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. 2024 Aug 9;14(1):18518.
doi: 10.1038/s41598-024-68086-3.

In vitro digestive system simulation and anticancer activity of soymilk fermented by probiotics and synbiotics immobilised on agro-industrial residues

Abdallah I Gad  1 , Mona M Orabi  1 , Khadiga A Abou-Taleb  2 , Dina Y Abdelghani  3 , Shimaa A Amin  1
Affiliations

In vitro digestive system simulation and anticancer activity of soymilk fermented by probiotics and synbiotics immobilised on agro-industrial residues

Abdallah I Gad et al. Sci Rep. .

Abstract

In this study, a variety of probiotic strains, including Lactiplantibacillus plantarum, Lacticaseibacillus casei, Lactobacillus acidophilus, Streptococcus thermophilus, Bifidobacterium longum, Limosilactobacillus reuteri, Lactobacillus delbrueckii subsp. bulgaricus, Lacticaseibacillus rhamnosus, and Bifidobacterium bifidum, were utilized for soymilk fermentation both as free cells and as synbiotics on agro-industrial residuals such as okara, whey protein, banana peels, apple pomace, sugarcane bagasse, orange peels, and lemon peels. Among these, Lacticaseibacillus rhamnosus emerged as the most significant strain for soymilk fermentation, exhibiting a viability of 10.47 log cfu/mL, a pH of 4.41, total acidity of 1.12%, and organic acid contents (lactic and acetic acid) of 11.20 and 7.50 g/L, respectively. As a synbiotic Lacticaseibacillus rhamnosus immobilised on okara, showed even more impressive results, with a viability of 12.98 log cfu/mL, a pH of 4.31, total acidity of 1.27%, and organic acid contents of 13.90 and 9.30 g/L, respectively. Over a 12-h fermentation period, cell viability values increased by 10.47-fold in free cells and 11.19-fold in synbiotics. Synbiotic supplementation of fermented soymilk proved more beneficial than free cells in terms of viability, acidity, and organic acid content. Furthermore, when synbiotic fermented soymilk was freeze-dried to simulate the digestive system in vitro, synbiotics and freeze-dried cells demonstrated superior gastrointestinal tract survival compared to free cells. Both the probiotic bacteria and the synbiotics exhibited cytotoxicity against colon and liver cancer cell lines, with half-maximal inhibitory concentrations ranging from 41.96 to 61.52 μL/well.

Keywords: Agro-industrial wastes; And anticancer; Cytotoxicity; Encapsulation; Freeze-dried; Gastrointestinal track; Probiotic bacteria and synbiotic; Soymilk.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
The change in organic acids content of fermented soymilk by free probiotic bacterial strains affected with fermentation time. Lactiplantibacillus plantarum ATCC 14,917 (S1), Lactiplantibacillus plantarum DSA 20,174 (S2), Lacticaseibacillus caseiDSM 20,011 (S3), Limosilactobacillus reuteri NRRL B-14171 ((S4), Lactobacillus acidophilus ATCC 20,552 (S5),lactobacillus delbrueckii sub sp. bulgaricus DSMZ 20,080 (S6), Lacticaseibacillus rhamnosus NRRL B-442 (S7), Streptococcus thermophilus DSM 20,259 (S8), Bifidobacterium bifidum NRRL B-41410 (S9), and Bifidobacterium longum B41409 (S10).
Figure 2
Figure 2
Organic acids (lactic and acetic or propionic acids) productivity during soymilk fermentation by free probiotic bacterial strains affected with fermentation time. Lactiplantibacillus plantarum ATCC 14,917 (S1), Lactiplantibacillus plantarum DSA 20,174 (S2), Lacticaseibacillus caseiDSM 20,011 (S3), Limosilactobacillus reuteri NRRL B-14171 ((S4), Lactobacillus acidophilus ATCC 20,552 (S5),lactobacillus delbrueckii sub sp. bulgaricus DSMZ 20,080 (S6), Lacticaseibacillus rhamnosus NRRL B-442 (S7), Streptococcus thermophilus DSM 20,259 (S8), Bifidobacterium bifidum NRRL B-41410 (S9), and Bifidobacterium longum B41409 (S10).
Figure 3
Figure 3
Effect of acidic condition (A) and bile salts (B) on survival of free cells, synbiotic, encapsulated Lacticaseibacillus rhamnosus NRRL B-442. FSMFC, fermented soymilk by free Lacticaseibacillus rhamnosus NRRL B-442.
Figure 4
Figure 4
In Vitro Survival of free Lacticaseibacillus rhamnosus NRRL B-442 cells, synbiotic and encapsulated under digestion process conditions (salivary, gastric and intestinal phases). (A) Viability (log cfu/mL), (B) Percentage survival rate and (C) Percentage of inhibition rate.
Figure 5
Figure 5
Representative photomicrograph of HCT-116, and HepG2 cells morphology before treated (A), and after treated by soymilk extracts fermented by free (B), and symbiotic (C), photographed with an inverted phase-contrast microscope (magnification 100 ×).
See this image and copyright information in PMC

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