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Abstract <p><i>Lactica</i><i>seibacillus paracasei</i> INIA P708 was isolated from breast-fed infant feces. It stood out by its stable ropy phenotype, which could indicate exopolysaccharide (EPS) production and antimicrobial properties. Bearing in mind its potential use in the food chain, the objective of this work was to study <i>L. paracasei</i> INIA P708 potential probiotic traits, and technological and safety characteristics, in comparison to the commercial probiotic <i>L. rhamnosus</i> GG strain. A full genomic analysis was also performed to comply with EFSA requirement for microorganisms intentionally added to the food chain. EPS production was tested by confocal scanning laser microscopy (CSLM), labelling bacterial cultures with fluorescent lectin conjugates. As probiotic traits, <i>in vitro</i> survival under major gastrointestinal conditions, biofilm formation, and antimicrobial activity was tested. As technological properties, survival as lyophilized cultures, growth in milk, proteolytic and acidifying properties, salt tolerance, and growth with prebiotic compounds were studied. As safety traits, antibiotic resistance, biogenic amine production, and mucine degradation were also tested. The complete, closed, INIA P708 genome sequence was determined by combining long and short reads. CSLM confirmed EPS production by <i>L. paracasei</i> INIA P708, and their attachment to the cell surface. INIA P708 showed good survival rates after lysozyme exposure (98%) and after simulated major GIT conditions (97%). However, under the tested conditions, and opposite to what we observed for GG, INIA P708 was not able to form biofilm. Both strains showed a clear inhibition halo against <i>Listeria monocytogenes</i> and a diffuse halo against <i>Klebsiella oxytoca</i>. The lyophilization process lowered levels by 0.2–0.3 log units for both strains, without changes after 21&#xa0;days of refrigerated storage. Salt had a significant effect on the growth of both strains, with INIA P708 being more sensitive to salt than GG. Regarding safety traits, both strains were susceptible to chloramphenicol, erythromycin, rifampicin, and tetracycline and resistant to kanamycin and vancomycin. They were neither able to produce biogenic amines from histidine or tyrosine nor to degrade mucine. The closed INIA P708 genome is composed by a chromosome (3.08 Mpb) and a plasmid (66.8 Kpb), with a GC content of 46.3%. Detailed genome analysis confirmed <i>L. paracasei</i> INIA P708 identification and safety, supported its use as an industrial strain, and revealed new potential probiotic traits in this strain.</p> Key points <p>• <i>Lacticaseibacillus paracasei INIA P708 is a human isolate with high potential.</i></p> <p>• <i>It might be used in health-promoting and/or in technological applications.</i></p> <p>• <i>Full genome resolution confirms it could be safely used as an industrial strain.</i></p>

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Lacticaseibacillus paracasei INIA P708: a safe human isolate with probiotic and technological potential

  • 期刊论文

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  • 发表日期：

    2026年8月4日

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  • Eduardo D. Lozano-Escobar,
  • Eva Rodríguez-Mínguez,
  • Karel H. M. van Wely,
  • Ángela Peirotén,
  • Modesto Redrejo-Rodríguez,
  • Antonia Picon

摘要

Abstract

Lacticaseibacillus paracasei INIA P708 was isolated from breast-fed infant feces. It stood out by its stable ropy phenotype, which could indicate exopolysaccharide (EPS) production and antimicrobial properties. Bearing in mind its potential use in the food chain, the objective of this work was to study L. paracasei INIA P708 potential probiotic traits, and technological and safety characteristics, in comparison to the commercial probiotic L. rhamnosus GG strain. A full genomic analysis was also performed to comply with EFSA requirement for microorganisms intentionally added to the food chain. EPS production was tested by confocal scanning laser microscopy (CSLM), labelling bacterial cultures with fluorescent lectin conjugates. As probiotic traits, in vitro survival under major gastrointestinal conditions, biofilm formation, and antimicrobial activity was tested. As technological properties, survival as lyophilized cultures, growth in milk, proteolytic and acidifying properties, salt tolerance, and growth with prebiotic compounds were studied. As safety traits, antibiotic resistance, biogenic amine production, and mucine degradation were also tested. The complete, closed, INIA P708 genome sequence was determined by combining long and short reads. CSLM confirmed EPS production by L. paracasei INIA P708, and their attachment to the cell surface. INIA P708 showed good survival rates after lysozyme exposure (98%) and after simulated major GIT conditions (97%). However, under the tested conditions, and opposite to what we observed for GG, INIA P708 was not able to form biofilm. Both strains showed a clear inhibition halo against Listeria monocytogenes and a diffuse halo against Klebsiella oxytoca. The lyophilization process lowered levels by 0.2–0.3 log units for both strains, without changes after 21 days of refrigerated storage. Salt had a significant effect on the growth of both strains, with INIA P708 being more sensitive to salt than GG. Regarding safety traits, both strains were susceptible to chloramphenicol, erythromycin, rifampicin, and tetracycline and resistant to kanamycin and vancomycin. They were neither able to produce biogenic amines from histidine or tyrosine nor to degrade mucine. The closed INIA P708 genome is composed by a chromosome (3.08 Mpb) and a plasmid (66.8 Kpb), with a GC content of 46.3%. Detailed genome analysis confirmed L. paracasei INIA P708 identification and safety, supported its use as an industrial strain, and revealed new potential probiotic traits in this strain.

Key points

• Lacticaseibacillus paracasei INIA P708 is a human isolate with high potential.

• It might be used in health-promoting and/or in technological applications.

• Full genome resolution confirms it could be safely used as an industrial strain.