiscale approach.

The microstructures of two dairy fouling deposits obtained at a stainless steel surface after different processing times in a pilot plate heat exchanger were investigated at different scales. Electron-Probe Micro Analysis, Time-of-Flight Secondary Ion Mass Spectrometry, Atomic Force Microscopy, and X-Ray Photo-electron Spectroscopy techniques were used for this purpose. The two model fouling solutions were made by rehydrating whey protein in water containing calcium or not. Results on samples collected after 2h processing show that the microstructure of the fouling layers is completely different depending on calcium content: the layer is thin, smooth, and homogeneous in absence of calcium and on the contrary very thick and rough in presence of calcium. Analyses on substrates submitted to 1 min fouling reveal that fouling mechanisms are initiated by the deposit of unfolded proteins on the substrate and start immediately till the first seconds of exposure with no lag time. In presence of calcium, amorphous calcium carbonate nuclei are detected in addition to unfolded proteins at the interface, and it is shown that the protein precedes the deposit of calcium on the substrate. Moreover, it is evidenced that amorphous calcium carbonate particles are stabilized by the unfolded protein. They are thus more easily trapped in the steel roughnesses and contribute to accelerate the deposit buildup, offering due to their larger characteristic dimension more roughness and favorable conditions for the subsequent unfolded protein to depose.

Experimental and CFD Simulation of Heat Transfer to Highly Viscous Fluids in an Agitated Vessel Equipped With a non Standard Helical Ribbon Impeller

In this work, heat transfer has been investigated for several Newtonian and non-Newtonian liquids agitated in a rounded bottom vessel equipped with an atypical helical ribbon impeller. Both experimental and numerical approaches were attempted. The measured rates of heat transfer from the wall of the vessel to the agitated liquid allow the comparison of heat transfer performances of this mixing system with those obtained on standard helical ribbon impeller. A method was indicated to correlate non-Newtonian results with those established with Newtonian liquids. No temperature gradient was detected in the bulk. Numerical approach allows the local heat transfer coefficient to be obtained, and underlines the difficulty to relate global heat transfer coefficient to the thickness of the thermal boundary layer.

Laminar Mixing Performances of a Stirred Tank Equipped with Helical Ribbon Agitator Subjected to Steady and Unsteady Rotational Speed

This paper investigates the way of improving the mixing of highly viscous Newtonian fluids in a tank using suitable operating conditions (unsteady stirring approaches). The agitator used is a non standard helical ribbon impeller fitted with an anchor at the bottom. The degree of homogeneity in the tank is followed using a conductivity method after a tracer injection. It is shown that the use of time-dependent rotational speed during the mixing process allows energy savings. For the unsteady stirring approaches tested, energy savings can reach up to 60% compared to the energy required to obtain the same mixing time with constant impeller rotational speed. Finally, a model is provided which allows the prediction of the mixing process of the agitated system with Newtonian fluids for both steady and unsteady stirring approaches.

Power requirement when mixing a shear-thickening fluid with a helical ribbon impeller type

The optimal design of close clearance impellers requires the knowledge of the power demand of the mixing equipment. In non-Newtonian mixing, this can be readily obtained using the Metzner and Otto concept. In this work, this concept and the determination of the K s (constant of proportionality which links the effective shear rate (defined by the Metzner and Otto concept) and the rotational speed of the impeller when mixing non-Newtonian fluids under laminar regime) value for an atypical helical agitator (PARAVISC system from Ekato firm) have been revised in the case of shear-thinning fluids and a shear-thickening fluid. For poor shear-thinning fluids, it has been shown that for the mixing system the K s value does not vary strongly with the flow behavior index, and may be regarded as a constant for the mixing purpose design. By contrast, for the shear-thickening fluid, power consumption measurements indicate that the relationship between the K s values and the flow behavior index is much more complex due to a partial solidification of the product around the impeller.

Predicting the distribution of whey protein fouling in a plate heat exchanger using the kinetic parameters of the thermal denaturation reaction of β-lactoglobulin and the bulk temperature profiles

Fouling of plate heat exchangers (PHE) is a severe problem in the dairy industry, notably because the relationship between the build-up of protein fouling deposits and the chemical reactions taking place in the fouling solution has not yet been fully elucidated. Experiments were conducted at pilot scale in a corrugated PHE, and fouling deposits were generated using a model β-lactoglobulin (β-LG) fouling solution for which the β-LG thermal denaturation reaction constants had been previously determined experimentally. Then 18 different bulk temperature profiles within the PHE were imposed. Analysis of the fouling runs shows that the dry deposit mass per channel versus the ratio R=kunf/kagg (with kunf and kagg representing, respectively, the unfolding and aggregation rate constants computed from both the identification of the β-LG thermal denaturation process and knowledge of the imposed bulk temperature profile into the PHE channel) is able to gather reasonably well the experimental fouling mass data into a unique master curve. This type of representation of the results clearly shows that the heat-induced reactions (unfolding and aggregation) of the various β-LG molecular species in the bulk fluid are essential to capture the trend of the fouling mass distribution inside a PHE. This investigation also illustrates unambiguously that the release of the unfolded β-LG (also called β-LG molten globule) within the bulk fluid (and the absence of its consumption in the form of aggregates) is a key phenomenon that controls the extent of protein fouling as well as its location inside the PHE.

Denaturation Kinetics of Whey Protein Isolate Solutions and Fouling Mass Distribution in a Plate Heat Exchanger

Few investigations have attempted to connect the mechanism of dairy fouling to the chemical reaction of denaturation (unfolding and aggregation) occurring in the bulk. The objective of this study is to contribute to this aspect in order to propose innovative controls to limit fouling deposit formation. Experimental investigations have been carried out to observe the relationship between the deposit mass distribution generated in plate heat exchangers (PHE) by a whey protein isolate (WPI) mainly composed of β-lactoglobulin (β-Lg) and the ratio between the unfolding and aggregation rate constants. Experiments using a PHE were carried out at a pilot scale to identify the deposit distribution of a model fouling solution with different calcium contents. In parallel, laboratory experiments were performed to determine the unfolding/aggregation rate constants. Data analysis showed that (i) β-Lg denaturation is highly dependent on the calcium content, (ii) for each fouling solution, irrespective of the imposed temperature profile, the deposit mass in each channel and the ratio between the unfolding and aggregation rate constants seem to be well correlated. This study demonstrates that both the knowledge of the thermal profile and the β-Lg denaturation rate constants are required in order to predict accurately the deposit distribution along the PHE.

Advances in food powder agglomeration engineering.

Food powders are used in everyday life in many ways and offer technological solutions to the problem of food production. The natural origin of food powders, diversity in their chemical composition, variability of the raw materials, heterogeneity of the native structures, and physicochemical reactivity under hydrothermal stresses contribute to the complexity in their behavior. Food powder agglomeration has recently been considered according to a multiscale approach, which is followed in the chapter layout: (i) at the particle scale, by a presentation of particle properties and surface reactivity in connection with the agglomeration mechanisms, (ii) at the mechanisms scale, by describing the structuration dynamics of agglomerates, (iii) at the process scale, by a presentation of agglomeration technologies and sensors and by studying the stress transmission mode in the powder bed, and finally (iv) by an integration of the acquired knowledge, thanks to a dimensional analysis carried out at each scale.

Antifouling Stainless Steel Surface: Competition between Roughness and Surface Energy

To increase the shelf-life qualities of dairy products, a heat treatment is usually done. However, heat treatments induce physico-chemical modifications of the products. Some of them lead to the expected product but an unwanted consequence of this process is the formation of a fouling deposit on the surfaces in contact with the processed fluid. To eliminate fouling, cleaning processes have to be done once a day. It increases the processing and maintenance costs. To control and to decrease the fouling are the main problems in food industries and an active research is carried out on efficient antifouling surface treatments. In the present study, a 316L 2B stainless steel was submitted to different surface treatments (Flame and plasma pre-treatments, Plasma Enhanced Chemical Vapour Deposition, hydrophobic coatings, mechanical polishing ...) to try to establish correlations between different surface parameters (roughness, hydrophobicity, nanostructuration, surface energy, ...) onto the fouling in heat exchangers. All the treated plates were then submitted to a fouling test using an aqueous solution of β-lactoglobulin at 1% (p/p) with a final calcium concentration of 910 mg/L and compared to a bare steel plate. The results obtained imply different influences of each parameter depending on the surface roughness: the effect of a non organized micrometric roughness is preponderant compared to the surface energy: the fouling comes from a mechanical effect mainly due to rubbing. However, when the surface is nanostructured, fouling decreases. When the roughness reaches the nanometer scale (between 100 and 400 nm), it is the surface energy and the polar/apolar components which become preponderant compared to the roughness. Fouling is this time mainly due to the hydrophilicity of the surface and to the adsorption of the β-lactoglobulin on acido-basic sites. Finally, when the roughness reaches less than 50 nm, polar/apolar components show no effect anymore, the preponderant parameter is the hydrophobicity of the surface.

Antifouling Biomimetic Liquid-infused Stainless Steel: Application to Dairy Industrial Processing.

Fouling is a widespread and costly issue, faced by all food-processing industries. Particularly, in the dairy sector, where thermal treatments are mandatory to ensure product safety, heat-induced fouling represents up to 80% of the total production costs. Significant environmental impacts, due the massive consumption of water and energy, are also to deplore. Fouling control solutions are thus desperately needed, as they would lead to substantial financial gains as well as tremendous progress toward eco-responsible processes. This work aims at presenting a novel and very promising dairy fouling-mitigation strategy, inspired by nature, and to test its antifouling performances in real industrial conditions. Slippery liquid-infused surfaces were successfully designed directly on food grade stainless steel, via femtosecond laser ablation, followed by fluorosilanization and impregnation with an inert perfluorinated oil. Resulting hydrophobic surfaces (water contact angle of 112°) exhibited an extremely slippery nature (contact angle hysteresis of 0.6°). Outstanding fouling-release performances were obtained for these liquid-infused surfaces as absolutely no trace of dairy deposit was found after 90 min of pasteurization test in pilot-scale equipment followed by a short water rinse.