Bioinformatics Analysis, Prokaryotic Expression and Biological Activity of Lysin from Vibrio parahaemolyticus Phage vB_VpP_1
To study the in vitro cleavage effect of a recombinant lysin from bacteriophage vB_VpP_1 on Vibrio parahaemolyticus, the gp32 gene fragment of vB_VpP_1 was identified as a lysin gene according to the results of whole-genome sequencing (WGS) and functional analysis. The amino acid sequence and struct...
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China Food Publishing Company
2025-02-01
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| Series: | Shipin Kexue |
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| Online Access: | https://www.spkx.net.cn/fileup/1002-6630/PDF/2025-46-3-010.pdf |
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| author | ZHANG Defu, YANG Wenjing, LIU Ke, LIU Qingqing, BAI Wutong, LI Fan, LÜ Xinran, BAI Xue, TAN Xiqian, LI Xuepeng, LI Jianrong |
| author_facet | ZHANG Defu, YANG Wenjing, LIU Ke, LIU Qingqing, BAI Wutong, LI Fan, LÜ Xinran, BAI Xue, TAN Xiqian, LI Xuepeng, LI Jianrong |
| author_sort | ZHANG Defu, YANG Wenjing, LIU Ke, LIU Qingqing, BAI Wutong, LI Fan, LÜ Xinran, BAI Xue, TAN Xiqian, LI Xuepeng, LI Jianrong |
| collection | DOAJ |
| description | To study the in vitro cleavage effect of a recombinant lysin from bacteriophage vB_VpP_1 on Vibrio parahaemolyticus, the gp32 gene fragment of vB_VpP_1 was identified as a lysin gene according to the results of whole-genome sequencing (WGS) and functional analysis. The amino acid sequence and structure of gp32 were analyzed by various tools such as Expasy. Primers were designed using Primer 5.0 software and cloned into the pET-28a(+) vector for prokaryotic expression. The lytic activity of the purified lysin against the host bacterium before and after being treated with ethylene diamine tetraacetic acid (EDTA) was measured. Tertiary structure analysis showed that the lysin was a spherical hydrophilic protein, which was predicted to contain two catalytic domains. This was consistent with the basic characteristics of Gram-negative phage lysins. The lysin had no transmembrane region or signal peptide. The purified bacteriophage vB_VpP_1 had a lysin activity of approximately (1 487 ± 182) U/mg, which showed a strong capacity to lyse V. parahaemolyticus with damaged cell walls but not V. parahaemolyticus with intact cell walls. The prokaryotic expression vector for bacteriophage vB_VpP_1 lysin was successfully constructed, and the expressed and purified lysin could lyse V. parahaemolyticus with damaged cell walls. |
| format | Article |
| id | doaj-art-fb0770b914ad4701a10c7fa6d5b7a515 |
| institution | Kabale University |
| issn | 1002-6630 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | China Food Publishing Company |
| record_format | Article |
| series | Shipin Kexue |
| spelling | doaj-art-fb0770b914ad4701a10c7fa6d5b7a5152025-02-05T09:08:33ZengChina Food Publishing CompanyShipin Kexue1002-66302025-02-01463838910.7506/spkx1002-6630-20240513-101Bioinformatics Analysis, Prokaryotic Expression and Biological Activity of Lysin from Vibrio parahaemolyticus Phage vB_VpP_1ZHANG Defu, YANG Wenjing, LIU Ke, LIU Qingqing, BAI Wutong, LI Fan, LÜ Xinran, BAI Xue, TAN Xiqian, LI Xuepeng, LI Jianrong0(Institute of Ocean Research, National and Local Joint Engineering Research Center of Storage, Processing and Safety Control Technology for Fresh Agricultural and Aquatic Products, Food Safety Key Lab of Liaoning Province, College of Food Science and Engineering, Bohai University, Jinzhou 121013, China)To study the in vitro cleavage effect of a recombinant lysin from bacteriophage vB_VpP_1 on Vibrio parahaemolyticus, the gp32 gene fragment of vB_VpP_1 was identified as a lysin gene according to the results of whole-genome sequencing (WGS) and functional analysis. The amino acid sequence and structure of gp32 were analyzed by various tools such as Expasy. Primers were designed using Primer 5.0 software and cloned into the pET-28a(+) vector for prokaryotic expression. The lytic activity of the purified lysin against the host bacterium before and after being treated with ethylene diamine tetraacetic acid (EDTA) was measured. Tertiary structure analysis showed that the lysin was a spherical hydrophilic protein, which was predicted to contain two catalytic domains. This was consistent with the basic characteristics of Gram-negative phage lysins. The lysin had no transmembrane region or signal peptide. The purified bacteriophage vB_VpP_1 had a lysin activity of approximately (1 487 ± 182) U/mg, which showed a strong capacity to lyse V. parahaemolyticus with damaged cell walls but not V. parahaemolyticus with intact cell walls. The prokaryotic expression vector for bacteriophage vB_VpP_1 lysin was successfully constructed, and the expressed and purified lysin could lyse V. parahaemolyticus with damaged cell walls.https://www.spkx.net.cn/fileup/1002-6630/PDF/2025-46-3-010.pdfvibrio parahaemolyticus; bacteriophage; lysin; biocontrol agents; food safety |
| spellingShingle | ZHANG Defu, YANG Wenjing, LIU Ke, LIU Qingqing, BAI Wutong, LI Fan, LÜ Xinran, BAI Xue, TAN Xiqian, LI Xuepeng, LI Jianrong Bioinformatics Analysis, Prokaryotic Expression and Biological Activity of Lysin from Vibrio parahaemolyticus Phage vB_VpP_1 Shipin Kexue vibrio parahaemolyticus; bacteriophage; lysin; biocontrol agents; food safety |
| title | Bioinformatics Analysis, Prokaryotic Expression and Biological Activity of Lysin from Vibrio parahaemolyticus Phage vB_VpP_1 |
| title_full | Bioinformatics Analysis, Prokaryotic Expression and Biological Activity of Lysin from Vibrio parahaemolyticus Phage vB_VpP_1 |
| title_fullStr | Bioinformatics Analysis, Prokaryotic Expression and Biological Activity of Lysin from Vibrio parahaemolyticus Phage vB_VpP_1 |
| title_full_unstemmed | Bioinformatics Analysis, Prokaryotic Expression and Biological Activity of Lysin from Vibrio parahaemolyticus Phage vB_VpP_1 |
| title_short | Bioinformatics Analysis, Prokaryotic Expression and Biological Activity of Lysin from Vibrio parahaemolyticus Phage vB_VpP_1 |
| title_sort | bioinformatics analysis prokaryotic expression and biological activity of lysin from vibrio parahaemolyticus phage vb vpp 1 |
| topic | vibrio parahaemolyticus; bacteriophage; lysin; biocontrol agents; food safety |
| url | https://www.spkx.net.cn/fileup/1002-6630/PDF/2025-46-3-010.pdf |
| work_keys_str_mv | AT zhangdefuyangwenjingliukeliuqingqingbaiwutonglifanluxinranbaixuetanxiqianlixuepenglijianrong bioinformaticsanalysisprokaryoticexpressionandbiologicalactivityoflysinfromvibrioparahaemolyticusphagevbvpp1 |