[1]沈元朝,李悦,孙利厂,等.噬菌体裂解酶的钝顶螺旋藻表达方法建立及功能评价[J].江苏农业学报,2025,(08):1638-1646.[doi:doi:10.3969/j.issn.1000-4440.2025.08.020]
　SHEN Yuanzhao,LI Yue,SUN Lichang,et al.Establishment of the Spirulina platensis expression method for bacteriophage lysins and functional evaluation[J].,2025,(08):1638-1646.[doi:doi:10.3969/j.issn.1000-4440.2025.08.020]
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噬菌体裂解酶的钝顶螺旋藻表达方法建立及功能评价()

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2025年08期
页码:
1638-1646
栏目:
加工贮藏·质量安全
出版日期:
2025-08-31

文章信息/Info

Title:
Establishment of the Spirulina platensis expression method for bacteriophage lysins and functional evaluation
作者:
沈元朝; 李悦; 孙利厂; 王冉
(江苏省农业科学院农产品质量安全与营养研究所,江苏南京210014)
Author(s):
SHEN Yuanzhao; LI Yue; SUN Lichang; WANG Ran
(Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China)
关键词:
噬菌体裂解酶; 钝顶螺旋藻; 表达方法; 功能评价
Keywords:
bacteriophage lysin; Spirulina platensis; expression method; functional evaluation
分类号:
S188+.2
DOI:
doi:10.3969/j.issn.1000-4440.2025.08.020
文献标志码:
A
摘要:
噬菌体裂解酶因具有高效、安全的杀菌特性成为潜在的抗菌药物,受到研究者的广泛关注。基于本研究团队前期分离得到的1株具有较宽宿主谱且呈烈性的金黄色葡萄球菌噬菌体vB_SauH_JAAS17,本研究首先通过大肠杆菌原核表达系统成功对噬菌体vB_SauH_JAAS17的2种裂解酶——裂解酶A(LysinA)和裂解酶B(LysinB)进行表达分析,证明裂解酶编码基因能在原核表达系统中成功表达和制备,并且将所表达的裂解酶合用时具有广谱杀菌活性。在此基础上,本研究开发了一种基于钝顶螺旋藻表达系统的噬菌体裂解酶的新表达方法。结果显示,在500 ng/μL质粒、1%聚乙二醇8000(PEG 8000)冰浴诱导30 min的转化条件下,LysinA和LysinB的表达水平最高,分别为10.44 mg/mL、10.04 mg/mL。当联合使用的2种噬菌体裂解酶的质量浓度为2.0 mg/mL时,其裂解水平与噬菌体vB_SauH_JAAS17所产裂解酶不存在显著差异,且无明显细胞毒性。
Abstract:
Bacteriophage lysins have emerged as promising antibacterial agents due to their efficient and safe bactericidal properties, attracting widespread attention. In the early stage of this study, a virulent Staphylococcus aureus phage, vB_SauH_JAAS17, with a broad host range was isolated. Based on this, the lysins named LysinA and LysinB encoded by vB_SauH_JAAS17 were successfully expressed using an Escherichia coli prokaryotic expression system, demonstrating both the feasibility of heterologous expression and the broad-spectrum antibacterial activity of the lysins. Furthermore, a novel expression strategy was developed using Spirulina platensis as the host. The results showed that the expression levels of LysinA and LysinB reached their highest values at 10.44 mg/mL and 10.04 mg/mL under transformation conditions of 500 ng/μL plasmid and 1% polyethylene glycol 8000 (PEG 8000) ice bath induction for 30 min. When the mass concentration of the two combined phage lysins was 2.0 mg/mL, their lytic activity showed no significant difference compared to the lysin produced by phage vB_SauH_JAAS17, and no obvious cytotoxicity was observed.

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备注/Memo

备注/Memo:
收稿日期:2025-02-26基金项目:江苏省卓越博士后计划项目(2023ZB375)；江苏省农业科技自主创新基金项目[CX(22)2017、CX(2024)1015]作者简介:沈元朝(1994-),男,江苏南京人,博士,助理研究员,主要从事噬菌体相关生物学技术开发工作。(E-mail)silenceyz525@163.com通讯作者:孙利厂,(E-mail)sunlc@jaas.ac.cn；王冉,(E-mail)ranwang@jaas.ac.cn
更新日期/Last Update: 2025-09-23