Effect of Type of Protein‐Based Microcapsules and Storage at Various Ambient Temperatures on the Survival and Heat Tolerance of Spray Dried Lactobacillus acidophilus

The aims of this study were to evaluate the effect of types of protein‐based microcapsules and storage at various ambient temperatures on the survival of Lactobacillus acidophilus during exposure to simulated gastrointestinal tract and on the change in thermo‐tolerance during heating treatment. The encapsulating materials were prepared using emulsions of protein (sodium caseinate, soy protein isolate, or pea protein), vegetable oil, and glucose, with maltodextrin was used as a wall material. The formulations were heated at 90 °C for 30 min to develop Maillard substances prior to being incorporated with L. acidophilus. The mixtures were then spray dried. The microspheres were stored at 25, 30, and 35 °C for 8 wk and examined every 4 wk. The addition of proteins as encapsulating materials demonstrated a significant protective effect (P < 0.05) as compared to the control sample. Sodium caseinate and soy protein isolate appeared more effective than pea protein in protecting the bacteria after spray drying and during the storage at different room temperatures. Storage at 35 °C resulted in a significant decrease in survival at end of storage period regardless the type of encapsulating materials. The addition of protein‐based materials also enhanced the survival of L. acidophilus during exposure to simulated gastrointestinal condition as compared to the control. After spray drying and after 0th wk storage, casein, soy protein isolate, and pea protein‐based formulations protected the bacteria during heat treatment. In fact, a significant decrease in thermal tolerance was inevitable after 2 wk of storage at 25 °C.     

Stability of microencapsulated Lactobacillus acidophilus and Lactococcus lactis ssp. cremoris during storage at room temperature at low aw

The effect of freeze drying or spray drying, the use of desiccants to maintain the low a_w and the period of storage (at 25 °C) of Lactobacillus acidophilus and Lactococcus lactis ssp. cremoris on survival, acid tolerance, bile tolerance, retention of surface hydrophobicity and retention of β-galactosidase was studied; an estimation of the maximum storage period was also carried out. Sodium caseinate, vegetable oil, glucose, mannitol and fructooligosaccharides were used as protectant of L. acidophilus and L. cremoris during freeze drying or spray drying and during subsequent storage. NaOH, LiCl and silica gel were used as desiccants during 10 weeks of storage of microencapsulated L. acidophilus and L. cremoris kept in an aluminum foil pouch. The results showed that mainly freeze dried L. acidophilus and L. cremoris kept in foil pouch containing NaOH (a_w 0.07) or LiCl (a_w 0.1) showed higher survival (89–94%) than spray dried bacteria kept under the same conditions (86–90%) after 10 weeks of storage (P = 0.0005). Similar results were also showed by acid tolerance, bile tolerance and surface hydrophobicity of freeze-dried or spray-dried L. acidophilus and L. cremoris. Silica gel was less effective in protecting the functional properties of microencapsulated L. acidophilus or L. cremoris with percentage of survival between 81 and 87% at week 10 of the storage. However, retention of β-galactosidase was only influenced by a_w adjusted by desiccators (P < 0.05). Based on forecasting using linear regression, the predicted storage period for freeze dried L. acidophilus, spray dried L. acidophilus and freeze dried L. cremoris kept in foil pouch containing NaOH would be 46, 42 and 42 weeks, respectively; while spray dried L. cremoris under LiCl desiccant would require 39 weeks to achieve minimum required bacterial population of 10⁷ CFU/g.     

Survival, acid and bile tolerance, and surface hydrophobicity of microencapsulated B. animalis ssp. lactis Bb12 during storage at room temperature

Survival, acid and bile tolerance, and surface hydrophobicity of microencapsulated Bifidobacterium animalis ssp. lactis Bb12 were studied during storage at room temperature (25 °C) at low water activity (0.07, 0.1, and 0.2). Two types of alginate-based systems were prepared with and without mannitol as microencapsulant of B. animalis ssp. lactis Bb12. Formation of gel beads containing cells was achieved by dropping each emulsion into CaCl(2) solution; then, the beads were freeze dried. Survival, acid tolerance during 2-h exposure in de Man, Rogosa, Sharpe (MRS) broth at pH 2.0, bile tolerance during 8-h exposure in MRS broth containing taurocholic acid at pH 5.8, and retention of surface hydrophobicity were determined after freeze drying and during storage. The result showed that neither alginate nor alginate-mannitol formulation was effective in protecting B. animalis ssp. lactis Bb12 during freezing and freeze drying. The viability in alginate-mannitol and alginate formulations after freeze drying was 6.61 and 6.34 log CFU/g, respectively. Storage at low a(w) improved survival, acid tolerance, bile tolerance, and surface hydrophobicity retention of microencapsulated B. animalis ssp. lactis Bb12 when compared with controlled storage in an aluminum foil (with a(w) of 0.38 and 0.40 for alginate-mannitol and alginate formulations, respectively). Alginate mannitol was more effective than the alginate system during a short period of storage, but its effectiveness decreased during a long period of storage (80% survival at 10 wk). Nevertheless, storage of microencapsulated B. animalis ssp. lactis Bb12 in an aluminum foil without a(w) adjustment during 10 wk at room temperature was not effective (survival was 64% to 65%).     

Survival of Microencapsulated Probiotic Bacteria after Processing and during Storage: A Review

The use of live probiotic bacteria as food supplement has become popular. Capability of probiotic bacteria to be kept at room temperature becomes necessary for customer's convenience and manufacturer's cost reduction. Hence, production of dried form of probiotic bacteria is important. Two common drying methods commonly used for microencapsulation are freeze drying and spray drying. In spite of their benefits, both methods have adverse effects on cell membrane integrity and protein structures resulting in decrease in bacterial viability. Microencapsulation of probiotic bacteria has been a promising technology to ensure bacterial stability during the drying process and to preserve their viability during storage without significantly losing their functional properties such acid tolerance, bile tolerance, surface hydrophobicity, and enzyme activities. Storage at room temperatures instead of freezing or low temperature storage is preferable for minimizing costs of handling, transportation, and storage. Concepts of water activity and glass transition become important in terms of determination of bacterial survival during the storage. The effectiveness of microencapsulation is also affected by microcapsule materials. Carbohydrate- and protein-based microencapsulants and their combination are discussed in terms of their protecting effect on probiotic bacteria during dehydration, during exposure to harsh gastrointestinal transit and small intestine transit and during storage.     

Viability,Acid and Bile Tolerance of Spray Dried Probiotic Bacteria and Some Commercial Probiotic Supplement Products Kept at Room Temperature

Production of probiotic food supplements that are shelf‐stable at room temperature has been developed for consumer's convenience, but information on the stability in acid and bile environment is still scarce. Viability and acid and bile tolerance of microencapsulated Bifidobacterium spp. and Lactobacillus acidophilus and 4 commercial probiotic supplements were evaluated. Bifidobacterium and L. acidophilus were encapsulated with casein‐based emulsion using spray drying. Water activity (a_w) of the microspheres containing Bifidobacterium or L. acidophilus (SD GM product) was adjusted to 0.07 followed by storage at 25 °C for 10 wk. Encapsulated Bifidobacterium spp. and Lactobacillus acidophilus and 4 commercial probiotic supplement products (AL, GH, RE, and BM) were tested. Since commercial probiotic products contained mixed bacteria, selective media MRS‐LP (containing L‐cysteine and Na‐propionate) and MRS‐clindamycin agar were used to grow Bifidobacterium spp. or L. acidophilus, respectively, and to inhibit the growth of other strains. The results showed that a_w had a strong negative correlation with the viability of dehydrated probiotics of the 6 products. Viable counts of Bifidobacterium spp. and L. acidophilus of SD GM, AL, and GH were between 8.3 and 9.2 log CFU/g, whereas that of BM and RE were between 6.7 and 7.3 log CFU/g. Bifidobacterium in SD GM, in AL, and in GH products and L. acidophilus in SD GM, in AL, and in BM products demonstrated high tolerance to acid. Most of dehydrated probiotic bacteria were able to survive in bile environment except L. acidophilus in RE product. Exposure to gastric juice influenced bacterial survivability in subsequent bile environment.     

Enzyme stability of microencapsulated Bifidobacterium animalis ssp. lactis Bb12 after freeze drying and during storage in low water activity at room temperature

Abstract: Stability of enzymes such as β‐galactosidase (β‐gal), β‐glucosidase (β‐glu), lactate dehydrogenase (LDH), pyruvate kinase (PK), hexokinase (HK), and ATPase of microencapsulated Bifidobacterium animalis ssp. lactis Bb12 after freeze‐drying and after 10 wk of storage at low water activity (a_w) at room temperature was studied. Bacteria were microencapsulated using alginate formulation with or without mannitol fortification (sodium alginate and mannitol [SAM] and sodium alginate [SA], respectively) by creating gel beads followed by freeze drying. Two types of dried gel beads were then stored at low a_w, such as 0.07, 0.1, and 0.2; storage in an aluminum foil was used as control. All storage was carried out at room temperature of 25 °C for 10 wk. Measurement of β‐gal, β‐glu, LDH, PK, HK, and ATPase (with or without exposure to pH 2.0 for 2 h) activities was carried out before freeze drying, after freeze drying, and after 10 wk of storage. There was a significant decrease in almost all enzyme activities, except that of PK. SAM and SA showed no different effect on maintaining enzyme activities during freeze drying. Storage for 10 wk at room temperature at various low a_w using SAM and SA system had a significant effect on retention of most enzymes studied, except that of PK and LDH. Storage at a_w of 0.07 and 0.1 was more effective in maintaining enzyme activities than storage at a_w of 0.2 and in an aluminum foil. However, mannitol fortification into alginate system did not significantly improve retention of enzymes during 10 wk of storage.