铜绿假单胞菌噬菌体K4的性质及其在食品防腐方面的应用  

Properties of Pseudomonas aeruginosa Phage K4 and Its Applications in Food Preservation

作　　者：庞文静 韩庆竹 尤甲甲 李东航 李佩泽 李玥莹 杨洪江[1] PANG Wenjing;HAN Qingzhu;YOU Jiajia;LI Donghang;LI Peize;LI Yueying;YANG Hongjiang(Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,College of Biotechnology,Tianjin University of Science and Technology,Tianjin 300457,China)

机构地区：[1]工业发酵微生物教育部重点实验室,天津市工业微生物重点实验室,天津科技大学生物工程学院,天津300457

出　　处：《食品工业科技》2022年第16期130-139,共10页Science and Technology of Food Industry

基　　金：国家自然科学基金(31970150)。

摘　　要：目的:为研究噬菌体在控制细菌污染方面的应用,本文对铜绿假单胞菌噬菌体K4的生物学性质及抑菌活性进行了探究。方法:包括一步生长曲线、稳定性研究、基因组测序、比较基因组学分析以及抑菌曲线等方法。结果:K4潜伏期约为15 min,释放量约为85 PFU/infection center,具有较强的感染性;在不同感染复数(MOI)下,噬菌体K4对宿主菌均有明显的抑制效果;基因组分析显示,K4基因组长度为50358 bp,编码77个蛋白和1个tRNA-Arg;比较基因组学分析发现,噬菌体K4与Paundecimvirus属的Pseudomonas virus PA11基因组序列的一致性达95.08%,证实噬菌体K4是该属的新成员。在应用实验中,噬菌体K4能够显著抑制牛奶和午餐肉等样品中宿主菌的生长,同时噬菌体的数量也有显著增加。本研究结果显示,噬菌体K4生长速度快,具有较强的杀菌活性,基因组不携带整合酶、抗性以及毒力因子等编码基因。结论:噬菌体K4可以用于控制食品中铜绿假单胞菌的污染。Objective: To study phage applications in the control of bacteria contaminations, Pseudomonas aeruginosa phage K4 was characterized for its biological characteristics and inhibition effects of bacterial growth. Methods: One-step growth curve, phage stability, genome sequencing, comparative genomics analysis, and growth inhibition curve were conducted in this work. Results: One-step growth curve showed that phage K4 had a latent phase of 15 min and a burst size of about85 PFU/infection center, displaying a strong infectivity. Under different MOIs(multiplicity of infection), phage K4efficiently inhibited the growth of the host cells. Genomic analysis revealed that phage K4 had a genome of 50358 bp with77 coding sequences and 1 tRNA-Arg gene. Comparative genomics showed that the genome of phage K4 shared an identity of 95.08% with Pseudomonas virus PA11 which belongs to the Paundecimvirus genus of the Zobellviridae family,suggesting that phage K4 might be a new member of the Paundecimvirus genus. In application assays, phage K4 could significantly control the growth of the host cells in sterilized milk and ready-to-eat beef samples along with obvious reproductions of phage K4. In conclusion, the data suggested that phage K4 was a lytic virus that replicated rapidly with relatively high bactericidal activity. Conclusion: The genome of phage K4 did not carry genes for integration, antibiotic resistance, and virulence, appropriately to be selected as a biocontrol agent in the control and prevention of P. aeruginosa contaminations in foods.

关 键 词：铜绿假单胞菌 噬菌体 抑菌活性 基因组注释 应用 

分 类 号：Q939.48[生物学—微生物学]