Duthoit Frédérique, Godon Jean-Jacques, Montel Marie-Christine
Laboratoire de Recherches Fromagères, INRA, 36 rue de Salers, 15000 Aurillac, France.
Appl Environ Microbiol. 2003 Jul;69(7):3840-8. doi: 10.1128/AEM.69.7.3840-3848.2003.
Microbial dynamics during processing and ripening of traditional cheeses such as registered designation of origin Salers cheese, an artisanal cheese produced in France, play an important role in the elaboration of sensory qualities. The aim of the present study was to obtain a picture of the dynamics of the microbial ecosystem of RDO Salers cheese by using culture-independent methods. This included DNA extraction, PCR, and single-strand conformation polymorphism (SSCP) analysis. Bacterial and high-GC% gram-positive bacterial primers were used to amplify V2 or V3 regions of the 16S rRNA gene. SSCP patterns revealed changes during the manufacturing of the cheese. Patterns of the ecosystems of cheeses that were provided by three farmers were also quite different. Cloning and sequencing of the 16S rRNA gene revealed sequences related to lactic acid bacteria (Lactococcus lactis, Streptococcus thermophilus, Enterococcus faecium, Leuconostoc mesenteroides, Leuconostoc pseudomesenteroides, Lactobacillus plantarum, and Lactobacillus pentosus), which were predominant during manufacturing and ripening. Bacteria belonging to the high-GC% gram-positive group (essentially corynebacteria) were found by using specific primers. The present molecular approach can effectively describe the ecosystem of artisanal dairy products.
传统奶酪(如法国生产的原产地保护命名的萨勒斯奶酪,一种手工制作的奶酪)在加工和成熟过程中的微生物动态,对其感官品质的形成起着重要作用。本研究的目的是通过使用非培养方法来描绘原产地保护命名的萨勒斯奶酪微生物生态系统的动态。这包括DNA提取、PCR和单链构象多态性(SSCP)分析。使用细菌和高GC%革兰氏阳性细菌引物扩增16S rRNA基因的V2或V3区域。SSCP图谱揭示了奶酪制作过程中的变化。由三位农民提供的奶酪生态系统图谱也有很大差异。16S rRNA基因的克隆和测序揭示了与乳酸菌(乳酸乳球菌、嗜热链球菌、粪肠球菌、肠系膜明串珠菌、类肠系膜明串珠菌、植物乳杆菌和戊糖乳杆菌)相关的序列,这些乳酸菌在制作和成熟过程中占主导地位。使用特异性引物发现了属于高GC%革兰氏阳性菌组(主要是棒状杆菌)的细菌。目前的分子方法可以有效地描述手工乳制品的生态系统。