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Using the Overlay Assay to Qualitatively Measure Bacterial Production of and Sensitivity to Pneumococcal Bacteriocins
Published on: September 30, 2014
Bifidobacterium longum subsp. infantis as widespread bacteriocin gene clusters carrier stands out among the
Di Yu1,2, Zhangming Pei1,2, Yutao Chen1,2
1State Key Laboratory of Food Science and Resources, Jiangnan University , Wuxi, Jiangsu, China.
Insights
Bacteriocin gene clusters were identified in Bifidobacterium, particularly in B. infantis, revealing their role in infant gut colonization. This study highlights Bifidobacterium
Area of Science:
- Microbiology and Genomics
- Bacteriocin Discovery
- Infant Gut Microbiota
Background:
- Bifidobacterium is a dominant genus in healthy breast-fed infants, crucial for gut microbiota balance and infant development.
- Bacteriocin production in Bifidobacterium has been historically underestimated due to limited isolation and identification.
- Understanding the genetic basis for Bifidobacterium's adaptation to the infant gut niche is essential.
Purpose of the Study:
- To comprehensively re-evaluate the distribution and potential of bacteriocin production in Bifidobacterium species.
- To identify novel bacteriocin gene clusters (BGCs) in Bifidobacterium genomes using a silicon-based search strategy.
- To investigate the role of bacteriocins in the competitive colonization of the infant intestinal tract by Bifidobacterium.
Main Methods:
- Bioinformatic analysis of 759 Bifidobacterium genomes from 9 species to mine for putative bacteriocin gene clusters (BGCs).
- Utilized a silicon-based search strategy to identify 354 BGCs, many previously unreported.
- Performed antimicrobial activity assays and transcription analysis to confirm bacteriocin biosynthesis potential.
Main Results:
- Discovered 354 putative bacteriocin gene clusters (BGCs) across 759 Bifidobacterium strains.
- Bifidobacterium longum subsp. infantis strains were found to carry numerous BGCs, including lanthipeptides, lasso peptides, thiopeptides, and class IId bacteriocins.
- Antimicrobial activity and transcription analysis confirmed the biosynthetic potential of identified bacteriocins.
Conclusions:
- This study reveals a significant underestimation of bacteriocin production capabilities in Bifidobacterium, especially B. infantis.
- Bacteriocin genes provide a competitive advantage for Bifidobacterium species, aiding colonization in the infant intestinal tract.
- The findings offer new insights into the genetic mechanisms underlying Bifidobacterium's success in the early-life gut environment.
Abstract:
Bifidobacterium is the dominant genus, particularly in the intestinal tract niche of healthy breast-fed infants, and many of these strains have been proven to elicit positive effects on infant development. In addition to its effective antimicrobial activity against detrimental microorganisms, it helps to improve the intestinal microbiota balance. The isolation and identification of bacteriocins from Bifidobacterium have been limited since the mid-1980s, leading to an underestimation of its ability for bacteriocin production. Here, we employed a silicon-based search strategy to mine 354 putative bacteriocin gene clusters (BGCs), most of which have never been reported, from the genomes of 759 Bifidobacterium strains distributed across 9 species. Consistent with previous reports, most Bifidobacterium strains did not carry or carry only a single BGC; however, Bifidobacterium longum subsp. infantis, in contrast to other Bifidobacterium species, carried numerous BGCs, including lanthipeptides, lasso peptides, thiopeptides, and class IId bacteriocins. The antimicrobial activity of the crude bacteriocins and transcription analysis confirmed its potential for bacteriocin biosynthesis. Additionally, we investigated the association of bacteriocins with the phylogenetic positions of their homologs from other genera and niches. In conclusion, this study re-examines a few Bifidobacterium species traditionally regarded as a poor source of bacteriocins. These bacteriocin genes impart a competitive advantage to Bifidobacterium in colonizing the infant intestinal tract. IMPORTANCE Development of the human gut microbiota commences from birth, with bifidobacteria being among the first colonizers of the newborn intestinal tract and dominating it for a considerable period. To date, the genetic basis for the successful adaptation of bifidobacteria to this particular niche remains unclear since studies have mainly focused on glycoside hydrolase and adhesion-related genes. Bacteriocins are competitive factors that help producers maintain colonization advantages without destroying the niche balance; however, they have rarely been reported in Bifidobacterium. The advancement in sequencing methods and bacteriocin databases enables the use of a silicon-based search strategy for the comprehensive and rapid re-evaluation of the bacteriocin distribution of Bifidobacterium. Our study revealed that B. infantis carries abundant bacteriocin biosynthetic gene clusters for the first time, presenting new evidence regarding the competitive interactions of Bifidobacterium in the infant intestinal tract.
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