Université de Lyon, Lyon, France.
J Appl Microbiol. 2010 Feb;108(2):647-57. doi: 10.1111/j.1365-2672.2009.04459.x. Epub 2009 Jul 7.
The aim was to isolate and characterize Lactococcus strains with new properties compared to those of usual Lactococcus dairy starters derived from cow's milk.
Algerian dromedary's milk was screened for proteolytic isolates able to grow rapidly on agar milk medium. PCR experiments revealed that 74 proteolytic isolates belonged to the genus Lactococcus and harboured the prtP gene encoding the lactococcal cell-surface proteinase. Among these, 85% were able to ferment citrate (Cit(+) phenotype) and were classified as Lactococcus lactis ssp. lactis biovar. diacetylactis. This classification was confirmed after sequencing of the 16S rDNA gene of five Cit(+) isolates. In contrast to dairy lactococci described in the literature, several Cit(+) isolates exhibited a tolerance to 50 degrees C (Ther(+)) and alkaline pH. Two genetic approaches allowed to show the presence of four independent plasmids (so-called pTher, pPrt, pLac, pCit) associated with the four respective phenotypes: Ther(+), cell-surface proteinase activity PrtP (PrtP(+)), lactose catabolism (Lac(+)) and citrate utilization (Cit(+)). Two types of pCit plasmid were amplified by inverse PCR: class 1 was characterized by a 9-kb plasmid harbouring the expected lactococcal citQRP operon and class 2 by a 23-kb plasmid harbouring the Leuconostoc cit cluster (citI-CitMCDEFGRP).
This work enlarges knowledge of the biovariety diacetylactis by far mainly limited to the citrate-fermenting ability and suggests that the cit plasmid system of some lactococcal strains could have been acquired from another lactic acid bacteria (Leuconostoc spp.).
This study reveals new potential dairy lactococci starters of the biovariety diacetylactis able to grow rapidly in milk at a higher temperature in addition to their casein, lactose and citrate-utilizing abilities.
本研究旨在从牛乳来源的常规乳球菌乳制品发酵剂中分离和鉴定具有新特性的乳球菌菌株。
从阿尔及利亚单峰驼乳中筛选可在琼脂乳培养基上快速生长的蛋白酶产生菌。PCR 实验表明,74 株蛋白酶产生菌属于乳球菌属,携带编码乳球菌细胞表面蛋白酶的 prtP 基因。其中,85%的菌株能够发酵柠檬酸盐(Cit(+)表型),被归类为乳球菌乳亚种双乙酰亚种。这一分类在对 5 株 Cit(+)分离株的 16S rDNA 基因测序后得到了确认。与文献中描述的乳制品乳球菌不同,一些 Cit(+)分离株表现出对 50°C(Ther(+))和碱性 pH 的耐受性。两种遗传方法表明,存在与四种表型(Ther(+)、细胞表面蛋白酶活性 PrtP(PrtP(+))、乳糖代谢(Lac(+))和柠檬酸盐利用(Cit(+))相关的四种独立质粒(所谓的 pTher、pPrt、pLac、pCit)。通过反向 PCR 扩增了两种类型的 pCit 质粒:1 类质粒的特征是含有预期的乳球菌 citQRP 操纵子的 9kb 质粒,2 类质粒的特征是含有 Leuconostoc cit 簇(citI-CitMCDEFGRP)的 23kb 质粒。
本研究扩大了对双乙酰亚种的认识,迄今为止,该亚种主要限于柠檬酸发酵能力,并表明一些乳球菌菌株的 cit 质粒系统可能是从另一种乳酸菌(肠膜明串珠菌属)获得的。
本研究揭示了新的潜在的乳制品发酵剂乳球菌双乙酰亚种,除了能够快速利用酪蛋白、乳糖和柠檬酸盐外,还能在更高的温度下在牛奶中快速生长。