Nie Ting, Meng Fanqiang, Zhou Libang, Lu Fengxia, Bie Xiaomei, Lu Zhaoxin, Lu Yingjian
College of Food Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu Province 210095, China.
College of Food Science and Engineering, Nanjing University of Finance and Economics, Nanjing, Jiangsu Province 210023, China.
J Agric Food Chem. 2021 Mar 31;69(12):3751-3760. doi: 10.1021/acs.jafc.0c07450. Epub 2021 Feb 10.
Four novel chimeric lysins (P361, P362, P371, and P372), which were the fusion of phage lysins and novel antimicrobial peptide LeuA-P, were obtained using bioinformatics analysis and design. The recombinant chimeric lysins were expressed in BL21(DE3) strain and showed highly specific inhibition against . The minimal inhibitory concentrations (MICs) of P362 and P372 to CMCC 50071 were 8 and 16 μg/mL, respectively. Both 1 × MIC P362 and P372 could increase the outer membrane permeability and cleave the cell wall peptidoglycan, causing the leakage of intracellular nucleic acids and proteins and ultimately killing efficiently without drug resistance. The combination of P362, P372, and potassium sorbate reduced more than 3 log CFU/g counts of microorganisms in contaminated chilled chicken and extended the shelf life by 7 days. The strategy of antimicrobial peptide (AMP)-lysin chimera inspired the inability of phage lysin to specifically inhibit Gram-negative bacteria with dense outer membranes .
通过生物信息学分析和设计，获得了四种新型嵌合溶素（P361、P362、P371和P372），它们是噬菌体溶素与新型抗菌肽LeuA-P的融合体。重组嵌合溶素在BL21(DE3)菌株中表达，并对……表现出高度特异性抑制。P362和P372对CMCC 50071的最低抑菌浓度（MIC）分别为8和16μg/mL。1×MIC的P362和P372均可增加外膜通透性并裂解细胞壁肽聚糖，导致细胞内核酸和蛋白质泄漏，最终有效杀灭细菌且无耐药性。P362、P372和山梨酸钾的组合使受污染冰鲜鸡肉中的微生物数量减少了超过3 log CFU/g，并将保质期延长了7天。抗菌肽（AMP）-溶素嵌合体策略克服了噬菌体溶素无法特异性抑制具有致密外膜的革兰氏阴性菌的问题。
