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<p>This study evaluated the effects of fermentation time (24, 48, and 72&#xa0;h), and post-fermentation heat treatment (PFHT) on yield, physicochemical properties, antioxidant activity, and antimicrobial activity of postbiotics produced from the co-fermentation of <i>Lactobacillus rhamnosus</i> GG, <i>Lactobacillus plantarum</i>,<i> Bifidobacterium longum</i>, and <i>Bifidobacterium breve</i>. Cross-streak assays confirmed strain compatibility under anaerobic conditions. Net postbiotic yield (dry mass corrected against an uninoculated control) was significantly influenced by fermentation time and PFHT (<i>P</i> &lt; 0.05), with the highest yield observed at 48&#xa0;h under PFHT (70.97 ± 0.36&#xa0;g/L). Fermentation was characterized by progressive acidification (pH 6.37–3.81) and biomass accumulation (OD₆₀₀: 0.15–1.66). The solubility of freeze-dried postbiotics was 95–98% and was not significantly affected by fermentation time or PFHT (<i>P</i> &gt; 0.05). Antioxidant activity increased with fermentation time and was significantly higher in PFHT samples, with DPPH (62.09–83.43%) and ABTS⁺ (79.82–95.62%) scavenging activities, reaching a plateau at 48&#xa0;h. Antimicrobial activity was greater in PFHT samples, with the largest inhibition zones of 14.33 ± 0.58&#xa0;mm were observed at 48&#xa0;h. The correlation and principal component analyses indicated associations between acidification and antioxidant activity, although causality cannot be inferred. Within the limitations of this study, including a single heat treatment condition, absence of minimum inhibitory concentration data, lack of metabolomic characterization, and exclusive in vitro evaluation, the findings suggest that 48&#xa0;h fermentation combined with PFHT provides a favorable balance of postbiotic yield and bioactivity. Further studies are required to support optimization and application.</p>

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Effect of Fermentation Time and Heat Treatment on Yield and In Vitro Antioxidant and Antimicrobial Activity of Postbiotics from Mixed Bifidobacterium and Lactobacillus Co-Fermentations

  • 期刊论文

  • 发表日期：

    2026年6月3日

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  • Zerihun Asefa,
  • Abera Belay,
  • Teferi Damto,
  • Admasu Addi,
  • Endale Amare,
  • Feriehiwote Weldeyohanis Gebremariam,
  • Ibrahim Ahmed Sherif,
  • Henock Woldemichael Woldemariam

摘要

This study evaluated the effects of fermentation time (24, 48, and 72 h), and post-fermentation heat treatment (PFHT) on yield, physicochemical properties, antioxidant activity, and antimicrobial activity of postbiotics produced from the co-fermentation of Lactobacillus rhamnosus GG, Lactobacillus plantarum, Bifidobacterium longum, and Bifidobacterium breve. Cross-streak assays confirmed strain compatibility under anaerobic conditions. Net postbiotic yield (dry mass corrected against an uninoculated control) was significantly influenced by fermentation time and PFHT (P < 0.05), with the highest yield observed at 48 h under PFHT (70.97 ± 0.36 g/L). Fermentation was characterized by progressive acidification (pH 6.37–3.81) and biomass accumulation (OD₆₀₀: 0.15–1.66). The solubility of freeze-dried postbiotics was 95–98% and was not significantly affected by fermentation time or PFHT (P > 0.05). Antioxidant activity increased with fermentation time and was significantly higher in PFHT samples, with DPPH (62.09–83.43%) and ABTS⁺ (79.82–95.62%) scavenging activities, reaching a plateau at 48 h. Antimicrobial activity was greater in PFHT samples, with the largest inhibition zones of 14.33 ± 0.58 mm were observed at 48 h. The correlation and principal component analyses indicated associations between acidification and antioxidant activity, although causality cannot be inferred. Within the limitations of this study, including a single heat treatment condition, absence of minimum inhibitory concentration data, lack of metabolomic characterization, and exclusive in vitro evaluation, the findings suggest that 48 h fermentation combined with PFHT provides a favorable balance of postbiotic yield and bioactivity. Further studies are required to support optimization and application.