Deutsch Stéphanie Marie, Neveu Anthony, Guezenec Stéphane, Ritzenthaler Paul, Lortal Sylvie
Laboratoire de Recherches de Technologie Laitière, Institut National de la Recherche Agronomique, 65 rue de Saint-Brieuc, 35042 Rennes Cédex, France.
Int J Food Microbiol. 2003 Mar 15;81(2):147-57. doi: 10.1016/s0168-1605(02)00225-8.
Lactobacillus helveticus is mainly used as starter in Swiss-type cheeses. Often, lysogenic strains are eliminated because of the risk of early lysis and acidification failure due to phage expression. On the other hand, L. helveticus lysis was shown to positively influence cheese proteolysis during ripening. In order to better assess the relationship between lysis and lysogeny, a prophage-cured derivative of L. helveticus CNRZ 303 was isolated (LH 303-G11) and relysogenised (LH 303-G11R), as demonstrated by hybridisation using the whole phage DNA as probe. The growth, lysis in buffered solutions and lytic activities in zymogram using either Micrococcus luteus or L. helveticus as substrate were identical between the mother strain and its cured derivatives. Only morphological differences were observed by scanning electron microscopy: the cells of the cured derivative were shorter in length. The mother strain and its cured and relysogenised derivatives were assayed in triplicate in experimental Swiss cheeses (scale 1:100). No differences were noted during the cheese making: the three strains exhibited identical kinetics of acidification, leading to similar cheeses at day 1 in terms of gross composition and pH. Phages were detected only in the cheeses made with the mother strain and the relysogenised derivative. The lysis of L. helveticus, estimated by viability decrease and release of the intracellular marker D-lactate deshydrogenase, started early before brining and continued during the cold room ripening. No obvious differences of lysis extent were observed. These results demonstrated for the first time that, in the case of LH 303, the extensive lysis observed in cheese is mainly due to autolysin activity and not to prophage induction.
瑞士乳杆菌主要用作瑞士型奶酪的发酵剂。由于噬菌体表达存在早期裂解和酸化失败的风险,溶原性菌株常常被去除。另一方面,研究表明瑞士乳杆菌的裂解对奶酪成熟过程中的蛋白质水解有积极影响。为了更好地评估裂解与溶原性之间的关系,分离出了瑞士乳杆菌CNRZ 303的一个前噬菌体清除衍生物(LH 303-G11)并使其再次溶原化(LH 303-G11R),以全噬菌体DNA为探针进行杂交证明了这一点。母菌株与其清除衍生物在缓冲溶液中的生长、裂解以及以藤黄微球菌或瑞士乳杆菌为底物的酶谱中的裂解活性是相同的。通过扫描电子显微镜仅观察到形态学差异:清除衍生物的细胞长度较短。母菌株及其清除和再次溶原化的衍生物在实验瑞士奶酪(比例1:100)中进行了一式三份的测定。在奶酪制作过程中未发现差异:这三种菌株表现出相同的酸化动力学,在第1天就产生了在总体组成和pH方面相似的奶酪。仅在由母菌株和再次溶原化衍生物制作的奶酪中检测到噬菌体。通过活力降低和细胞内标记物D-乳酸脱氢酶的释放来估计的瑞士乳杆菌的裂解在腌制前就早早开始,并在冷藏室成熟期间持续进行。未观察到裂解程度的明显差异。这些结果首次证明,在LH 303的情况下,在奶酪中观察到的广泛裂解主要是由于自溶素活性而非前噬菌体诱导。