TI Food and Nutrition, Wageningen, The Netherlands.
PLoS One. 2012;7(7):e39053. doi: 10.1371/journal.pone.0039053. Epub 2012 Jul 3.
Lactic acid bacteria (LAB) are applied worldwide in the production of a variety of fermented food products. Additionally, specific Lactobacillus species are nowadays recognized for their health-promoting effects on the consumer. To optimally exert such beneficial effects, it is considered of great importance that these probiotic bacteria reach their target sites in the gut alive.
METHODOLOGY/PRINCIPAL FINDINGS: In the accompanying manuscript by Bron et al. the probiotic model organism Lactobacillus plantarum WCFS1 was cultured under different fermentation conditions, which was complemented by the determination of the corresponding molecular responses by full-genome transcriptome analyses. Here, the gastrointestinal (GI) survival of the cultures produced was assessed in an in vitro assay. Variations in fermentation conditions led to dramatic differences in GI-tract survival (up to 7-log) and high robustness could be associated with low salt and low pH during the fermentations. Moreover, random forest correlation analyses allowed the identification of specific transcripts associated with robustness. Subsequently, the corresponding genes were targeted by genetic engineering, aiming to enhance robustness, which could be achieved for 3 of the genes that negatively correlated with robustness and where deletion derivatives displayed enhanced survival compared to the parental strain. Specifically, a role in GI-tract survival could be confirmed for the lp_1669-encoded AraC-family transcription regulator, involved in capsular polysaccharide remodeling, the penicillin-binding protein Pbp2A involved in peptidoglycan biosynthesis, and the Na(+)/H(+) antiporter NapA3. Moreover, additional physiological analysis established a role for Pbp2A and NapA3 in bile salt and salt tolerance, respectively.
Transcriptome trait matching enabled the identification of biomarkers for bacterial (gut-)robustness, which is important for our molecular understanding of GI-tract survival and could facilitate the design of culture conditions aimed to enhance probiotic culture robustness.
乳酸菌(LAB)在全球范围内被应用于生产各种发酵食品。此外,特定的乳杆菌属物种因其对消费者的健康促进作用而受到认可。为了最佳发挥这些有益作用,益生菌细菌活着到达肠道的靶部位被认为非常重要。
方法/主要发现:在 Bron 等人的随附手稿中,对益生菌模式生物植物乳杆菌 WCFS1 进行了不同发酵条件下的培养,并通过全基因组转录组分析确定了相应的分子反应。在这里,通过体外试验评估了培养物在胃肠道(GI)中的存活情况。发酵条件的变化导致 GI 道存活(高达 7 个对数)的显著差异,并且高稳健性可与发酵过程中的低盐和低 pH 值相关联。此外,随机森林相关分析允许鉴定与稳健性相关的特定转录本。随后,通过遗传工程靶向相应的基因,旨在增强稳健性,这可以针对与稳健性呈负相关的 3 个基因实现,其中缺失衍生物与亲本菌株相比显示出增强的生存能力。具体而言,lp_1669 编码的 AraC 家族转录调节剂在参与荚膜多糖重塑的 GI 道生存中起作用,青霉素结合蛋白 Pbp2A 参与肽聚糖生物合成,Na(+) / H(+) 反向转运蛋白 NapA3。此外,额外的生理分析确立了 Pbp2A 和 NapA3 在胆盐和耐盐性中的作用。
转录组特征匹配使能够识别细菌(肠道)稳健性的生物标志物,这对我们理解 GI 道生存的分子机制非常重要,并可以促进旨在增强益生菌培养稳健性的培养条件的设计。