# Formulation and in-vitro functional evaluation of a Bacillus-based multi-strain probiotic consortium relevant to protein-energy malnutrition

**Authors:** Priya Mori, Ishita Modasiya, Mehul Chauhan, Hina Maniya, Vijay Kumar, Apurba Kumar Sarkar, Babak Pakbin, Babak Pakbin, Babak Pakbin, Babak Pakbin

PMC · DOI: 10.1371/journal.pone.0345821 · PLOS One · 2026-03-24

## TL;DR

This study develops a multi-strain Bacillus probiotic that improves protein digestion and reduces gut inflammation in protein-energy malnutrition.

## Contribution

A novel Bacillus-based multi-strain probiotic consortium is formulated and shown to outperform single strains in addressing PEM.

## Key findings

- The probiotic consortium showed high proteolytic activity (0.52 ± 0.03 U/mL).
- It exhibited strong anti-inflammatory potential (5.33 ± 0.06 U/mL).
- The consortium tolerated gastrointestinal conditions and had antioxidant properties.

## Abstract

Protein-energy malnutrition (PEM) remains a critical global health challenge, characterized by impaired nutrient absorption and chronic gut inflammation. While probiotics offer a potential therapeutic avenue, the efficacy of single-strain interventions is often limited. This study aimed to formulate and evaluate a Bacillus-based multi-strain probiotic consortium (MSPC) specifically tailored for PEM. Three strains—Bacillus spizizenii BAB 7915, Bacillus tequilensis, and Bacillus rugosus-were selected based on their non-antagonistic, synergistic growth profiles. The MSPC demonstrated superior functional attributes compared to individual strains, exhibiting significant proteolytic activity (0.52 ± 0.03 U/mL) and robust anti-inflammatory potential (5.33 ± 0.06 U/mL). Additionally, the consortium showed high tolerance to gastrointestinal conditions and strong antioxidant properties. These results suggest that the MSPC can effectively enhance protein hydrolysis and mitigate gut inflammation, providing a scientifically validated, low-cost formulation for the nutritional rehabilitation of PEM patients.

## Linked entities

- **Diseases:** Protein-energy malnutrition (MONDO:0001371)
- **Species:** Bacillus spizizenii (taxon 96241), Bacillus tequilensis (taxon 227866), Bacillus rugosus (taxon 2715209)

## Full-text entities

- **Genes:** ALB (albumin) [NCBI Gene 280717]
- **Diseases:** wasting (MESH:D019282), Overgrowth (MESH:C537340), Malnutrition (MESH:D044342), obesity (MESH:D009765), weight loss (MESH:D015431), metabolic dysregulation (MESH:D021081), metabolic dysfunction (MESH:D008659), overnutrition (MESH:D044343), impaired (MESH:D060825), deficiencies (MESH:D007153), growth retardation (MESH:D006130), gut inflammation (MESH:D007249), immune dysfunction (MESH:D007154), Dysbiosis (MESH:D064806), PEM (MESH:D011502), underweight (MESH:D013851), psychiatric disorders (MESH:D001523)
- **Chemicals:** Methanol (MESH:D000432), PBS (MESH:D007854), Aspirin (MESH:D001241), Na2CO3 (MESH:C005686), Iron(II) sulfate heptahydrate (MESH:C020748), acids (MESH:D000143), amino acids (MESH:D000596), 2,4,6-tri(2-pyridyl)-s-triazine (MESH:C002849), CFE (-), phosphate (MESH:D010710), TCA (MESH:D014238), water (MESH:D014867), SCFAs (MESH:D005232), FeCl3 (MESH:C024555), K2S2O8 (MESH:C009007), tyrosine (MESH:D014443), agar (MESH:D000362), 2,2'-azino-bis(3-ethyl benzo thiazoline-6-sulfonic acid (MESH:C002502), acetate (MESH:D000085), 2,2-diphenyl-1-picrylhydrazyl (MESH:C004931), metal (MESH:D008670), Ascorbic acid (MESH:D001205)
- **Species:** Leptospira sp. AB (species) [taxon 103236], Lactiplantibacillus plantarum (species) [taxon 1590], Lactococcus lactis (species) [taxon 1358], Lactococcus cremoris (species) [taxon 1359], Proteus mirabilis (species) [taxon 584], Lacticaseibacillus paracasei (species) [taxon 1597], Bos taurus (bovine, species) [taxon 9913], Ligilactobacillus agilis (species) [taxon 1601], Bifidobacterium (genus) [taxon 1678], Lacticaseibacillus rhamnosus (species) [taxon 47715], Anaerococcus prevotii (species) [taxon 33034], Limosilactobacillus fermentum (species) [taxon 1613], Bacillus licheniformis (species) [taxon 1402], Bacillus tequilensis (species) [taxon 227866], Homo sapiens (human, species) [taxon 9606], Bacillus subtilis (species) [taxon 1423], Lacticaseibacillus casei (species) [taxon 1582]
- **Cell lines:** CI — Homo sapiens (Human), Chronic lymphocytic leukemia, Transformed cell line (CVCL_Y548)

## Full text

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## Figures

4 figures with captions in the complete paper: https://tomesphere.com/paper/PMC13012502/full.md

## References

42 references — full list in the complete paper: https://tomesphere.com/paper/PMC13012502/full.md

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Source: https://tomesphere.com/paper/PMC13012502