Popović Nikola, Veljović Katarina, Radojević Dušan, Brdarić Emilija, Stevanović Dušan, Živković Milica, Kojić Milan
Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, 11042 Belgrade, Serbia.
Department of Research and Development, Institute of Virology, Vaccines, and Sera "Torlak", Vojvode Stepe 458, 11152 Belgrade, Serbia.
Foods. 2024 Aug 22;13(16):2637. doi: 10.3390/foods13162637.
This study aimed to investigate the probiogenomic features of artisanal bacteriocin-producing BGPAS1-3 and the use of the improved pMALc5HisEk expression vector for overexpressing class II bacteriocins and the application of purified bacteriocin 31 in a milk model as a preservative against . The BGPAS1-3 strain was isolated from traditional fresh soft cheese manufactured in households on a small scale in rural locations surrounding Pale Mountain City in Bosnia and Herzegovina. The whole-genome sequencing approach and bioinformatics analyses revealed that the strain BGPAS1-3 was non-pathogenic to humans. The presence of bacteriocin operons suggested the ability of the isolate to suppress the growth of pathogens. Coding regions for three maturated bacteriocins (bacteriocin 31, bacteriocin 32, and enterocin P) produced by BGPAS1-3 were amplified and expressed in ER2523 using the pMALc5HisEk system. All three bacteriocins were successfully overexpressed and purified after enterokinase cleavage but showed different antimicrobial activity. Bacteriocin 31 showed significantly stronger antimicrobial activity compared with bacteriocin 32. It was the only one that proved to be suitable for use as a food preservative against in a milk model.
本研究旨在探究手工制作的产细菌素的BGPAS1-3的益生菌基因组特征,以及使用改良的pMALc5HisEk表达载体过表达II类细菌素,并将纯化的细菌素31作为防腐剂应用于牛奶模型中以抵抗……。BGPAS1-3菌株是从波斯尼亚和黑塞哥维那帕莱山城周边农村地区家庭小规模制作的传统新鲜软奶酪中分离出来的。全基因组测序方法和生物信息学分析表明,BGPAS1-3菌株对人类无致病性。细菌素操纵子的存在表明该分离株具有抑制病原体生长的能力。利用pMALc5HisEk系统,扩增了BGPAS1-3产生的三种成熟细菌素(细菌素31、细菌素32和肠球菌素P)的编码区,并在ER2523中进行表达。经肠激酶切割后,所有三种细菌素均成功过表达并纯化,但表现出不同的抗菌活性。与细菌素32相比,细菌素31表现出显著更强的抗菌活性。它是唯一被证明适用于牛奶模型中作为抵抗……的食品防腐剂的细菌素。