Genetic and biotechnological characterization of folate-producing probiotics isolated from local dairy products

Abstract Background Folate is essential for Deoxyribonucleic acid (DNA) synthesis, methylation and repair, with deficiencies linked to neural tube defects, Alzheimer’s, and cancer. Certain lactic acid bacteria (LAB), particularly in fermented dairy products, are natural folate producers. However, ge...

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Main Authors: Rahma Ali Hassan Abubakr, Mohammed Hassan Abdel A’al, Gihan Mohamed El Moghazy, Ashraf Bakry Abd Elrazik, Samir Abdel Aziz Ibrahim
Format: Article
Language:English
Published: SpringerOpen 2025-06-01
Series:Beni-Suef University Journal of Basic and Applied Sciences
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Online Access:https://doi.org/10.1186/s43088-025-00650-w
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author Rahma Ali Hassan Abubakr
Mohammed Hassan Abdel A’al
Gihan Mohamed El Moghazy
Ashraf Bakry Abd Elrazik
Samir Abdel Aziz Ibrahim
author_facet Rahma Ali Hassan Abubakr
Mohammed Hassan Abdel A’al
Gihan Mohamed El Moghazy
Ashraf Bakry Abd Elrazik
Samir Abdel Aziz Ibrahim
author_sort Rahma Ali Hassan Abubakr
collection DOAJ
description Abstract Background Folate is essential for Deoxyribonucleic acid (DNA) synthesis, methylation and repair, with deficiencies linked to neural tube defects, Alzheimer’s, and cancer. Certain lactic acid bacteria (LAB), particularly in fermented dairy products, are natural folate producers. However, genetic insights into high-yielding strains remain limited. This study isolates indigenous lactic acid bacteria (LAB) and provides the first genetic mapping of folate biosynthesis in Streptococcus thermophilus (S. thermophilus). Methods Probiotic bacteria were isolated from locally sourced milk and yogurt samples and were subjected to morphological, biochemical, and molecular identification. The isolates were screened for folate production levels. Molecular studies and “in-silico” analysis were carried out to investigate the genetic basis of folate biosynthesis, particularly in S. thermophilus. Results Two folate-producing probiotic strains, Enterococcus faecium (E. faecium) and S. thermophilus, were isolated and characterized. Quantitative analysis revealed folate production levels of 2.2–8.3 µg/L and 156–162 µg/L, respectively, demonstrating significantly higher synthesis by S. thermophilus. In this strain, seven folate biosynthesis genes and one metabolism gene were identified and mapped. Five of the biosynthetic genes were found to be clustered: folC2 (encoding dihydrofolate synthase), folE (encoding GTP cyclohydrolase I), folP (encoding dihydropteroate synthase), folB (encoding dihydroneopterin aldolase, and folK (encoding 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase). The remaining genes were dispersed throughout the genome. A conserved Shine-Dalgarno sequence (AGGAG) was found 5 base-pair (bp) upstream of five genes. Conclusion This study elucidates the genetic basis of folate production in S. thermophilus, enabling future metabolic engineering for enhanced folate yields. The findings support strain selection for folate-enriched functional foods.
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spelling doaj-art-4e55d12988044fb08ff2af1f44b2318f2025-08-20T02:06:35ZengSpringerOpenBeni-Suef University Journal of Basic and Applied Sciences2314-85432025-06-0114111510.1186/s43088-025-00650-wGenetic and biotechnological characterization of folate-producing probiotics isolated from local dairy productsRahma Ali Hassan Abubakr0Mohammed Hassan Abdel A’al1Gihan Mohamed El Moghazy2Ashraf Bakry Abd Elrazik3Samir Abdel Aziz Ibrahim4Ainshams UniversityThe Regional Center for Food and Feed (RCFF), Agriculture Research Center (ARC)The Regional Center for Food and Feed (RCFF), Agriculture Research Center (ARC)Ainshams UniversityAinshams UniversityAbstract Background Folate is essential for Deoxyribonucleic acid (DNA) synthesis, methylation and repair, with deficiencies linked to neural tube defects, Alzheimer’s, and cancer. Certain lactic acid bacteria (LAB), particularly in fermented dairy products, are natural folate producers. However, genetic insights into high-yielding strains remain limited. This study isolates indigenous lactic acid bacteria (LAB) and provides the first genetic mapping of folate biosynthesis in Streptococcus thermophilus (S. thermophilus). Methods Probiotic bacteria were isolated from locally sourced milk and yogurt samples and were subjected to morphological, biochemical, and molecular identification. The isolates were screened for folate production levels. Molecular studies and “in-silico” analysis were carried out to investigate the genetic basis of folate biosynthesis, particularly in S. thermophilus. Results Two folate-producing probiotic strains, Enterococcus faecium (E. faecium) and S. thermophilus, were isolated and characterized. Quantitative analysis revealed folate production levels of 2.2–8.3 µg/L and 156–162 µg/L, respectively, demonstrating significantly higher synthesis by S. thermophilus. In this strain, seven folate biosynthesis genes and one metabolism gene were identified and mapped. Five of the biosynthetic genes were found to be clustered: folC2 (encoding dihydrofolate synthase), folE (encoding GTP cyclohydrolase I), folP (encoding dihydropteroate synthase), folB (encoding dihydroneopterin aldolase, and folK (encoding 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase). The remaining genes were dispersed throughout the genome. A conserved Shine-Dalgarno sequence (AGGAG) was found 5 base-pair (bp) upstream of five genes. Conclusion This study elucidates the genetic basis of folate production in S. thermophilus, enabling future metabolic engineering for enhanced folate yields. The findings support strain selection for folate-enriched functional foods.https://doi.org/10.1186/s43088-025-00650-wProbioticsStreptococcus thermophilesEnterococcus faeciumFolateFolic acidBiosynthetic pathway
spellingShingle Rahma Ali Hassan Abubakr
Mohammed Hassan Abdel A’al
Gihan Mohamed El Moghazy
Ashraf Bakry Abd Elrazik
Samir Abdel Aziz Ibrahim
Genetic and biotechnological characterization of folate-producing probiotics isolated from local dairy products
Beni-Suef University Journal of Basic and Applied Sciences
Probiotics
Streptococcus thermophiles
Enterococcus faecium
Folate
Folic acid
Biosynthetic pathway
title Genetic and biotechnological characterization of folate-producing probiotics isolated from local dairy products
title_full Genetic and biotechnological characterization of folate-producing probiotics isolated from local dairy products
title_fullStr Genetic and biotechnological characterization of folate-producing probiotics isolated from local dairy products
title_full_unstemmed Genetic and biotechnological characterization of folate-producing probiotics isolated from local dairy products
title_short Genetic and biotechnological characterization of folate-producing probiotics isolated from local dairy products
title_sort genetic and biotechnological characterization of folate producing probiotics isolated from local dairy products
topic Probiotics
Streptococcus thermophiles
Enterococcus faecium
Folate
Folic acid
Biosynthetic pathway
url https://doi.org/10.1186/s43088-025-00650-w
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