Can encapsulation lengthen the shelf-life of probiotic bacteria in dry products?

This study evaluated the use of microencapsulation to maintain probiotic Lactobacillus rhamnosus GG (LGG) viability during exposure to detrimentally high levels of water activity. Freeze-dried LGG was mixed into a whey protein solution and sprayed onto a core particle. A second batch was prepared by applying an extra layer of palm oil onto the LGG encapsulates. Viability of the LGG preparations was measured over time in an accelerated shelf-life study at 37 degrees C which included exposure to different water activities (a(w)=0.15 or 0.7), ambient oxygen, and incorporation of the probiotic in infant formula powder. Results showed that a high water activity (0.7) was detrimental to the survival of unencapsulated LGG, yielding more than a 10 log(10) loss in cell viability within 2 weeks. The presence of oxygen and infant formula conferred no additional effects. Encapsulation of LGG did not improve the survival of this strain under the conditions tested here. Although encapsulation is often mentioned as a quick-fix for preserving probiotic viability in dry and semi-dry products, more studies are needed which validate the broader potential of this approach at different shelf-life expectancies, water activities, and product matrices.

In vitro analysis of protection of the enzyme bile salt hydrolase against enteric conditions by whey protein-gum arabic microencapsulation.

The interest in efficient intestinal delivery of health-promoting substances is increasing. However, the delivery of vulnerable substances such as enzymes requires specific attention. The transit through the stomach, where the pH is very low, can be detrimental to the enzymatic activity of the protein to be delivered. Here, we describe the microencapsulation of the model enzyme bile salt hydrolase (Bsh) using whey protein-gum arabic microencapsulates for food-grade and targeted enzyme delivery in the proximal region of the small intestine. Furthermore, the efficacy of enteric coating microencapsulates for site-specific enzyme delivery was compared in vitro with living Lactobacillus plantarum WCFS1 bacteria that endogenously produce the Bsh enzyme. Microencapsulates allowed highly effective protection of the enzyme under gastric conditions. Moreover, Bsh release under intestinal conditions appeared to be very efficient, although in the presence of pancreatin, the Bsh activity decreased in time due to proteolytic degradation. In comparison, L. plantarum appeared to be capable to withstand gastric conditions as well as pancreatin challenge. Delivery using encapsulates and live bacteria each have different (dis)advantages that are discussed. In conclusion, live bacteria and food-grade microencapsulates provide alternatives for dedicated enteric delivery of specific enzymes, and the choice of enzyme to be delivered may determine which mode of delivery is most suitable.

Composition and structure of whey protein/gum arabic coacervates.

Complex coacervation in whey protein/gum arabic (WP/GA) mixtures was studied as a function of three main key parameters: pH, initial protein to polysaccharide mixing ratio (Pr:Ps)(ini), and ionic strength. Previous studies had already revealed under which conditions a coacervate phase was obtained. This study is aimed at understanding how these parameters influence the phase separation kinetics, the coacervate composition, and the internal coacervate structure. At a defined (Pr:Ps)(ini), an optimum pH of complex coacervation was found (pH(opt)), at which the strength of electrostatic interaction was maximum. For (Pr:Ps)(ini) = 2:1, the phase separation occurred the fastest and the final coacervate volume was the largest at pH(opt) = 4.0. The composition of the coacervate phase was determined after 48 h of phase separation and revealed that, at pH(opt), the coacervate phase was the most concentrated. Varying the (Pr:Ps)(ini) shifted the pH(opt) to higher values when (Pr:Ps)(ini) was increased and to lower values when (Pr:Ps)(ini) was decreased. This phenomenon was due to the level of charge compensation of the WP/GA complexes. Finally, the structure of the coacervate phase was studied with small-angle X-ray scattering (SAXS). SAXS data confirmed that at pH(opt) the coacervate phase was dense and structured. Model calculations revealed that the structure factor of WP induced a peak at Q = 0.7 nm(-1), illustrating that the coacervate phase was more structured, inducing the stronger correlation length of WP molecules. When the pH was changed to more acidic values, the correlation peak faded away, due to a more open structure of the coacervate. A shoulder in the scattering pattern of the coacervates was visible at small Q. This peak was attributed to the presence of residual charges on the GA. The peak intensity was reduced when the strength of interaction was increased, highlighting a greater charge compensation of the polyelectrolyte. Finally, increasing the ionic strength led to a less concentrated, a more heterogeneous, and a less structured coacervate phase, induced by the screening of the electrostatic interactions.

Microencapsulation of oils using whey protein/gum Arabic coacervates.

Microencapsulating sunflower oil, lemon and orange oil flavour was investigated using complex coacervation of whey protein/gum arabic (WP/GA). At pH 3.0-4.5, WP and GA formed electrostatic complexes that could be successfully used for microencapsulation purposes. The formation of a smooth biopolymer shell around the oil droplets was achieved at a specific pH (close to 4.0) and the payload of oil (i.e. amount of oil in the capsule) was higher than 80%. Small droplets were easier to encapsulate within a coacervate matrix than large ones, which were present in a typical shell/core structure. The stability of the emulsion made of oil droplets covered with coacervates was strongly pH-dependent. At pH 4.0, the creaming rate of the emulsion was much higher than at other pH values. This phenomenon was investigated by carrying out zeta potential measurements on the mixtures. It seemed that, at this specific pH, the zeta potential was close to zero, highlighting the presence of neutral coacervate at the oil/water interface. The influence of pH on the capsule formation was in accordance with previous results on coacervation of whey proteins and gum arabic, i.e. WP/GA coacervates were formed in the same pH window with and without oil and the pH where the encapsulation seemed to be optimum corresponded to the pH at which the coacervate was the most viscous. Finally, to illustrate the applicability of these new coacervates, the release of flavoured capsules incorporated within Gouda cheese showed that large capsules gave stronger release and the covalently cross-linked capsules showed the lowest release, probably because of a tough dense biopolymer wall which was difficult to break by chewing.

Complex coacervation of whey proteins and gum arabic.

Mixtures of gum arabic and whey protein (whey protein isolate, WP) form an electrostatic complex in a specific pH range. Three phase boundaries (pH(c), pHphi(1), pHphi(2)) have been determined using an original titration method, newly applied to complex coacervation. It consists of monitoring the turbidity and light scattering intensity under slow acidification in situ with glucono-delta-lactone. Furthermore, the particle size could also be measured in parallel by dynamic light scattering. When the pH is lowered, whey proteins and gum arabic first form soluble complexes. This boundary is designated as pH(c). When the interaction is stronger (at lower pH), phase separation takes place (at pHphi(1)). Finally, at pHphi(2) complexation was suppressed by the charge reduction of the gum arabic. The major constituent of the whey protein preparation used was beta-lactoglobulin (beta-lg), and it was shown that beta-lg was indeed the main complex-forming protein. Moreover, an increase of the ionic strength shifted the pH boundaries to lower pH values, which was summarized in a state diagram. The experimental pH(c) values were compared to a newly developed theory for polyelectrolyte adsorption on heterogeneous surfaces. Finally, the influence of the total biopolymer concentration (0-20% w/w) was represented in a phase diagram. For concentrations below 12%, the results are consistent with the theory on complex coacervation developed by Overbeek and Voorn. However, for concentrations above 12%, phase diagrams surprisingly revealed a "metastable" region delimited by a percolation line. Overall, a strong similarity is seen between the behavior of this system and a colloidal gas-liquid phase separation.