Borrero Juan, Chen Yuqing, Dunny Gary M, Kaznessis Yiannis N
†Department of Chemical Engineering and Materials Science, ‡Department of Microbiology, University of Minnesota, Minneapolis, Minnesota 55455, United States.
ACS Synth Biol. 2015 Mar 20;4(3):299-306. doi: 10.1021/sb500090b. Epub 2014 Jun 16.
We designed Lactococcus lactis to detect Enterococcus faecalis. Upon detection, L. lactis produce and secrete antienterococcal peptides. The peptides inhibit enterococcal growth and reduce viability of enterococci in the vicinity of L. lactis. The enterococcal sex pheromone cCF10 serves as the signal for detection. Expression vectors derived from pCF10, a cCF10-responsive E. faecalis sex-pheromone conjugative plasmid, were engineered in L. lactis for the detection system. Recombinant host strains were engineered to express genes for three bacteriocins, enterocin A, hiracin JM79 and enterocin P, each with potent antimicrobial activity against E. faecalis. Sensitive detection and specific inhibition occur both in agar and liquid media. The engineered L. lactis also inhibited growth of multidrug-resistant E. faecium strains, when induced by cCF10. The presented vectors and strains can be components of a toolbox for the development of alternative antibiotic technologies targeting enterococci at the site of infection.
我们设计了乳酸乳球菌来检测粪肠球菌。一旦检测到，乳酸乳球菌就会产生并分泌抗肠球菌肽。这些肽会抑制肠球菌的生长，并降低乳酸乳球菌附近肠球菌的活力。肠球菌性信息素cCF10作为检测信号。来自pCF10（一种对cCF10有反应的粪肠球菌性信息素接合质粒）的表达载体在乳酸乳球菌中构建用于检测系统。重组宿主菌株被构建以表达三种细菌素的基因，即肠球菌素A、海拉菌素JM79和肠球菌素P，每种对粪肠球菌都有强大的抗菌活性。在琼脂和液体培养基中都能进行灵敏检测和特异性抑制。当由cCF10诱导时，工程化的乳酸乳球菌也能抑制多重耐药性屎肠球菌菌株的生长。所展示的载体和菌株可以成为在感染部位针对肠球菌开发替代抗生素技术的工具箱的组成部分。
