Department of Biochemistry, Physiology and Microbiology, Ghent University, Belgium.
J Appl Microbiol. 2009 Apr;106(4):1081-92. doi: 10.1111/j.1365-2672.2008.04094.x. Epub 2009 Jan 30.
To investigate the circulation of predominant sourdough lactic acid bacteria (LAB) species in the production environment of two Belgian artisan sourdough bakeries.
Isolates were collected from sourdoughs, flour, hands of the baker and air in the bakery setting and taxonomically characterized using repetitive element sequence-based PCR fingerprinting, pheS and/or 16S rRNA gene sequencing and amplified fragment length polymorphism (AFLP) analysis. In parallel, PCR-DGGE (denaturing gradient gel electrophoresis) analysis of V3-16S rDNA amplicons was applied to visualize the predominant bacterial population in the sourdoughs and the corresponding bakery environment (flour, hands of the baker, air and bakery equipment). Both approaches revealed that sourdoughs produced at D01 and D10 were mainly dominated by Lactobacillus spicheri and L. plantarum and by L. sanfranciscensis, respectively, and that these LAB species also circulated in the corresponding bakery environment. Furthermore, AFLP fingerprinting demonstrated that sourdough and bakery environment isolates of these species were genetically indistinguishable. For more sensitive source-tracking, SYBR Green-based real-time PCR assays were developed using species-specific primers targeting the pheS gene of L. plantarum and L. sanfranciscensis, detected in air samples from D01 and D10, respectively.
The results obtained in this study indicate that specific strains of LAB persist in artisan doughs over years and circulate in the bakery environment. Furthermore, the importance of air as a potential carrier of LAB in artisan bakery environments was demonstrated.
PheS-based real-time PCR can be used to detect, quantify and/or monitor specific LAB species (e.g. starter cultures) in sourdough and bakery environment samples.
调查两家比利时传统酸面团面包店生产环境中主要的酸面团乳酸菌(LAB)种类的循环情况。
从酸面团、面粉、面包师的手和面包店内空气中采集分离物,并使用重复元件序列 PCR 指纹分析、pheS 和/或 16S rRNA 基因测序和扩增片段长度多态性(AFLP)分析对其进行分类鉴定。同时,应用 PCR-DGGE(变性梯度凝胶电泳)分析 V3-16S rDNA 扩增子,以可视化酸面团和相应面包店环境(面粉、面包师的手、空气和面包店设备)中的主要细菌种群。这两种方法都表明,D01 和 D10 生产的酸面团主要由 Lactobacillus spicheri 和 L. plantarum 以及 L. sanfranciscensis 主导,这些 LAB 种类也在相应的面包店环境中循环。此外,AFLP 指纹分析表明,这些种类的酸面团和面包店环境分离物在遗传上是无法区分的。为了更灵敏地进行溯源,针对 L. plantarum 和 L. sanfranciscensis 的 pheS 基因,分别开发了 SYBR Green 基于实时 PCR 检测法,检测到 D01 和 D10 的空气样本中存在这两种基因。
本研究结果表明,特定的 LAB 菌株在多年来在传统面团中持续存在,并在面包店内环境中循环。此外,空气作为传统面包店内环境中 LAB 的潜在载体的重要性得到了证明。
pheS 基于实时 PCR 可用于检测、定量和/或监测酸面团和面包店内环境样品中的特定 LAB 种类(例如,发酵剂)。