Physicochemical, Biochemical, Microbiological, and Sensory Characterization of a Fermented Lima Bean (Phaseolus lunatus) Probiotic Beverage During Refrigerated Storage

Hassina Bilek; Ana Lúcia  Oliveira; Manuela  Amorim; Ana Raquel  Madureira; Farid Zaid; Maria Manuela  Pintado

doi:10.51745/najfnr.10.21.178-191

Physicochemical, Biochemical, Microbiological, and Sensory Characterization of a Fermented Lima Bean (Phaseolus lunatus) Probiotic Beverage During Refrigerated Storage

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Food Chemistry, Engineering, Processing and Packaging Food Microbiology, Safety and Toxicology Functional and Novel Foods

DOI https://doi.org/10.51745/najfnr.10.21.178-191

Issue Vol. 10 No. 21 (2026): January - June, 2026

Submitted 2025-05-12

Accepted 2026-06-13

Published 2026-06-26

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Background: Growing public health concerns regarding the consumption of conventional dairy products, coupled with the increasing prevalence of plant-based and vegan dietary preferences, have stimulated considerable scientific interest in the development of innovative fermented legume-based beverages. The lima bean (Phaseolus lunatus) represents a particularly promising raw material in this context, due to its high protein content, the presence of prebiotic α-galactosides, and its notable endogenous antioxidant compounds.

Aims: This study aimed to evaluate the probiotic viability, physicochemical and biochemical properties, sensory characteristics, and antioxidant potential of fermented lima bean-based beverages subjected to refrigerated storage at 4°C for 28 days.

Material and Methods: Four lima bean-based beverages (Lbs) were formulated by incorporating a 1% inoculum of either the single-strain probiotic Lactobacillus acidophilus La5 (LbLa5), Bifidobacterium animalis subsp. lactis Bb12 (LbBb12), a co-culture of both strains (Lb coculture), or no bacterial inoculum, serving as a non-fermented control beverage.

Results: The probiotic Lbs exhibited a high crude protein content (3.15% – 3.34%) comparable to that of soy-based and bovine milk, and were characterized by a substantial carbohydrate fraction (2.08% – 2.32%). Viable counts of L. acidophilus La5 and B. lactis Bb12 were maintained at 6 – 7 log CFU/mL throughout the shelf-life period, with B. lactis Bb12 demonstrating enhanced survival under co-culture conditions. High-performance liquid chromatography (HPLC) analysis revealed significant reduction in the prebiotic oligosaccharide raffinose throughout storage with final concentrations ranging from 2.17 to 3.35 g/L. Probiotic metabolic activity induced a progressive decline in pH (4.00 – 4.30) accompanied by an increase in titratable acidity (0.70 – 0.89%). Chromatographic profiles demonstrated that both selected strains were proteolytically active during cold storage, resulting in a reduction of free amino acid content and the generation of bioactive low-molecular-weight peptides (≤ 10 kDa). Fermentation promoted an increase in total phenolic compound (TPC) content over the first seven days, whilst DPPH radical scavenging activity increased over the initial three weeks of storage. The co-culture treatment yielded the highest TPC content (1.71 – 2.03 mg GAE/mL) and DPPH radical scavenging activity (0.86 – 1.52 mg TE/mL) throughout the entire storage period. Sensory flavor attributes and overall acceptability were enhanced across all fermented beverage formulations relative to the non-fermented control.

Conclusions: The incorporation of L. acidophilus La5 and B. lactis Bb12 into lima bean-based beverages yields a nutritionally dense, functionally enriched, and sensorially acceptable plant-based alternative well-suited to vegan and health-conscious dietary regimens. These findings support the valorization of Phaseolus lunatus as a viable substrate for the development of probiotic fermented beverages.

Keywords

Lima Bean-Based Beverage Probiotics Raffinose Organic Acids Free Amino Acids Bioactive Peptides Antioxidant Capacity

Hassina Bilek

Département des Sciences Alimentaires, Faculté des Sciences de la Nature et de la Vie, Université de Bejaia, Route Targa Ouzemour, Bejaia 06000, Algeria
Ana Lúcia Oliveira

Universidade Católica Portuguesa
Manuela Amorim

Universidade Católica Portuguesa
Ana Raquel Madureira

Universidade Católica Portuguesa
Farid Zaid

Département des Sciences Alimentaires, Faculté des Sciences de la Nature et de la Vie, Université de Bejaia, Route Targa Ouzemour, Bejaia 06000, Algeria
Maria Manuela Pintado

Universidade Católica Portuguesa

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Bilek, H., Oliveira, A. L. ., Amorim, M. ., Madureira, A. R. ., Zaid, F., & Pintado, M. M. . (2026). Physicochemical, Biochemical, Microbiological, and Sensory Characterization of a Fermented Lima Bean (Phaseolus lunatus) Probiotic Beverage During Refrigerated Storage. The North African Journal of Food and Nutrition Research, 10(21), 178-191. https://doi.org/10.51745/najfnr.10.21.178-191

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Physicochemical, Biochemical, Microbiological, and Sensory Characterization of a Fermented Lima Bean (Phaseolus lunatus) Probiotic Beverage During Refrigerated Storage

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