Co-encapsulation of Omega-3 fatty acids and probiotic bacteria through complex coacervation

The research described in this thesis investigated the microencapsulation of omega-3 oil and probiotic bacteria together in a protein-polysaccharide complex coacervate matrix. The synergistic or competitive interactions between the probiotic bacteria and omega-3 fatty acids when packaged in a single microcapsule was determined including how best to utilise such interaction to achieve improved oxidative stability of omega-3 fatty acid and better survival of the probiotic bacteria. Encapsulation and co-encapsulation of tuna oil (O) and Lactobacillus casei 431 (P) as models of omega-3 and probiotic bacteria, respectively, were carried out and the works is described in this thesis in five distinct sections. (1) The optimisation of the complex coacervation process between whey protein isolate (WPI) and gum Arabic (GA). (2) Microencapsulation of tuna oil (O) in WPI-GA complex coacervates followed by spray and freeze drying to produce microcapsules (WPI-O-GA). (3) Microencapsulation of probiotic bacteria L. casei 431 (P) in WPI-GA complex coacervates followed by spray and freeze drying to produce microcapsules (WPI-P-GA). (4) Co-encapsulation of omega-3 oil and L. casei 431 together in WPI-GA coacervate matrix followed by spray and freeze drying to produce co-microcapsules (WPI-P-O-GA). (5) In-vitro digestion evaluation of co-microcapsules and microcapsules to indicate bioavailability. The viability of L. casei was significantly higher in WPI-P-O-GA co-microcapsules than in WPI-P-GA microcapsules in both spray and freeze dried microcapsules. The oxidative stability of tuna oil was significantly higher in spray dried co-capsules. Also, co-microencapsulation increased the survivability of L. casei during simulated digestion. There was no significant influence observed on the release properties of omega-3 oil due to co-microencapsulation. However, the total omega-3 fatty acids in the released oil during in-vitro digestion were found to be higher, when co-microencapsulated. Hence, co-microencapsulation was shown to protect the L. casei and deliver both viable cells and omega-3 oil to human intestine without any significant adverse effect on their functionality and properties.Doctor of Philosoph

Physiological protection of probiotic microcapsules by coatings

Nowadays, food and nutrition have a greater impact in people's concerns, with the awareness that nutrition have a direct impact in health and wellbeing. Probiotics have an important role in this topic and consumers are starting to really understand their potential in health, leading to an increasing interest of the companies to their commercial use in foods. However, there are several limitations to the use of probiotics in foods and beverages, being one of them their efficiency (directly associated to their survival rate) upon ingestion. This work is focused in microencapsulation techniques that have been used to increase probiotics efficiency. More specifically, this work reviews the most recent and relevant research about the production and coating techniques of probiotic-loaded microcapsules, providing an insight in the effect of these coatings in probiotics survival during the gastrointestinal phase. This review shows that coatings with the better performances in probiotics protection, against the harsh conditions of digestion, are chitosan, alginate, poly-L-lysine and whey protein. Chitosan presented an interesting performance in probiotics protection being able to maintain the initial concentration of viable probiotics during a digestive test. The analyses of different works also showed that the utilization of several coatings does not guarantee a better protection in comparison with monocoated microcapsules.The author Philippe E. Ramos is recipient of fellowships from the Fundação para a Ciência e Tecnologia, POPH-QREN and FSE (FCT, Portugal) through the grant SFRH/BD/80800/2012. This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684). RECI Project (Until December of 2017): This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the Project RECI/BBBEBI/0179/2012 (FCOMP-01-0124-FEDER-027462).info:eu-repo/semantics/publishedVersio

INFOGEST static in vitro simulation of gastrointestinal food digestion

peer-reviewedSupplementary information is available at http://dx.doi.org/10.1038/s41596-018-0119-1 or https://www.nature.com/articles/s41596-018-0119-1#Sec45.Developing a mechanistic understanding of the impact of food structure and composition on human health has increasingly involved simulating digestion in the upper gastrointestinal tract. These simulations have used a wide range of different conditions that often have very little physiological relevance, and this impedes the meaningful comparison of results. The standardized protocol presented here is based on an international consensus developed by the COST INFOGEST network. The method is designed to be used with standard laboratory equipment and requires limited experience to encourage a wide range of researchers to adopt it. It is a static digestion method that uses constant ratios of meal to digestive fluids and a constant pH for each step of digestion. This makes the method simple to use but not suitable for simulating digestion kinetics. Using this method, food samples are subjected to sequential oral, gastric and intestinal digestion while parameters such as electrolytes, enzymes, bile, dilution, pH and time of digestion are based on available physiological data. This amended and improved digestion method (INFOGEST 2.0) avoids challenges associated with the original method, such as the inclusion of the oral phase and the use of gastric lipase. The method can be used to assess the endpoints resulting from digestion of foods by analyzing the digestion products (e.g., peptides/amino acids, fatty acids, simple sugars) and evaluating the release of micronutrients from the food matrix. The whole protocol can be completed in ~7 d, including ~5 d required for the determination of enzyme activities.COST action FA1005 INFOGEST (http://www.cost-infogest.eu/ ) is acknowledged for providing funding for travel, meetings and conferences (2011-2015). The French National Institute for Agricultural Research (INRA, www.inra.fr) is acknowledged for their continuous support of the INFOGEST network by organising and co-funding the International Conference on Food Digestion and workgroup meeting

Recent advances in the microencapsulation of omega-3 oil and probiotic bacteria through complex coacervation: a review

Background Functional foods are a fastest growing sector of the food industry. The development of functional foods comprising omega-3 fatty acids and probiotic bacteria, through complex coacervation process is an emerging area of research and product development. Scope and approach We reviewed relevant literature concerning the use of complex coacervation in microencapsulation, focusing primarily on the inclusion of probiotic bacteria and omega-3 oils into a single delivery format. This review covers advantages and disadvantages of the complex coacervation process to microencapsulate bioactive ingredients, viability of probiotic bacteria and oxidative stability of omega-3 oil during the complex coacervation process, the bioaccessibility of omega-3 oil and probiotic bacteria during simulated gastrointestinal conditions and in-vivo testings. Key findings and conclusions The review describes the advantages of co-encapsulation using complex coacervation followed by spray drying. It also describes the technological hurdles that need to be resolved for further development of industrial applications of co-encapsulation of probiotic bacteria and omega-3 lipids. The co-encapsulation concept has been widely used in pharmaceutical delivery systems, but is a relatively new concept in food ingredient stabilisation and delivery. There is a commercial need of co-encapsulation of multiple bioactive ingredients within a single microcapsules, due to decreased cost and enhanced product quality. Complex coacervation has been shown to be a useful method for the co-encapsulation of multiple unstable bioactive ingredients. Although in-vitro evaluation deliver useful bioavailability information, additional in-vivo and clinical trials are needed to determine the efficacy of bioactive release, particularly for microcapsules containing multiple bioactive ingredients

Survival and fermentation activity of probiotic bacteria and oxidative stability of omega-3 oil in co-microcapsules during storage

Tuna oil (O) and probiotic bacteria Lactobacillus casei (P) were co-microencapsulated in whey protein isolate (WPI)-gum Arabic (GA) complex coacervate. The co-microcapsules (WPI-P-O-GA), L. casei microcapsules (WPI-P-GA) and tuna oil microcapsules (WPI-O-GA) were converted into powder using spray and freeze drying. The interaction between probiotic bacteria and omega-3 oil in co-microcapsules, particularly in terms of oxidative stability of omega-3 oil and vitality/viability of probiotic bacteria and any synergistic outcome, was studied. The effect of storage temperature (5 and 25 °C) and time (90 days) on the survival and fermentation activity of L. casei and oxidative stability of tuna oil in the microcapsules/co-microcapsules was determined. A synergism between oxidative stability of omega-3 oil and vitality of probiotic bacteria was observed, when they were co-microencapsulated and spray dried. These co-microcapsules will likely have utility in functional food formulations due to simple and cost effective stabilisation and delivery of two important functional ingredients

Complex coacervation with whey protein isolate and gum arabic for the microencapsulation of omega-3 rich tuna oil

Tuna oil rich in omega-3 fatty acids was microencapsulated in whey protein isolate (WPI)-gum arabic (GA) complex coacervates, and subsequently dried using spray and freeze drying to produce solid microcapsules. The oxidative stability, oil microencapsulation efficiency, surface oil and morphology of these solid microcapsules were determined. The complex coacervation process between WPI and GA was optimised in terms of pH, and WPI-to-GA ratio, using zeta potential, turbidity, and morphology of the microcapsules. The optimum pH and WPI-to-GA ratio for complex coacervation was found to be 3.75 and 3:1, respectively. The spray dried solid microcapsules had better stability against oxidation, higher oil microencapsulation efficiency and lower surface oil content compared to the freeze dried microcapsules. The surface of the spray dried microcapsules did not show microscopic pores while the surface of the freeze dried microcapsules was more porous. This study suggests that solid microcapsules of omega-3 rich oils can be produced using WPI-GA complex coacervates followed by spray drying and these microcapsules can be quite stable against oxidation. These microcapsules can have many potential applications in the functional food and nutraceuticals industry

In-vitro digestion of probiotic bacteria and omega-3 oil co-microencapsulated in whey protein isolate-gum Arabic complex coacervates

Solid co-microcapsules of omega-3 rich tuna oil and probiotic bacteria L. casei were produced using whey protein isolate-gum Arabic complex coacervate as wall material. The in-vitro digestibility of the co-microcapsules and microcapsules was studied in terms of survival of L. casei and release of oil in sequential exposure to simulated salivary, gastric and intestinal fluids. Co-microencapsulation significantly increased the survival and surface hydrophobicity and the ability of L. casei to adhere to the intestinal wall. No significant difference in the assimilative reduction of cholesterol was observed between the microencapsulated and co-microencapsulated L. casei. The pattern of release of oil from the microcapsules and co-microcapsules was similar. However, the content of total chemically intact omega-3 fatty acids was higher in the oil released from co-microcapsules than the oil released from microcapsules. The co-microencapsulation can deliver bacterial cells and omega-3 oil to human intestinal system with less impact on functional properties. © 2017 Elsevier Lt

Co-encapsulation and characterisation of omega-3 fatty acids and probiotic bacteria in whey protein isolate-gum Arabic complex coacervates

Omega-3 fatty acids and probiotic bacteria were co-encapsulated in a single whey protein isolate (WPI)-gum Arabic (GA) complex coacervate microcapsule. Tuna oil (O) and Lactobacillus casei 431 (P) were used as models of omega-3 and probiotic bacteria, respectively. The co-microcapsules (WPI-P-O-GA) and L.casei containing microcapsules (WPI-P-GA) were converted into powder by using spray and freeze drying. The viability of L.casei was significantly higher in WPI-P-O-GA co-microcapsules than in WPI-P-GA. The oxidative stability of tuna oil was significantly higher in spray dried co-capsules than in freeze dried ones