Hyperconcentrated Sweet Whey, a New Culture Medium That Enhances Propionibacterium freudenreichii Stress Tolerance.

UNLABELLED: Propionibacterium freudenreichii is used as a cheese-ripening starter and as a probiotic. Its reported physiological effects at the gut level, including modulation of bifidobacteria, colon epithelial cell proliferation and apoptosis, and intestinal inflammation, rely on active metabolism in situ Survival and activity are thus key factors determining its efficacy, creating stress adaptation and tolerance bottlenecks for probiotic applications. Growth media and growth conditions determine tolerance acquisition. We investigated the possibility of using sweet whey, a dairy by-product, to sustain P. freudenreichii growth. It was used at different concentrations (dry matter) as a culture medium. Using hyperconcentrated sweet whey led to enhanced multistress tolerance acquisition, overexpression of key stress proteins, and accumulation of intracellular storage molecules and compatible solutes, as well as enhanced survival upon spray drying. A simplified process from growth to spray drying of propionibacteria was developed using sweet whey as a 2-in-1 medium to both culture P. freudenreichii and protect it from heat and osmotic injury without harvesting and washing steps. As spray drying is far cheaper and more energy efficient than freeze-drying, this work opens new perspectives for the sustainable development of new starter and probiotic preparations with enhanced robustness. IMPORTANCE: In this study, we demonstrate that sweet whey, a dairy industry by-product, not only allows the growth of probiotic dairy propionibacteria, but also triggers a multitolerance response through osmoadaptation and general stress response. We also show that propionibacteria accumulate compatible solutes under these culture conditions, which might account for the limited loss of viability after spray drying. This work opens new perspectives for more energy-efficient production of dairy starters and probiotics.

Shape, shell, and vacuole formation during the drying of a single concentrated whey protein droplet.

The drying of milk concentrate droplets usually leads to specific particle morphology influencing their properties and their functionality. Understanding how the final shape of the particle is formed therefore represents a key issue for industrial applications. In this study, a new approach to the investigation of droplet-particle conversion is proposed. A single droplet of concentrated globular proteins extracted from milk was deposited onto a hydrophobic substrate and placed in a dry environment. Complementary methods (high-speed camera, confocal microscopy, and microbalance) were used to record the drying behavior of the concentrated protein droplets. Our results showed that whatever the initial concentration, particle formation included three dynamic stages clearly defined by the loss of mass and the evolution of the internal and external shapes of the droplet. A new and reproducible particle shape was related in this study. It was observed after drying a smooth, hemispherical cap-shaped particle, including a uniform protein shell and the nucleation of an internal vacuole. The particle morphology was strongly influenced by the drying environment, the contact angle, and the initial protein concentration, all of which governed the duration of the droplet shrinkage, the degree of buckling, and the shell thickness. These results are discussed in terms of specific protein behaviors in forming a predictable and a characteristic particle shape. The way the shell is formed may be the starting point in shaping particle distortion and thus represents a potential means of tuning the particle morphology.

Physicochemical, techno-functional, and fat melting properties of spray-dried camel and bovine milk powders.

In this study, three skimmed and one whole-fat spray-dried camel milk powders were produced and their characteristics were compared to those of bovine milk powders. The physicochemical analysis of the produced powders indicated that camel milk powders (whether skimmed or not) presented higher ash and whey protein contents as compared to those of bovine milk powders. Our results indicated that the investigated camel and bovine milk powders exhibited a high solubility index (>99%) with poor dispersibility and wettability indexes due to their small particles size (d50 ≤ 12 µm) and their narrow size distribution (span ≤ 2). In addition, although camel and bovine milk powders presented the same total fat content, lower free fat content was measured for camel milk powders. Besides, the whey protein nitrogen index and the SDS-PAGE electrophoresis underlined that camel and bovine milk proteins remained intact after drying with low denaturation extent. It is worth noticed that camel milk proteins were less denaturized due to the absence of the heat-sensitive ß-lactoglobulin in camel milk. Moreover, the low denaturation extent participated in the enhancing of the foaming capacity and stability of camel and bovine milk powders. Finally, the calorimetric analysis showed that higher fat melting temperatures were recorded in whole-fat camel milk powder and in their anhydrous form as compared to those of bovine milk. PRACTICAL APPLICATION: Camel milk powder is an emerging non-bovine dairy product. Understanding its rehydration ability and evaluating the impact of spray drying on its protein quality are promising approaches to obtain high-quality camel milk powder with high reconstitution ability. Findings of this study indicated that spray drying is a suitable technique to produce highly soluble camel milk powders with low denaturation extent. These results will benefit the research and development department of food industry (especially those producing camel milk powder) as well as the direct consumers.

Effect of pH on the physicochemical characteristics and the surface chemical composition of camel and bovine whey protein's powders.

This study investigated the effect of pH on the denaturation extent, the surface chemical composition, the water sorption isotherm and the glass transition temperature of camel and bovine whey protein's powders. The LC-MS analysis indicated that the ß-Lactoglobulin was the most denatured protein in bovine whey powders regardless the pH value, while this protein was totally absent in camel whey. The α-Lactalbumin was relatively heat stable after drying and predominated the powder surface (X-ray photoelectron spectroscopy results) in both camel and bovine whey powders regardless the pH (neutral (6.7) or acidic (4.3 and 4.6)). Analysis of the water sorption isotherms indicated that decreasing the pH induced the increase of the water activity of lactose crystallization for camel and bovine whey powders. Finally, decreasing the pH led to the decrease of the glass transition temperature of camel and bovine whey powder (at 0.13, 0.23, and 0.33 of water activity).

Relationships between dairy powder surface composition and wetting properties during storage: importance of residual lipids.

The relationships between powder surface composition and powder rehydration properties under variable conditions of storage are investigated in this paper. A rheological approach was used to evaluate the modifications induced by storage on the rehydration properties of native phosphocaseinate powder. Concurrently, the powder surface composition (i.e., lactose, proteins, and lipids) was evaluated by X-ray photoelectron spectroscopy (XPS). A strong correlation was found between the powder wetting time lengthening and the migration of lipids on the powder surface during storage. XPS studies indicated also an over-representation of lipids on the powder surface (6%) in comparison with total lipids (0.4%) even on fresh powder before storage. Detailed investigation of powder lipids revealed the presence of high levels of polar lipids (66% compared with <1% in milk lipids). Their amphiphilic nature and their melting points could explain the extensive enrichment of lipids observed at the powder surface during processing and storage.

Technological aspects of lactose-hydrolyzed milk powder.

Few reports describe the effect of lactose hydrolysis on the properties of milk powder during production and storage. Hence, the aim of this study was to evaluate the effects of five different levels of enzymatic lactose hydrolysis during the production and storage of milk powder. As the lactose hydrolysis rate increased, adhesion to the drying chamber also increased, due to higher levels of particle agglomeration. Additionally, more brown powder was obtained when the lactose hydrolysis rate was increased, which in turn negatively affected rehydration ability. Using Raman spectroscopy, crystallization of the lactose residues in various samples was assessed over 6weeks of accelerated aging at a room temperature environment with 75.5% of air moisture. Products with 25% or greater lactose hydrolysis showed no signs of crystallization, in contrast to the non-hydrolyzed sample.

The influence of cheese composition and microstructure on the diffusion of macromolecules: A study using Fluorescence Recovery After Photobleaching (FRAP).

In cheese technology, the diffusion phenomena are crucial during ripening. The technique of Fluorescence Recovery After Photobleaching was applied for the first time on real cheese, in order to investigate the relationships between molecular diffusion and the cheese composition and/or its microstructure. Measured effective diffusion coefficients in soft and hard cheese of a group of dextrans (10-500 kDa) were found to be in the same order of magnitude with values observed when using a comparable non-fat model cheese (∼ 0.1-20 µm(2) s(-1)). Diffusion of the dextrans was mainly dependent on the fraction of "free" aqueous phase present in the cheese, closely which is linked to cheese-making technology and ripening stage. Diffusion coefficients were modeled by a power law relationship as a function of dextran molecular weight, which allowed some study of the cheese microstructure. A tighter protein network will require some deformation of those flexible macromolecules with a higher molecular weight (>250 kDa), in order to diffuse through the pores of such cheese structures.