Abstract
Purpose: To optimize the process parameters for the encapsulation of Clostridium tyrobutyricum (Ct) and to determine its in vitro characteristics.Methods: The process parameters, including the concentration of the wall and hardening material, Ct to gelatin ratio and hardening time, were studied by single factor analysis, while optimization was performed by orthogonal experimental design for the encapsulation rate of Ct.Results: Optimal conditions exhibited by orthogonal experimental design at a 92.17 % encapsulation rate with a viable count of 9.61 ± 0.06 lgCFU/g were: 6 % modified starch, 3 % sodium alginate, and 2 % CaCl2 at a Ct to gelatin ratio of 1:1 with a hardening time of 30 min. The survival rates of encapsulated Ct were higher than free Ct in simulated gastric (6.22 %) and intestinal juices (15.55 %). Reduction in viable counts of Ct at 90 °C were higher for free cells (44.76 %) than encapsulated cells (28.09 %) after 30 min of heat treatment. Correspondingly, encapsulation boosted the capacity of Ct to withstand the strong acidic conditions of the stomach and improved the storage properties of Ct.Conclusion: The results suggested that extrusion is a good technique for the encapsulation of Ct, as it enhances the viability of Ct during their transit through the gastrointestinal tract. Furthermore, encapsulation is favorable for Ct if planned for use in formulations where high temperature treatment is required.
Highlights
Probiotics are live microorganisms that have beneficial effects on the host when fed in adequate amounts
The best combination of parameters was 6 % M-starch, 3 % Na-alginate, 2 % CaCl2, and Clostridium tyrobutyricum (Ct) to gelatin ratio of 1:1 with a hardening time of 30 min, which were validated by a follow-up experiment, and the highest microencapsulation rate (ME rate) (92.17 %) and viable counts (9.61 ± 0.06 LgCFU/g) were observed at these selected parameters compared to 16 combinations studied previously (Table 2)
The results of the 0.3% bile salt treatment revealed that ME had no effect against bile salt tolerance (Figure 6)
Summary
Probiotics are live microorganisms that have beneficial effects on the host when fed in adequate amounts. Probiotics have many physiological functions, such as killing harmful bacteria, improving digestion and absorption, and enhancing growth, productivity and intestinal immune function [1], there are many problems associated with their application as feed additives These include low survival of live bacteria during processing and. Encapsulation of lactic acid bacteria using calcium alginate as a wall material significantly improves heat resistance and gastrointestinal survival [3]. Encapsulation significantly improves the survival of probiotics in different situations, huge variation in the concentration and type of wall materials and lack of consideration of all parameters make it difficult to select the best parameters. This study was designed to optimize the process parameters for microencapsulation of Ct at maximum rate with more viable bacteria and in vitro characterization of prepared microcapsules under different conditions. Single factor analysis of the Ct to gelatin ratio showed that 1:3 was the best ratio for the peak ME rate (78.28 %) under the conditions of this study (Figure 2 e)
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