Barbosa M S, Todorov S D, Belguesmia Y, Choiset Y, Rabesona H, Ivanova I V, Chobert J-M, Haertlé T, Franco B D G M
Departamento de Alimentos e Nutrição Experimental, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, São Paulo, Brazil.
J Appl Microbiol. 2014 May;116(5):1195-208. doi: 10.1111/jam.12438. Epub 2014 Feb 8.
The study aimed at determining the biochemical characteristics of the bacteriocin produced by Lactobacillus sakei MBSa1, isolated from salami, correlating the results with the genetic features of the producer strain.
Identification of strain MBSa1 was performed by 16S rDNA sequencing. The bacteriocin was tested for spectrum of activity, heat and pH stability, mechanism of action, molecular mass and amino acid sequence when purified by cation-exchange and reversed-phase HPLC. Genomic DNA was tested for bacteriocin genes commonly present in Lact. sakei. Bacteriocin MBSa1 was heat-stable, unaffected by pH 2·0 to 6·0 and active against all tested Listeria monocytogenes strains. Maximal production of bacteriocin MBSa1 (1600 AU ml(-1)) in MRS broth occurred after 20 h at 25°C. The molecular mass of produced bacteriocin was 4303·3 Da, and the molecule contained the SIIGGMISGWAASGLAG sequence, also present in sakacin A. The strain contained the sakacin A and curvacin A genes but was negative for other tested sakacin genes (sakacins T-α, T-β, X, P, G and Q).
In the studied conditions, Lact. sakei MBSa1 produced sakacin A, a class II bacteriocin, with anti-Listeria activity.
The study covers the purification and characterization of the bacteriocin produced by a lactic acid bacteria isolated from salami (Lact. sakei MBSa1), linking genetic and expression information. Its heat-resistance, pH stability in acid conditions (pH 2·0-6·0) and activity against L. monocytogenes food isolates bring up a potential technological application to improve food safety.
本研究旨在确定从意大利腊肠中分离出的清酒乳杆菌MBSa1所产生细菌素的生化特性,并将结果与产生菌菌株的遗传特征相关联。
通过16S rDNA测序对菌株MBSa1进行鉴定。对细菌素进行了活性谱、热稳定性和pH稳定性、作用机制、分子量以及通过阳离子交换和反相高效液相色谱纯化后的氨基酸序列测试。对基因组DNA进行了清酒乳杆菌中常见细菌素基因的检测。细菌素MBSa1具有热稳定性,在pH 2.0至6.0范围内不受影响,且对所有测试的单核细胞增生李斯特菌菌株均有活性。在MRS肉汤中,25°C培养20小时后,细菌素MBSa1的产量最高(1600 AU ml⁻¹)。所产生细菌素的分子量为4303.3 Da,该分子包含SIIGGMISGWAASGLAG序列,此序列也存在于片球菌素A中。该菌株含有片球菌素A和弯曲菌素A基因,但对其他测试的片球菌素基因(片球菌素T-α、T-β、X、P、G和Q)呈阴性。
在研究条件下,清酒乳杆菌MBSa1产生了具有抗李斯特菌活性的II类细菌素片球菌素A。
本研究涵盖了从意大利腊肠中分离出的乳酸菌(清酒乳杆菌MBSa1)所产生细菌素的纯化和特性分析,将遗传信息与表达信息联系起来。其耐热性、在酸性条件下(pH 2.0 - 6.0)的pH稳定性以及对食品分离的单核细胞增生李斯特菌的活性为改善食品安全带来了潜在的技术应用。