Guinane Caitriona M, Piper Clare, Draper Lorraine A, O'Connor Paula M, Hill Colin, Ross R Paul, Cotter Paul D
Teagasc Food Research Centre, Moorepark, Fermoy, County Cork, Ireland.
APC Microbiome Institute, University College Cork, Cork, Ireland School of Microbiology, University College Cork, Cork, Ireland.
Appl Environ Microbiol. 2015 Nov;81(22):7851-9. doi: 10.1128/AEM.02339-15. Epub 2015 Sep 4.
Bacteriocin production is regarded as a desirable probiotic trait that aids in colonization and persistence in the gastrointestinal tract (GIT). Strains of Lactobacillus salivarius, a species associated with the GIT, are regarded as promising probiotic candidates and have a number of associated bacteriocins documented to date. These include multiple class IIb bacteriocins (salivaricin T, salivaricin P, and ABP-118) and the class IId bacteriocin bactofencin A, which show activity against medically important pathogens. However, the production of a bacteriocin in laboratory media does not ensure production under stressful environmental conditions, such as those encountered within the GIT. To allow this issue to be addressed, the promoter regions located upstream of the structural genes encoding the L. salivarius bacteriocins mentioned above were fused to a number of reporter proteins (green fluorescent protein [GFP], red fluorescent protein [RFP], and luciferase [Lux]). Of these, only transcriptional fusions to GFP generated signals of sufficient strength to enable the study of promoter activity in L. salivarius. While analysis of the class IIb bacteriocin promoter regions indicated relatively weak GFP expression, assessment of the promoter of the antistaphylococcal bacteriocin bactofencin A revealed a strong promoter that is most active in the absence of the antimicrobial peptide and is positively induced in the presence of mild environmental stresses, including simulated gastric fluid. Taken together, these data provide information on factors that influence bacteriocin production, which will assist in the development of strategies to optimize in vivo and in vitro production of these antimicrobials.
细菌素的产生被视为一种理想的益生菌特性,有助于在胃肠道(GIT)中定殖和持续存在。唾液乳杆菌是一种与胃肠道相关的细菌,其菌株被认为是有前景的益生菌候选者,并且迄今为止已记录了许多相关的细菌素。这些包括多种IIb类细菌素(唾液乳杆菌素T、唾液乳杆菌素P和ABP-118)以及IId类细菌素杆菌防御素A,它们对医学上重要的病原体具有活性。然而,在实验室培养基中产生细菌素并不能确保在应激环境条件下(如在胃肠道中遇到的条件)也能产生。为了解决这个问题,将上述编码唾液乳杆菌细菌素的结构基因上游的启动子区域与多种报告蛋白(绿色荧光蛋白[GFP]、红色荧光蛋白[RFP]和荧光素酶[Lux])融合。其中,只有与GFP的转录融合产生了足够强度的信号,以便能够研究唾液乳杆菌中的启动子活性。虽然对IIb类细菌素启动子区域的分析表明GFP表达相对较弱,但对抗葡萄球菌细菌素杆菌防御素A的启动子的评估揭示了一个强启动子,该启动子在没有抗菌肽的情况下最活跃,并且在存在包括模拟胃液在内的轻度环境应激时被正向诱导。综上所述,这些数据提供了有关影响细菌素产生的因素的信息,这将有助于制定优化这些抗菌剂体内和体外生产的策略。