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>...
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2020-12-01
|
Series: | Processes |
Subjects: | |
Online Access: | https://www.mdpi.com/2227-9717/8/12/1587 |
id |
doaj-f779aeb7e79f4cac9042f2da2b56e7c7 |
---|---|
record_format |
Article |
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 |
work_keys_str_mv |
AT fengwang designoverproductionandpurificationofthechimericphagelysinmltphgfightingagainstistaphylococcusaureusi AT xiaohangliu designoverproductionandpurificationofthechimericphagelysinmltphgfightingagainstistaphylococcusaureusi AT zhengyudeng designoverproductionandpurificationofthechimericphagelysinmltphgfightingagainstistaphylococcusaureusi AT yaozhang designoverproductionandpurificationofthechimericphagelysinmltphgfightingagainstistaphylococcusaureusi AT xinyuji designoverproductionandpurificationofthechimericphagelysinmltphgfightingagainstistaphylococcusaureusi AT yanxiong designoverproductionandpurificationofthechimericphagelysinmltphgfightingagainstistaphylococcusaureusi AT lianbinglin designoverproductionandpurificationofthechimericphagelysinmltphgfightingagainstistaphylococcusaureusi |
_version_ |
1724410677562441728 |