Survival of probiotics encapsulated in chitosan-coated alginate beads in yoghurt from UHT- and conventionally treated milk during storage

Survival of the microencapsulated probiotics, Lactobacillus acidophilus 547, Bifidobacterium bifidum ATCC 1994, and Lactobacillus casei 01, in stirred yoghurt from UHT- and conventionally treated milk during low temperature storage was investigated. The probiotic cells both as free cells and microencapsulated cells (in alginate beads coated with chitosan) were added into 20 g/100 g total solids stirred yoghurt from UHT-treated milk and 16 g/100 g total solids yoghurt from conventionally treated milk after 3.5 h of fermentation. The products were kept at 4 °C for 4 weeks. The survival of encapsulated probiotic bacteria was higher than free cells by approximately 1 log cycle. The number of probiotic bacteria was maintained above the recommended therapeutic minimum (10⁷ cfu g⁻¹) throughout the storage except for B. bifidum. The viabilities of probiotic bacteria in yoghurts from both UHT- and conventionally treated milks were not significantly (P>0.05) different.     

Alginate beads and apple pieces as carriers for Saccharomyces cerevisiae var. boulardii,as representative of yeast functional starter cultures

This study focused on two different carriers for Saccharomyces cerevisiae var. boulardii for its targeted release as a starter or as a probiotic: the microencapsulation into alginate beads and the immobilisation on apple pieces. Beads were studied in relation to encapsulation yield (EY), viability of cells throughout the storage, kinetic of cell release, survival under conditions that mimic the transit into the gut and in vivo application (fermentation of grape juice). Concerning the microencapsulation, EY was very high (ca. 90%), cells survived for a long period (upon to 90 days), and the beads assured cell survival and their release under conditions that mimic the gut; moreover, the capsules could be used up to ten times to start a model fermentation of grape juice. Apple pieces were a good system for the immobilisation of S. boulardii; they could be proposed successfully as a reusable carrier for a starter culture, as they assured the start of the fermentation of grape juice for at least seven times.     

Effect of addition of inulin and galactooligosaccharide on the survival of microencapsulated probiotics in alginate beads coated with chitosan in simulated digestive system,yogurt and fruit juice

Galactooligosaccharides (GOS) and inulin were added during microencapsulation of Lactobacillus acidophilus 5 and Lactobacillus casei 01 in alginate beads coated with chitosan at the concentrations of 0, 0.5, 1.0 and 1.5%. Addition of prebiotics significantly (p < 0.05) increased the bead size by approximately 3.8%. The presence of GOS (0.3%) in the microencapsulation provided the best protection with only 3.1 and 2.9 logs reduction for L. acidophilus 5 and L. casei 01, respectively, after incubation in simulated gastric juice (pH 1.55), followed by simulate intestinal juice containing 0.6% bile salt. The viabilities of microencapsulated probiotics containing 1.5% GOS in commercial yogurt and orange juice were also performed at refrigerated storage for 4 weeks. In yogurt, the numbers of cells with GOS were higher than those of without GOS by approximately 1.1 and 0.4 logs for L. acidophilus 5 and L. casei 01, respectively. In orange juice, the numbers of cells with GOS were higher than those of without GOS by approximately 0.5 and 0.4 logs for L. acidophilus 5 and L. casei 01, respectively. The numbers of probiotic bacteria were maintained above the recommended therapeutic minimum (10⁷ cfu g⁻¹ or mL⁻¹ of product) throughout the storage in both products.     

山羊乳-FOS/GOS库德毕赤酵母DS8-1微胶囊的制备及体外评价

前期从新疆传统乳制品酸驼乳中筛选得到一株具有潜在益生特性的库德毕赤酵母DS8-1（Pichia kudriavzevii DS8-1）,为增加菌株对体外环境的抗性,以海藻酸钠为壁材使用锐孔法制备酵母菌微胶囊。将干燥的藻酸盐（SA）微胶囊外包山羊乳或在微胶囊壁材中添加低聚果糖（fructooligosaccharides,FOS）/低聚半乳糖（galactooligosaccharides,GOS）分别制备3 种酵母菌微胶囊：SAM微胶囊、SAMF微胶囊和SAMG微胶囊。对微胶囊进行傅里叶红外光谱和扫描电子显微镜表征、模拟胃液的耐受性、消化液的释放特性和贮藏稳定性实验,研究冻干微胶囊对菌株活性的影响。结果表明：使用羊乳包衣的3 种冻干微胶囊（SAM微胶囊、SAMF微胶囊和SAMG微胶囊）的菌株活力为7.16～7.67（lg（CFU/g））。与SA微胶囊相比,SAMF微胶囊在连续的模拟消化液处理后能够延缓酵母菌的释放,提高菌株存活率,以及对α-淀粉酶和α-葡萄糖苷酶具有较高的抑制活性。SAMG微胶囊在－20、4、25 ℃贮藏30 d后菌株活力在7（lg（CFU/g））以上。结论：在海藻酸钠中添加FOS/GOS且使用羊乳包衣可用于包埋库德毕赤酵母,制备的酵母菌微胶囊具有提高菌株活性的作用,在功能性食品输送体系中具有一定的应用前景。     

Stability of Lactobacillus reuteri in Different Types of Microcapsules

This study was designed to find the most suitable method and wall material for microencapsulation of the probiotic bacterium Lactobacillus reuteri to maintain cell viability during gastric challenge. Five L. reuteri strains were individually encapsulated using alginate, alginate plus starch, K‐carrageenan with locust bean gum, or xanthan with gellan by extrusion or phase separation (emulsion). The morphology of the microcapsules was studied using phase contrast and cryo‐scanning electron microscopy (cryo‐SEM). The resistance of these microcapsules and the viability of contained L. reuteri to simulated gastric juice were studied. The shape and size of the microcapsules produced varied with the preparation method and type of wall material. Extruded microcapsules were larger and more uniformly shaped. Survival of microencapsulated L. reuteri was significantly better than that of planktonic cells and varied with the strain, method of microencapsulation, and wall material used. In general, microencapsulation using alginate and alginate with starch by both extrusion and phase separation were found to provide bacteria significantly greater protection (P < 0.05) against simulated gastric juice.     

A new approach to eliminate stress for two probiotics with chemicals in vitro

The effect of some chemicals to eliminate stress for two probiotics cultivation was studied. Lactobacillus acidophilus was incubated with sodium citrate or calcium carbonate at 37 °C for 19 h, or Bifidobacterium bifidum was incubated with sodium d-isoascorbate or sodium ascorbate at 37 °C for 24 h in an anaerobic chamber, aiming to eliminate acidic or oxidative stress produced during culture. Viable count of L. acidophilus or B. bifidum in culture was counted with standard plate counting technique. The result showed that the viable count of L. acidophilus or B. bifidum in culture had an increase about twofold compared with that of control. The mechanism involved was discussed preliminary. Chemical destressing effect of sodium citrate and calcium carbonate on L. acidophilus, or chemical destressing effect of sodium d-isoascorbate and sodium ascorbate on B. bifidum, might provide a potential approach for probiotic production.     

海藻酸钠二次包衣对益生菌微胶囊包埋效果的影响研究

为了考察海藻酸钠二次包衣对两歧双歧杆菌F-35包埋效果的影响,本研究分别制备得到了海藻酸钠微胶囊（AL-M）、蛋白质微胶囊（WP-M）和海藻酸钠二次包衣的微胶囊（AL-CM）,并从三个方面（壁材降解速度、益生菌释放速度和益生菌存活情况）对这三种微胶囊在连续模拟胃肠液环境下的降解进行了比较。结果表明,海藻酸钠二次包衣能够显著降低胃蛋白酶对乳清蛋白的降解作用。相比于AL-M和WP-M,AL-CM会延缓两歧双歧杆菌F-35在模拟肠液中的释放速率,并且在连续的模拟胃肠道中经过二次包以后的微胶囊中对两歧双歧杆菌F-35保护效果最好,两歧双歧杆菌F-35最终的存活率高达51.0%。      

An Improved Method of Microencapsulation of Probiotic Bacteria for Their Stability in Acidic and Bile Conditions during Storage

ABSTRACT: The purpose of this study was to develop a method for applying an extra coating of palm oil and poly‐L‐lysine (POPL) to alginate (ALG) microcapsules to enhance the survival of probiotic bacteria. Eight strains of probiotic bacteria including Lactobacillus rhamnosus, Bifidobacterium longum, L. salivarius, L. plantarum, L. acidophilus, L. paracasei, B. lactis type Bl‐O4, and B. lactis type Bi‐07 were encapsulated using alginate alone or alginate with POPL. Electron microscopy was used to measure the size of the microcapsules and to determine their surface texture. To assess if the addition of POPL improved the viability of probiotic bacteria in acidic conditions, both ALG and POPL microcapsules were inoculated into pH 2.0 MRS broths and their viability was assessed over a 2‐h incubation period. Two bile salts including oxgall bile salt and taurocholic acid were used to test the bile tolerance of probiotic bacteria entrapped in ALG and POPL microcapsules. To assess the porosity and the ability of the microcapsule to hold small molecules in an aqueous environment a water‐soluble fluorescent dye, 6‐carboxyflourescin (6 FAM), was encapsulated and its release was monitored using a UV spectrophotometer. The results indicated that coating the microcapsules with POPL increased the overall size of the capsules by an average of 3 μm ± 0.67. However, microcapsules with added POPL had a much smoother surface texture when examined under an electron microscope. The results also indicated that the addition of POPL to microcapsules improved the average viability of probiotic bacteria by > 1 log CFU/mL when compared to ALG microcapsules at 2 h of exposure to acidic conditions. However, similar plate counts were observed between ALG and POPL microcapsules when exposed to bile salts. This suggests that an extra coating of POPL could be readily broken down by bile salts that are commonly found in the lower gastrointestinal tract (GIT). Upon testing the porosity of the microcapsules, findings suggest that POPL microcapsules were less porous and hold 52.2% more fluorescent dye over a 6‐wk storage period.     

The influence of multi stage alginate coating on survivability of potential probiotic bacteria in simulated gastric and intestinal juice

The probiotics, Lactobacillus acidophilus PTCC1643 and Lactobacillus rhamnosus PTCC1637, were encapsulated into uncoated calcium alginate beads and the same beads were coated with one or two layers of sodium alginate with the objective of enhancing survival during exposure to the adverse conditions of the gastro-intestinal tract. The survivability of the strains, was expressed as the destructive value (decimal reduction time). Particle size distribution was measured using laser diffraction technique. The thickness of the alginate beads increased with the addition of coating layers. No differences were detectable in the bead appearance by scanning electron microscopy (SEM). The alginate coat prevented acid-induced reduction of the strains in simulated gastric juice (pH 1.5, 2 h), resulting in significantly (P < 0.05) higher numbers of survivors. After incubation in simulated gastric (60 min) and intestinal juices (pH 7.25, 2 h), number of surviving cells were 6.5 log cfu mL^?1 for L. acidophilus and 7.6 log cfu mL^?1 for L. rhamnosus by double layer coated alginate microspheres, respectively, while 2.3 and 2.0 log cfu mL^?1 were obtained for free cells, respectively.     

Shelf life and quality of probiotic yogurt produced with Lactobacillus acidophilus and Gobdin

This study evaluated the effect of dry white mulberry and walnut paste (Gobdin, a traditional Turkish food) in probiotic yogurt on the survival of Lactobacillus acidophilus and yogurt properties. Six different yogurts were produced with 0%, 5% and 10% Gobdin using Lactobacillus bulgaricus + Streptococcus thermophilus and with 0%, 5% and 10% Gobdin using L. bulgaricus + S. thermophilus + L. acidophilus. The physical, chemical, microbiological and sensorial properties of the yogurts were evaluated based on storage at 4 ± 1 °C. Probiotic shelf life and the most suitable combinations were determined. The highest L. acidophilus count (8.65 log cfu g^?1) was found in the 5% Gobdin‐supplemented yogurt on the 7th day of storage, while the lowest count (8.11 log cfu g^?1) was found in the probiotic control yogurt on the 21st day. Although the L. acidophilus counts in the probiotic yogurts declined during storage, all values found throughout the 21‐day storage period were >8 log cfu g^?1. This is above the level necessary to provide the desired therapeutic effect in probiotic products (10⁶–10⁷ cfu g^?1). The highest overall acceptability score was obtained on the first day from the yogurt with 5% Gobdin. However, all yogurt samples had general acceptability scores between 7 and 8 points from a 9‐point maximum. Thus, this study determined that a new functional yogurt can be produced using L. acidophilus with 5% Gobdin.     

Effect of different levels of fat and inulin on the microbial growth and metabolites in probiotic yogurt containing nonviable bacteria

Effects of different levels of fat and inulin on bacterial cell counts, degree of proteolysis and concentrations of organic acids in the yogurt containing inactivated cells of probiotic strains Bifidobacterium animalis and Lactobacillus acidophilus were investigated. Results showed that both L. acidophilus and B. animalis grew well in the yogurt samples reaching cell counts higher than 10⁶ CFU mL^?1 at the final pH of 4.5. Inulin at the concentration of 1% had no significant effects on the production of organic acids and cell counts of L. acidophilus, but promoted the growth of B. animalis with a reduction in the degree of proteolysis. Generally, different fat levels showed significant effects on the production of organic acids and nonsignificant effects on the cell counts of probiotic bacteria and degree of proteolysis. In case of lactic acid, the ratio of L‐ (+)to D‐ (?) isomer ranged from 50/50 to 80/20 in yogurt samples.     

Survival of probiotic microflora in Argentinian yoghurts during refrigerated storage

The survival of Bifidobacterium bifidum BBI and Lactobacillus acidophilus LAI in reduced-fat (liquid) and full-fat (set) yoghurts produced with two commercial lactic starter cultures (SID and SISD) was investigated. The viability of the probiotic bacteria was also assayed in milk acidified with lactic acid at different pH values. Samples were stored at 5°C for up to 4 weeks. There was a great variability in the survival ability of the probiotic cultures in the two yoghurt types. L. acidophilus LAI demonstrated, in general, a lower resistance to the yoghurt environment than B. bifidum BBI. On the other hand, the full-fat yoghurt was a more inhibitory medium than the reduced-fat one, especially for B. bifidum BBI. Regarding the lactic starters used, the results showed that the culture SISD was clearly more inhibitory for both probiotic organisms than the culture SID. The loss of cell viability in yoghurt samples was different (higher in some cases and lower in others) from that due to lactic acid only. In general, pH values of 4.5 or lower jeopardised the cell viability of the probiotic organisms in yoghurt stored at 5°C. This work shows the importance of selecting a suitable combination of probiotic strains and starter cultures when different yoghurt types are formulated.     

Selenium enriched green tea increase stability of Lactobacillus casei and Lactobacillus plantarum in chitosan coated alginate microcapsules during exposure to simulated gastrointestinal and refrigerated conditions

The effects of selenium enriched green tea (SGT; 85.8–96 mg/kg) in different concentrations of 1 g and 2 g/100 mL, on the in vitro exposure to simulated gastrointestinal juice and refrigerated storage of encapsulated Lactobacillus casei and Lactobacillus plantarum were investigated in chitosan coated alginate beads. The encapsulation yield of viable cells in chitosan coated alginate beads with and without SGT was not significantly different (P < 0.05). These results together with the study about the survival of probiotic bacteria in microspheres with SGT during storage at 4 °C, demonstrated significantly higher number (P < 0.05) of survival bacteria in microcapsules with SGT 2 g/100 mL. Microencapsulated L. casei and L. plantarum with SGT 1 g and 2 g/100 mL were resistant to simulated gastric conditions (pH 2.0, 2 h) and bile solution (3 g/100 mL, 2 h) resulting in significantly (P < 0.05) improved survival when compared with microencapsulation without SGT addition.     

Microencapsulation of Lactobacillus reuteri DSM 17938 Cells Coated in Alginate Beads with Chitosan by Spray Drying to Use as a Probiotic Cell in a Chocolate Soufflé

The main objective of this work was to obtain microencapsulated probiotic cells in order to improve their resistance to heat stress and gastrointestinal conditions. A further aim was to obtain a potentially probiotic chocolate soufflé. Lactobacillus reuteri DSM 17938 cells were microencapsulated by spray drying in alginate matrix and further coated with chitosan. Bacterial survival after exposure to different heat treatments and simulated gastrointestinal conditions were measured to test the microcapsules. They were also dyed by using a LIVE/DEAD^® BacLight? Bacterial Viability Kit and characterized by epifluorescence microscope observation. Furthermore, a potentially chocolate soufflé was prepared using microencapsulated cells. The results indicated that alginate microcapsules did not improve acid tolerance or heat resistance in “in vitro” experiments, while they were able to protect 7% of the Lactobacillus reuteri population during the baking of a chocolate soufflé, compared to a survival rate of 1% of free cells. By contrast, the cells microencapsulated with alginate coated with chitosan showed, compared to free cells, improved acid tolerance, allowing the cell population to remain constant after 3 h in simulated gastric conditions. Moreover, the heat resistance of cells in co-cross-linked microcapsules significantly improved compared to free cells, both in “in vitro” and “in food” experiments. Microencapsulation led to a survival rate of 10% after baking a chocolate soufflé. However, the final level of bacterial cells in the product was too low to consider the chocolate soufflé as a probiotic product.     

Comparison of vacuum impregnation and soaking techniques for addition of the probiotic Lactobacillus acidophilus to minimally processed melon

This study aimed to evaluate the vacuum impregnation (VI) and soaking methods in the addition of Lactobacillus acidophilusLA‐3 to minimally processed melon (MPM). The melons were washed, sanitised in chlorine solution (200 mg L^?1), peeled and cutted into cubes. Lactobacillus acidophilusLA‐3 (1.4 × 10¹⁰ CFU g^?1) were added to the MPM through both techniques. The L. acidophilusLA‐3 count in MPM was similar to those commonly found in dairy products having probiotic claim, but VI was more efficient than soaking in maintaining the viability (8.61 and 7.98 Log CFU g^?1, respectively). The pH, acidity and soluble solids were not affected by probiotic culture and the incorporation technique; however, the VI affected the firmness of fruit. The MPM was within Brazilian standards for their microbiological characteristics. MPM may be used as a carrier of probiotic bacteria, being one more alternative for individuals who consume probiotic products.     

Application of shellac for the development of probiotic formulations

In this study, we have improved the enteric properties of shellac and developed probiotic formulations comprising this natural polymer. The effects of plasticizers such as glycerol and glyceryl triacetate, as well as water-soluble polymers such as sodium alginate, hydroxypropyl methylcellulose and polyvinylpyrrolidone on thermodynamic characteristics and coating properties of shellac were evaluated. The data indicate that glycerol showed the best plasticization effect. Hydroxypropyl methylcellulose and polyvinylpyrrolidone had superior miscibility with shellac compared to sodium alginate. Then, three fluid-bed dried bacterial species i.e., Enterococcus faecium, Bifidobacterium bifidum and Lactobacillus reuteri, were coated with formulations comprising different concentrations of shellac and additives. Coatings with shellac containing 5% glycerol or 5% sodium alginate or up to 20% [w/w] polyvinylpyrrolidone protected the microorganisms against acidic pH and provided the best release profile in simulated intestinal fluid. Moreover, these formulations maintained promising cell survival rates after four months of storage at 5 °C. E. faecium and B. bifidum showed more resistance to manufacturing process than L. reuteri.     

Encapsulation of probiotic Lactobacillus acidophilus by ionic gelation with electrostatic extrusion for enhancement of survival under simulated gastric conditions and during refrigerated storage

The probiotic Lactobacillus acidophilus was encapsulated in biodegradable and biocompatible capsules prepared by ionic gelation between phytic acid (PA) and chitosan (CS) with an electrostatic extrusion method. Calcium carbonate (CaCO₃) and starch were used as co‐encapsulants for improvement of capsule stability. Capsules were characterised and evaluated for survival of encapsulated L. acidophilus cells in simulated gastric fluid (SGF) and during refrigerated storage. Loading capacity values of PA‐CS capsules, PA‐CS‐starch capsules and PA‐CS‐CaCO₃ capsules were 8.20, 8.12 and 7.81 log CFU g^?1 of wet capsule, respectively. Capsules showed particle sizes of 1.3–1.5 mm and a uniform spherical shape. PA‐CS‐CaCO₃ capsules were the most stable vehicles for the protection of probiotic cells against acidic damage, particularly at pH 1.5 and pH 2. L. acidophilus cells from PA‐CS‐CaCO₃ capsules showed only a 0.64 log CFU reduction in numbers after 2 h in pH 1.5 SGF conditions. The numbers of L. acidophilus encapsulated in PA‐CS‐CaCO₃ capsules were decreased by only 0.69 log CFU g^?1, while PA‐CS capsules and PA‐CS‐starch capsule numbers were reduced by more than 1.45 log CFU g^?1 after 4 weeks at 4 °C. Addition of calcium carbonate to PA‐CS capsules provided protection against acid injury via antacid and buffering effects for encapsulation of L. acidophilus.     

Fermentation of beet juice by beneficial lactic acid bacteria

Red beets were evaluated as a potential substrate for the production of probiotic beet juice by four species of lactic acid bacteria (Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus plantarum). All the lactic cultures were found capable of rapidly utilizing beet juice for cell synthesis and lactic acid production. However, L. acidophilus and L. plantarum produced a greater amount of lactic acid than other cultures and reduced the pH of fermented beet juice from an initial value of 6.3 to below 4.5 after 48 h of fermentation at 30°C. Although the lactic cultures in fermented beet juice gradually lost their viability during cold storage, the viable cell counts of these lactic acid bacteria except for L. acidophilus in the fermented beet juice still remained at 10⁶–10⁸ CFU/ml after 4 weeks of cold storage at 4°C.     

Characteristics of carbonated fermented milk and survival of probiotic bacteria

The carbonation of pasteurised milk was evaluated as a method for improving bacterial viability in fermented milk added with probiotic bacteria (Lactobacillus acidophilus and/or Bifidobacterium bifidum). The behaviour of microorganisms during fermentation and cold storage, and the biochemical and sensory properties of the products were assessed. In AT (Streptococcus thermophilus/L. acidophilus) and ABT (S. thermophilus/L. acidophilus/B. bifidum) products, the fermentation times to decrease the pH to 5 were significantly lowered when CO₂ or lactic acid was added to milk. The higher acidity levels of carbonated (as a result of production of carbonic acid) and lactic acidified samples enhanced growth and metabolic activity of the starter during fermentation and was the reason for this reduction in incubation time. Cell counts of S. thermophilus, L. acidophilus and B. bifidum gradually decreased through the cold storage of carbonated and non-acidified fermented milk, although the counts were always higher than 10⁶ viable cells g⁻¹. The CO₂ did not exert any influence on the viability of S. thermophilus and L. acidophilus in AT fermented milk stored at 4°C but the presence of B. bifidum and CO₂ in ABT-type products was associated with lower viability of L. acidophilus during the refrigerated storage. The higher acetate concentrations of ABT products made with non-acidified milk as compared with the carbonated products could have contributed to major survival of L. acidophilus in the former. The use of milk acidified with CO₂ had no detrimental effects on the sensory properties of ABT fermented milk. Therefore, we concluded that the carbonation of pasteurised milk prior to the starter addition could be satisfactorily used to reduce the manufacture time of fermented milk.     

Preparation of Acid‐Resistant Microcapsules with Shell‐Matrix Structure to Enhance Stability of Streptococcus Thermophilus IFFI 6038

Microencapsulation is an effective technology used to protect probiotics against harsh conditions. Extrusion is a commonly used microencapsulation method utilized to prepare probiotics microcapsules that is regarded as economical and simple to operate. This research aims to prepare acid‐resistant probiotic microcapsules with high viability after freeze‐drying and optimized storage stability. Streptococcus thermophilus IFFI 6038 (IFFI 6038) cells were mixed with trehalose and alginate to fabricate microcapsules using extrusion. These capsules were subsequently coated with chitosan to obtain chitosan‐trehalose‐alginate microcapsules with shell‐matrix structure. Chitosan‐alginate microcapsules (without trehalose) were also prepared using the same method. The characteristics of the microcapsules were observed by measuring the freeze‐dried viability, acid resistance, and long‐term storage stability of the cells. The viable count of IFFI 6038 in the chitosan‐trehalose‐alginate microcapsules was 8.34 ± 0.30 log CFU g^?1 after freeze‐drying (lyophilization), which was nearly 1 log units g^?1 greater than the chitosan‐alginate microcapsules. The viability of IFFI 6038 in the chitosan‐trehalose‐alginate microcapsules was 6.45 ± 0.09 log CFU g^?1 after 120 min of treatment in simulated gastric juices, while the chitosan‐alginate microcapsules only measured 4.82 ± 0.22 log CFU g^?1. The results of the long‐term storage stability assay indicated that the viability of IFFI 6038 in chitosan‐trehalose‐alginate microcapsules was higher than in chitosan‐alginate microcapsules after storage at 25 °C. Trehalose played an important role in the stability of IFFI 6038 during storage. The novel shell‐matrix chitosan‐trehalose‐alginate microcapsules showed optimal stability and acid resistance, demonstrating their potential as a delivery vehicle to transport probiotics.