Improved production of lactiplantibacillus plantarum RO30 exopolysaccharide (REPS) by optimization of process parameters through statistical experimental designs.

BACKGROUND: In investigating of (exopolysaccharide) EPS from unconventional sources, lactic acid bacteria have a vital role due to their generally recognized as safe (GRAS) status. EPSs have diverse applications such as drug delivery, antimicrobial activity, surgical implants, and many more in many sectors. Despite being important, the main hindrance to the commercial application of these significant biopolymers is low productivity. Therefore, this study primarily focuses on optimizing physio-chemical conditions to maximize the previously produced EPS from probiotic Lactiplantibacillus plantarum RO30 (L. plantarum RO30) using one factor at a time (OFAT) and method Response Surface Methodology (RSM). RESULTS: The EPS obtained from L. plantarum RO30 named REPS. The medium formulation for REPS production using the OFAT method revealed that sucrose (20 g/L, beef extract (25 g/L), and ammonium sulfate at 4 g/L concentration were the optimum carbon, organic and inorganic nitrogen sources, and REPS yield was increased up to 9.11 ± 0.51 g/L. RSM experiments revealed that, a greatly significant quadratic polynomial attained from the Central Composite Design (CCD) model was fruitful for specifying the most favorable cultural conditions that have significant consequences on REPS yield. The maximal amount of REPS (10.32 g/L) was formed by: sucrose (40 g/L), beef extract (25 g/L), pH (5.5), incubation temperature (30 °C), and incubation period (72 h). A high closeness was obtained between the predicted and experimental values and it displayed the efficiency of the RSM. CONCLUSION: This study was conducted to reinforce REPS production in the probiotic LAB L. plantarum RO30 by utilizing various experimental parameters. The maximum REPS yield of 10.32 g/L was attained under the circumstances optimized in the study.

Exopolysaccharide produced by Lactiplantibacillus plantarum RO30 isolated from Romi cheese: characterization, antioxidant and burn healing activity.

Microbial exopolysaccharides (EPSs) extracted from lactic acid bacteria (LAB) are generally recognized as safe. They have earned popularity in recent years because of their exceptional biological features. Therefore, the present study main focus was to study EPS-production from probiotic LAB and to investigate their antioxidant and burn wound healing efficacy. Seventeen LAB were isolated from different food samples. All of them showed EPS-producing abilities ranging from 1.75 ± 0.05 to 4.32 ± 0.12 g/l. RO30 isolate (from Romi cheese) was chosen, due to its ability to produce the highest EPS yield (4.23 ± 0.12 g/l). The 16S rDNA sequencing showed it belonged to the Lactiplantibacillus plantarum group and was further identified as L. plantarum RO30 with accession number OL757866. It displayed well in vitro probiotic properties. REPS was extracted and characterized. The existence of COO-, OH and amide groups corresponding to typical EPSs was confirmed via FTIR. It was constituted of glucuronic acid, mannose, glucose, and arabinose in a molar ratio of 2.2:0.1:0.5:0.1, respectively. The average molecular weight was 4.96 × 104 g/mol. In vitro antioxidant assays showed that the REPS possesses a DPPH radical scavenging ability of 43.60% at 5 mg/ml, reducing power of 1.108 at 10 mg/ml, and iron chelation activity of 72.49% and 89.78% at 5 mg/ml and 10 mg/ml, respectively. The healing efficacy of REPS on burn wound models in albino Wistar rats showed that REPS at 0.5% (w/w) concentration stimulated the process of healing in burn areas. The results suggested that REPS might be useful as a burn wound healing agent.

Survivability of alginate-microencapsulated Lactobacillus plantarum during storage, simulated food processing and gastrointestinal conditions.

A comparison between the most investigated alginate-based encapsulating agents was performed in the current study. Here, the survivability of Lactobacillus plantarum microencapsulated with alginate (Alg) combined with skim milk (Sm), dextrin (Dex), denatured whey protein (DWP) or coated with chitosan (Ch) was evaluated after exposure to different heat treatments and in presence of some food additives, during storage and under simulated gastrointestinal condition. In addition, the encapsulated cells were evaluated for production of different bioactive compounds such as exopolysacchar. ides and antimicrobial substances compared with the unencapsulated cells. The results showed that only Alg-Sm maintained the viability of the cells >106 cfu/g at the pasteurization temperature (65 °C for 30 min). Interestingly, storage under refrigeration conditions increased the viability of L. plantarum entrapped within all the tested encapsulating agents for 4 weeks. However, under freezing condition, only Alg-DWP and Alg-Sm enhanced the survival of the entrapped cells for 3 months. All the microencapsulated cells were capable of growing at the different NaCl concentrations (1%-5%) except for cells encapsulated with Alg-Dex, showed viability loss at 3% and 5% NaCl concentrations. Tolerance of the microencapsulated cells toward organic acids was varied depending on the type of organic acid. Alg-Ch and Alg-Sm provide better survival for the cells under simulated gastric juice; however, all offer a good survival for the cells under simulated intestinal condition. Our findings indicated that Alg-Sm proved to be the most promising encapsulating combination that maintains the survivability of L. plantarum to the recommended dose level under almost all the stress conditions adopted in the current study. Interestingly, the results also revealed that microencapsulation does not affect the metabolic activity of the entrapped cells and there was no significant difference in production of bioactive compounds between the encapsulated and the unencapsulated cells.