Evaluation of inter‐individual differences in gut bacterial isoflavone bioactivation in humans by PCR‐based targeting of genes involved in equol formation
Aim To identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR‐based approach. Methods and Results In a pilot study including 17 human subjects, equol formation was determined in faecal slurries. In parallel, faecal DNA was amplified by PCR usin...
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| Published in: | Journal of applied microbiology Vol. 124; no. 1; pp. 220 - 231 |
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| Main Authors: | , |
| Format: | Journal Article |
| Language: | English |
| Published: |
England
Oxford University Press
01.01.2018
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| ISSN: | 1364-5072, 1365-2672, 1365-2672 |
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| Abstract | Aim
To identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR‐based approach.
Methods and Results
In a pilot study including 17 human subjects, equol formation was determined in faecal slurries. In parallel, faecal DNA was amplified by PCR using degenerate primers that target highly conserved regions of dihydrodaidzein reductase and tetrahydrodaidzein reductase genes. PCR products of the expected size were observed for six of the eight subjects identified as equol producers. Analysis of clone libraries revealed the amplification of sequences exclusively related to Adlercreutzia equolifaciens in four of the subjects tested positive for equol formation, whereas in three of the equol producers, only sequences related to Slackia isoflavoniconvertens were observed. No amplicons were obtained for one equol‐forming subject, thus suggesting the presence of nontargeted alternative genes. Amplicons were only sporadically observed in the nonequol producers.
Conclusion
The majority of human subjects who produced equol were also detected with the developed PCR‐based approach.
Significance and Impact of the Study
The obtained results shed light on the distribution and the diversity of known equol‐forming bacterial species in the study group and indicate the presence of as yet unknown equol‐forming bacteria. |
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| AbstractList | To identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR-based approach.AIMTo identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR-based approach.In a pilot study including 17 human subjects, equol formation was determined in faecal slurries. In parallel, faecal DNA was amplified by PCR using degenerate primers that target highly conserved regions of dihydrodaidzein reductase and tetrahydrodaidzein reductase genes. PCR products of the expected size were observed for six of the eight subjects identified as equol producers. Analysis of clone libraries revealed the amplification of sequences exclusively related to Adlercreutzia equolifaciens in four of the subjects tested positive for equol formation, whereas in three of the equol producers, only sequences related to Slackia isoflavoniconvertens were observed. No amplicons were obtained for one equol-forming subject, thus suggesting the presence of nontargeted alternative genes. Amplicons were only sporadically observed in the nonequol producers.METHODS AND RESULTSIn a pilot study including 17 human subjects, equol formation was determined in faecal slurries. In parallel, faecal DNA was amplified by PCR using degenerate primers that target highly conserved regions of dihydrodaidzein reductase and tetrahydrodaidzein reductase genes. PCR products of the expected size were observed for six of the eight subjects identified as equol producers. Analysis of clone libraries revealed the amplification of sequences exclusively related to Adlercreutzia equolifaciens in four of the subjects tested positive for equol formation, whereas in three of the equol producers, only sequences related to Slackia isoflavoniconvertens were observed. No amplicons were obtained for one equol-forming subject, thus suggesting the presence of nontargeted alternative genes. Amplicons were only sporadically observed in the nonequol producers.The majority of human subjects who produced equol were also detected with the developed PCR-based approach.CONCLUSIONThe majority of human subjects who produced equol were also detected with the developed PCR-based approach.The obtained results shed light on the distribution and the diversity of known equol-forming bacterial species in the study group and indicate the presence of as yet unknown equol-forming bacteria.SIGNIFICANCE AND IMPACT OF THE STUDYThe obtained results shed light on the distribution and the diversity of known equol-forming bacterial species in the study group and indicate the presence of as yet unknown equol-forming bacteria. To identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR-based approach. In a pilot study including 17 human subjects, equol formation was determined in faecal slurries. In parallel, faecal DNA was amplified by PCR using degenerate primers that target highly conserved regions of dihydrodaidzein reductase and tetrahydrodaidzein reductase genes. PCR products of the expected size were observed for six of the eight subjects identified as equol producers. Analysis of clone libraries revealed the amplification of sequences exclusively related to Adlercreutzia equolifaciens in four of the subjects tested positive for equol formation, whereas in three of the equol producers, only sequences related to Slackia isoflavoniconvertens were observed. No amplicons were obtained for one equol-forming subject, thus suggesting the presence of nontargeted alternative genes. Amplicons were only sporadically observed in the nonequol producers. The majority of human subjects who produced equol were also detected with the developed PCR-based approach. The obtained results shed light on the distribution and the diversity of known equol-forming bacterial species in the study group and indicate the presence of as yet unknown equol-forming bacteria. AIM: To identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR‐based approach. METHODS AND RESULTS: In a pilot study including 17 human subjects, equol formation was determined in faecal slurries. In parallel, faecal DNA was amplified by PCR using degenerate primers that target highly conserved regions of dihydrodaidzein reductase and tetrahydrodaidzein reductase genes. PCR products of the expected size were observed for six of the eight subjects identified as equol producers. Analysis of clone libraries revealed the amplification of sequences exclusively related to Adlercreutzia equolifaciens in four of the subjects tested positive for equol formation, whereas in three of the equol producers, only sequences related to Slackia isoflavoniconvertens were observed. No amplicons were obtained for one equol‐forming subject, thus suggesting the presence of nontargeted alternative genes. Amplicons were only sporadically observed in the nonequol producers. CONCLUSION: The majority of human subjects who produced equol were also detected with the developed PCR‐based approach. SIGNIFICANCE AND IMPACT OF THE STUDY: The obtained results shed light on the distribution and the diversity of known equol‐forming bacterial species in the study group and indicate the presence of as yet unknown equol‐forming bacteria. Aim To identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR‐based approach. Methods and Results In a pilot study including 17 human subjects, equol formation was determined in faecal slurries. In parallel, faecal DNA was amplified by PCR using degenerate primers that target highly conserved regions of dihydrodaidzein reductase and tetrahydrodaidzein reductase genes. PCR products of the expected size were observed for six of the eight subjects identified as equol producers. Analysis of clone libraries revealed the amplification of sequences exclusively related to Adlercreutzia equolifaciens in four of the subjects tested positive for equol formation, whereas in three of the equol producers, only sequences related to Slackia isoflavoniconvertens were observed. No amplicons were obtained for one equol‐forming subject, thus suggesting the presence of nontargeted alternative genes. Amplicons were only sporadically observed in the nonequol producers. Conclusion The majority of human subjects who produced equol were also detected with the developed PCR‐based approach. Significance and Impact of the Study The obtained results shed light on the distribution and the diversity of known equol‐forming bacterial species in the study group and indicate the presence of as yet unknown equol‐forming bacteria. AimTo identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR‐based approach.Methods and ResultsIn a pilot study including 17 human subjects, equol formation was determined in faecal slurries. In parallel, faecal DNA was amplified by PCR using degenerate primers that target highly conserved regions of dihydrodaidzein reductase and tetrahydrodaidzein reductase genes. PCR products of the expected size were observed for six of the eight subjects identified as equol producers. Analysis of clone libraries revealed the amplification of sequences exclusively related to Adlercreutzia equolifaciens in four of the subjects tested positive for equol formation, whereas in three of the equol producers, only sequences related to Slackia isoflavoniconvertens were observed. No amplicons were obtained for one equol‐forming subject, thus suggesting the presence of nontargeted alternative genes. Amplicons were only sporadically observed in the nonequol producers.ConclusionThe majority of human subjects who produced equol were also detected with the developed PCR‐based approach.Significance and Impact of the StudyThe obtained results shed light on the distribution and the diversity of known equol‐forming bacterial species in the study group and indicate the presence of as yet unknown equol‐forming bacteria. |
| Author | Braune, A. Blaut, M. |
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| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/29055162$$D View this record in MEDLINE/PubMed |
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| Keywords | daidzein equol isoflavone human intestinal microbiota dihydrodaidzein reductase tetrahydrodaidzein reductase |
| Language | English |
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To identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR‐based approach.
Methods and Results
In a... To identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR-based approach. In a pilot study including 17... AimTo identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR‐based approach.Methods and ResultsIn a... To identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR-based approach.AIMTo identify human subjects... AIM: To identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR‐based approach. METHODS AND RESULTS: In... |
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| SubjectTerms | Adult Bacteria Bacteria - genetics Bacteria - isolation & purification Bacteria - metabolism Bacterial Proteins - genetics Bacterial Proteins - metabolism Biodiversity Biological activity Daidzein Deoxyribonucleic acid digestive system dihydrodaidzein reductase DNA equol Equol - metabolism Feces - microbiology Female Forming Gastrointestinal Microbiome Gastrointestinal Tract - microbiology Genes human intestinal microbiota Human subjects Humans intestinal microorganisms Intestine isoflavone Isoflavones Isoflavones - metabolism Male metabolism Oxidoreductases - genetics Oxidoreductases - metabolism Pilot Projects Polymerase chain reaction Polymerase Chain Reaction - methods Primers Reductase Slackia Slurries tetrahydrodaidzein reductase |
| Title | Evaluation of inter‐individual differences in gut bacterial isoflavone bioactivation in humans by PCR‐based targeting of genes involved in equol formation |
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| Volume | 124 |
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