de la Plaza M, Rodríguez A, Fernández de Palencia P, Martínez-Cuesta M C, Peláez C, Requena T
Department of Dairy Science and Technology, Instituto del Frío (CSIC), Madrid, Spain.
Lett Appl Microbiol. 2006 Dec;43(6):637-44. doi: 10.1111/j.1472-765X.2006.02012.x.
The use of randomly amplified polymorphic DNA (RAPD)-PCR fingerprinting and plasmid profiles to determine at the strain level, the similarity of Lactococcus lactis isolates obtained during sampling of traditional cheeses and to verify its correspondence to the selected phenotypic characteristics.
A total of 45 L. lactis isolates were genotypically analysed by RAPD-PCR fingerprinting and plasmid patterns. Phenotypic traits used to compare strains were proteolytic, acidifying, aminotransferase (aromatic and branched chain aminotransferase) and alpha-ketoisovalerate decarboxylase (Kivd) activities. The results show that 23 isolates could be grouped in clusters that exhibited 100% identity in both their RAPD and plasmid patterns, indicating the probable isolation of dominant strains during the cheese sampling process. However, there were phenotypic differences between isolates within the same cluster that included the loss of relevant technological properties such as proteinase activity and acidifying capacity or high variation in their amino acid converting enzyme activities. Likewise, the analysis of a specific attribute, Kivd activity, indicated that 7 of 15 isolates showed no detectable activity despite the presence of the encoding (kivd) gene.
Phenotypic differences found between genotypically similar strains of L. lactis strains could be linked to differences in enzymatic expression.
Phenotypic analysis of L. lactis isolates should be considered when selecting strains with new cheese flavour forming capabilities.
利用随机扩增多态性DNA(RAPD)-PCR指纹图谱和质粒图谱,在菌株水平上确定传统奶酪采样过程中获得的乳酸乳球菌分离株的相似性,并验证其与所选表型特征的对应关系。
通过RAPD-PCR指纹图谱和质粒图谱对总共45株乳酸乳球菌分离株进行了基因分型分析。用于比较菌株的表型特征包括蛋白水解、酸化、转氨酶(芳香族和支链转氨酶)和α-酮异戊酸脱羧酶(Kivd)活性。结果表明,23株分离株可归为簇群,它们的RAPD和质粒图谱均显示100%的同一性,这表明在奶酪采样过程中可能分离出了优势菌株。然而,同一簇群内的分离株之间存在表型差异,包括蛋白酶活性和酸化能力等相关工艺特性的丧失,或者其氨基酸转化酶活性的高度变异。同样,对特定属性Kivd活性的分析表明,15株分离株中有7株尽管存在编码(kivd)基因,但未检测到活性。
乳酸乳球菌基因型相似菌株之间发现的表型差异可能与酶表达差异有关。
在选择具有新奶酪风味形成能力的菌株时,应考虑对乳酸乳球菌分离株进行表型分析。