食品科学 ›› 2026, Vol. 47 ›› Issue (9): 128-137.doi: 10.7506/spkx1002-6630-20250923-183

• 生物工程 • 上一篇    下一篇

4 个噬菌体裂解酶的克隆表达及活性分析

叶文鑫,何嘉欣,Vivian Montero ALEJO,杨美艳   

  1. (1.华南农业大学食品学院,广东 广州 510642;2.古巴药物研究开发中心,古巴 哈瓦那 10100)
  • 出版日期:2026-05-15 发布日期:2026-06-03
  • 基金资助:
    国家自然科学基金青年科学基金项目(32202194);广州市重点研发计划项目(SL2022B03J01243)

Cloning, Expression and Activity Analysis of Four Bacteriophage Lysins

YE Wenxin, HE Jiaxin, Vivian Montero ALEJO, YANG Meiyan   

  1. (1. College of Food Science, South China Agricultural University, Guangzhou 510642, China;2. Cuban Center for Pharmaceutical Research and Development, Havana 10100, Cuba)
  • Online:2026-05-15 Published:2026-06-03

摘要: 基于14 株新型烈性噬菌体(10 株革兰氏阴性菌噬菌体,4 株革兰氏阳性菌噬菌体)挖掘15 个裂解酶基因并对其进行序列分析,通过大肠杆菌BL21(DE3)异源表达系统,最终获得副溶血性弧菌噬菌体裂解酶LysCA8和LysBA3的纯化蛋白以及蜡样芽孢杆菌噬菌体裂解酶LysLY1和LysLY3的粗酶蛋白。其中,裂解酶LysLY1和LysLY3可高效广谱裂解供试的全部89 株蜡样芽孢杆菌,其宿主范围远大于其母体噬菌体(vB_BceS_LY1和vB_BceP_LY3分别仅能裂解其中的3 株和2 株)。裂解酶LysCA8和LysBA3在体外对供试的副溶血性弧菌无裂解活性,推测由环境适应性障碍及单域结构所致。针对革兰氏阴性菌裂解酶无活性的问题,未来将通过结构优化、分子伴侣共表达及嵌合酶设计,揭示失活机制并拓展裂解酶抗菌谱。

关键词: 噬菌体;裂解酶;基因表达;副溶血性弧菌;蜡样芽孢杆菌

Abstract: This study identified and sequenced 15 lysin genes from 14 novel virulent phages (10 Gram-negative and 4 Gram-positive bacterial phages). Through heterologous expression in Escherichia coli BL21(DE3), two phage lysins from Vibrio parahaemolyticus, LysCA8 and LysBA3, were obtained and purified, and two phage lysins from Bacillus cereus, LysLY1 and LysLY3, were also obtained without further purification. LysLY1 and LysLY3 had strong lytic activity against all 89 B. cereus strains tested, demonstrating a host range significantly larger than that of their parent phages (vB_BceS_LY1 and vB_BceP_LY3 were only able to lyse 3 and 2 of these B. cereus strains, respectively). LysCA8 and LysBA3 showed no lytic activity against V. parahaemolyticus in vitro, presumably due to environmental acclimation and their single-domain structure. To address this, future work should focus on structural optimization, co-expression of molecular chaperones, and chimeric enzyme design to uncover the inactivation mechanism and expand the antimicrobial spectrum of these lysins.

Key words: bacteriophage; lysin; gene expression; Vibrio parahaemolyticus; Bacillus cereus

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