Design, Overproduction and Purification of the Chimeric Phage Lysin MLTphg Fighting against <i>Staphylococcus aureus</i>

With the increasing spread of multidrug-resistant bacterial pathogens, it is of great importance to develop alternatives to conventional antibiotics. Here, we report the generation of a chimeric phage lysin, MLTphg, which was assembled by joining the lysins derived from <i>Meiothermus</i>...

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Main Authors: Feng Wang, Xiaohang Liu, Zhengyu Deng, Yao Zhang, Xinyu Ji, Yan Xiong, Lianbing Lin
Format: Article
Language:English
Published: MDPI AG 2020-12-01
Series:Processes
Subjects:
Online Access:https://www.mdpi.com/2227-9717/8/12/1587
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spelling doaj-f779aeb7e79f4cac9042f2da2b56e7c72020-12-02T00:01:30ZengMDPI AGProcesses2227-97172020-12-0181587158710.3390/pr8121587Design, Overproduction and Purification of the Chimeric Phage Lysin MLTphg Fighting against <i>Staphylococcus aureus</i>Feng Wang0Xiaohang Liu1Zhengyu Deng2Yao Zhang3Xinyu Ji4Yan Xiong5Lianbing Lin6Faculty of Life Science and Technology, Kunming University of Science and Technology, 727 South Jingming Road, Kunming 650500, ChinaFaculty of Life Science and Technology, Kunming University of Science and Technology, 727 South Jingming Road, Kunming 650500, ChinaFaculty of Life Science and Technology, Kunming University of Science and Technology, 727 South Jingming Road, Kunming 650500, ChinaFaculty of Life Science and Technology, Kunming University of Science and Technology, 727 South Jingming Road, Kunming 650500, ChinaFaculty of Life Science and Technology, Kunming University of Science and Technology, 727 South Jingming Road, Kunming 650500, ChinaFaculty of Life Science and Technology, Kunming University of Science and Technology, 727 South Jingming Road, Kunming 650500, ChinaFaculty of Life Science and Technology, Kunming University of Science and Technology, 727 South Jingming Road, Kunming 650500, ChinaWith the increasing spread of multidrug-resistant bacterial pathogens, it is of great importance to develop alternatives to conventional antibiotics. Here, we report the generation of a chimeric phage lysin, MLTphg, which was assembled by joining the lysins derived from <i>Meiothermus</i> bacteriophage MMP7 and <i>Thermus</i> bacteriophage TSP4 with a flexible linker via chimeolysin engineering. As a potential antimicrobial agent, MLTphg can be obtained by overproduction in <i>Escherichia coli</i> BL21(DE3) cells and the following Ni-affinity chromatography. Finally, we recovered about 40 ± 1.9 mg of MLTphg from 1 L of the host <i>E. coli</i> BL21(DE3) culture. The purified MLTphg showed peak activity against <i>Staphylococcus aureus</i> ATCC6538 between 35 and 40 °C, and maintained approximately 44.5 ± 2.1% activity at room temperature (25 °C). Moreover, as a produced chimera, it exhibited considerably improved bactericidal activity against <i>Staphylococcus aureus</i> (2.9 ± 0.1 log<sub>10</sub> reduction was observed upon 40 nM MLTphg treatment at 37 °C for 30 min) and also a group of antibiotic-resistant bacteria compared to its parental lysins, TSPphg and MMPphg. In the current age of growing antibiotic resistance, our results provide an engineering basis for developing phage lysins as novel antimicrobial agents and shed light on bacteriophage-based strategies to tackle bacterial infections.https://www.mdpi.com/2227-9717/8/12/1587bacteriophageendolysinbioengineeringpathogen control<i>Staphylococcus aureus</i>antibiotic-resistant bacteria
collection DOAJ
language English
format Article
sources DOAJ
author Feng Wang
Xiaohang Liu
Zhengyu Deng
Yao Zhang
Xinyu Ji
Yan Xiong
Lianbing Lin
spellingShingle Feng Wang
Xiaohang Liu
Zhengyu Deng
Yao Zhang
Xinyu Ji
Yan Xiong
Lianbing Lin
Design, Overproduction and Purification of the Chimeric Phage Lysin MLTphg Fighting against <i>Staphylococcus aureus</i>
Processes
bacteriophage
endolysin
bioengineering
pathogen control
<i>Staphylococcus aureus</i>
antibiotic-resistant bacteria
author_facet Feng Wang
Xiaohang Liu
Zhengyu Deng
Yao Zhang
Xinyu Ji
Yan Xiong
Lianbing Lin
author_sort Feng Wang
title Design, Overproduction and Purification of the Chimeric Phage Lysin MLTphg Fighting against <i>Staphylococcus aureus</i>
title_short Design, Overproduction and Purification of the Chimeric Phage Lysin MLTphg Fighting against <i>Staphylococcus aureus</i>
title_full Design, Overproduction and Purification of the Chimeric Phage Lysin MLTphg Fighting against <i>Staphylococcus aureus</i>
title_fullStr Design, Overproduction and Purification of the Chimeric Phage Lysin MLTphg Fighting against <i>Staphylococcus aureus</i>
title_full_unstemmed Design, Overproduction and Purification of the Chimeric Phage Lysin MLTphg Fighting against <i>Staphylococcus aureus</i>
title_sort design, overproduction and purification of the chimeric phage lysin mltphg fighting against <i>staphylococcus aureus</i>
publisher MDPI AG
series Processes
issn 2227-9717
publishDate 2020-12-01
description With the increasing spread of multidrug-resistant bacterial pathogens, it is of great importance to develop alternatives to conventional antibiotics. Here, we report the generation of a chimeric phage lysin, MLTphg, which was assembled by joining the lysins derived from <i>Meiothermus</i> bacteriophage MMP7 and <i>Thermus</i> bacteriophage TSP4 with a flexible linker via chimeolysin engineering. As a potential antimicrobial agent, MLTphg can be obtained by overproduction in <i>Escherichia coli</i> BL21(DE3) cells and the following Ni-affinity chromatography. Finally, we recovered about 40 ± 1.9 mg of MLTphg from 1 L of the host <i>E. coli</i> BL21(DE3) culture. The purified MLTphg showed peak activity against <i>Staphylococcus aureus</i> ATCC6538 between 35 and 40 °C, and maintained approximately 44.5 ± 2.1% activity at room temperature (25 °C). Moreover, as a produced chimera, it exhibited considerably improved bactericidal activity against <i>Staphylococcus aureus</i> (2.9 ± 0.1 log<sub>10</sub> reduction was observed upon 40 nM MLTphg treatment at 37 °C for 30 min) and also a group of antibiotic-resistant bacteria compared to its parental lysins, TSPphg and MMPphg. In the current age of growing antibiotic resistance, our results provide an engineering basis for developing phage lysins as novel antimicrobial agents and shed light on bacteriophage-based strategies to tackle bacterial infections.
topic bacteriophage
endolysin
bioengineering
pathogen control
<i>Staphylococcus aureus</i>
antibiotic-resistant bacteria
url https://www.mdpi.com/2227-9717/8/12/1587
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