Bermúdez-Humarán Luis G, Rihakova Jitka, Langella Philippe, Demnerova Katerina, Nazef Lila, Prévost Hervé, Drider Djamel
Unité d'Ecologie et de Physiologie du Système Digestif, INRA, Domaine de Vilvert, Jouy-en-Josas, France.
J Mol Microbiol Biotechnol. 2007;13(4):259-63. doi: 10.1159/000104756.
Divercin V41 is a class IIa bacteriocin produced by Carnobacterium divergens V41 with a strong anti-Listeria activity. We have previously produced a recombinant form of divercin V41 (DvnRV41) in Escherichia coli strain Origami, by cloning a synthetic gene that codes for a mature divercin RV41 peptide. In this work we describe the inducible expression and secretion of DvnRV41 in the food-grade lactic acid bacterium, Lactococcus lactis. The production of DvnRV41 by recombinant L. lactis was confirmed and quantified by Western blot and ELISA assays. In addition, anti-Listeria activity of DvnRV41 was determined using an agar diffusion test. Although the levels of DvnRV41 produced by recombinant L. lactis were similar to those produced by the natural host, C. divergens V41, the specific activities were lower. In conclusion, our data show that the bacteriocin DvnRV41 is produced and secreted in an active form by L. lactis and that this approach may have important applications in the preservation of foods.
双歧菌素V41是由分歧肉杆菌V41产生的一种IIa类细菌素,具有很强的抗李斯特菌活性。我们之前通过克隆编码成熟双歧菌素RV41肽的合成基因,在大肠杆菌Origami菌株中生产出了重组双歧菌素V41(DvnRV41)。在这项研究中,我们描述了DvnRV41在食品级乳酸菌乳酸乳球菌中的诱导表达和分泌情况。通过蛋白质免疫印迹法和酶联免疫吸附测定法对重组乳酸乳球菌产生的DvnRV41进行了确认和定量。此外,使用琼脂扩散试验测定了DvnRV41的抗李斯特菌活性。尽管重组乳酸乳球菌产生的DvnRV41水平与天然宿主分歧肉杆菌V41产生的水平相似,但其比活性较低。总之,我们的数据表明,细菌素DvnRV41由乳酸乳球菌以活性形式产生并分泌,并且这种方法在食品保鲜中可能具有重要应用。