Survival enhancement of probiotic bacteria by encapsulation with chitosan succinate nano-particles and evaluation of survival in simulated gastrointestinal conditions

Abstract

The present study describes chitosan succinate (as nano particals) to protect Bifidobacterium lactis Bb-12 from simulated gastric conditions (SGC) . Chitosan succinate polymer was synthesized and emulsification to prepare microspheres. Besides, the microspheres of Bifidobacterium lactis BB-12 and chitosan succinate particle size were determined by scanning electron microscopy, FT-IR spectroscopy.

The characteristic and the results of absorption peaks of chitosan succinate microspheres by used IR spectra was 1495 cm_1 and degree of electrostatic repulsion of chitosan succinate particle in the microspheres was -33 m using Zeta potential measurement, and chitosan succinate microspheres had a net negative charge. Moreover, that the size of chitosan succinate microspheres was chitosan succinate microspheres 482.8 nm. The width diameter of free cells was 2.144 µm (2144 nm), while the width diameter encapsulation of the microspher with and without Genipin were 2.925 µm (2925 nm) and 2.212 µm (2212nm), respectively were determined by Zeta potential

In vitro release chitosan succinate microspheres with Genipin croos-link were had protect effect of the degradation from gastric enzymes than Nanoencapsulation cell (NEC) without Genipin. The encapsulated Bb-12 was significant highest cells survival in simulated intestinal juice in pH 8 and simulated colon juice in pH 7.4, while the encapsulated BB-12 had low number of survival was released in simulated gastric juice (pH 2.0).

pH can be effect of release matrix form in chitosan succinate microsphere. Chitosan succinate microsphere cross-link with Genipin had significanly high level of survival which was observed under pH 7.4 (Colon) However, this can be resist in Gasteric conditions than chitosan succinate microsphere no cross-link with Genipin.

The stability of chitosan succinate cross-linked with Genipin was observed at a high stability in the acidic environment of simulated gastric juice with absent of pepsin enzyme (2.60±3.60×105a in initial time of incubation). Furthermore, the simulated gastric juice with pepsin enzyme had reduced stability significantly (3.20±2.50×103a after 90 min of incubation). The stability of chitosan succinate without cross-linked with genipin observed that there was no significant effect (4.21±2.20×104d and 4.90±4.72×104a), of simulated gastric juice with or without pepsin enzyme. While, the survival of free cell was reduced to <10 cfu/ml after 30 min of incubation gastric juice without pepsin enzyme and to 3.44±2.70×102c after 120 min of incubation in gastric juice with pepsin.

Genipin under concentration 2.5 mM cross-link might be enhance the resistance of chitosan succinate to degradation by pepsin enzyme for controlled release of probiotic cells. According to these results the new carrier system described can be a promising tool probiotic delivery from oral to colon.

Recently, foods with functional properties are needed. Tomato ketchup has high contents of total sugar and consumed in many fast food. The objective of this study was to monitor the growth and survival of probiotic bacteria in Bifidobacetrium lactis Bb-12 strain in local tomato ketchup. The NEC Bifidobacetrium lactis Bb-12 count increased gradually in tomato Ketchup up to 2 days of incubation. While, the count of free cell was decreased after 90 min of incubation.

However, the count of NEC of Bb-12 was increased significantly after initial time to tow days and then decreased to not count after 7 days of incubation time. While, the survival of free cell was increased until 90 min then decreased to <10 cfu/ml after 2 days of incubation in tomato ketchup. However, an inverse relationship was observed between the time of incubation and the survival rate of free Bb-12 cells. The study results showed a good protection activity of NEC of Bb-12 for rigors tomato Ketchup for 2 days.

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