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Efficient expression of sortase A from Staphylococcus aureus in Escherichia coli and its enzymatic characterizations
BACKGROUND: Sortase A (SrtA) is a transpeptidase found in Staphylococcus aureus, which is widely used in site-specific protein modification. However, SrtA was expressed in Escherichia coli (E. coli) in rather low level (ranging from several milligrams to 76.9 mg/L at most). The present study aims to...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5316389/ https://www.ncbi.nlm.nih.gov/pubmed/28261538 http://dx.doi.org/10.1186/s40643-017-0143-y |
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author | Wu, Zhimeng Hong, Haofei Zhao, Xinrui Wang, Xun |
author_facet | Wu, Zhimeng Hong, Haofei Zhao, Xinrui Wang, Xun |
author_sort | Wu, Zhimeng |
collection | PubMed |
description | BACKGROUND: Sortase A (SrtA) is a transpeptidase found in Staphylococcus aureus, which is widely used in site-specific protein modification. However, SrtA was expressed in Escherichia coli (E. coli) in rather low level (ranging from several milligrams to 76.9 mg/L at most). The present study aims to optimize fermentation conditions for improving SrtA expression in E. coli. RESULTS: Under the optimized media (0.48 g/L glycerol, 1.37 g/L tryptone, 0.51 g/L yeast extract, MOPS 0.5 g/L, PBS buffer 180 mL/L) and condition (30 °C for 8 h) in a 7-L fermentor, the enzyme activity and the yield of SrtA reached 2458.4 ± 115.9 U/mg DCW and 232.4 ± 21.1 mg/L, respectively, which were higher by 5.8- and 4.5-folds compared with initial conditions, respectively. The yield of SrtA also represented threefold increase than the previously reported maximal level. In addition, the enzymatic characterizations of SrtA (optimal temperature, optimal pH, the influence of metal irons, and tolerance to water-soluble organic solvents) were determined. CONCLUSIONS: Enhanced expression of SrtA was achieved by optimization of medium and condition. This result will have potential application for production levels of SrtA on an industry scale. Moreover, the detailed enzymatic characterizations of SrtA were examined, which will provide a useful guide for its future application. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40643-017-0143-y) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5316389 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-53163892017-03-03 Efficient expression of sortase A from Staphylococcus aureus in Escherichia coli and its enzymatic characterizations Wu, Zhimeng Hong, Haofei Zhao, Xinrui Wang, Xun Bioresour Bioprocess Research BACKGROUND: Sortase A (SrtA) is a transpeptidase found in Staphylococcus aureus, which is widely used in site-specific protein modification. However, SrtA was expressed in Escherichia coli (E. coli) in rather low level (ranging from several milligrams to 76.9 mg/L at most). The present study aims to optimize fermentation conditions for improving SrtA expression in E. coli. RESULTS: Under the optimized media (0.48 g/L glycerol, 1.37 g/L tryptone, 0.51 g/L yeast extract, MOPS 0.5 g/L, PBS buffer 180 mL/L) and condition (30 °C for 8 h) in a 7-L fermentor, the enzyme activity and the yield of SrtA reached 2458.4 ± 115.9 U/mg DCW and 232.4 ± 21.1 mg/L, respectively, which were higher by 5.8- and 4.5-folds compared with initial conditions, respectively. The yield of SrtA also represented threefold increase than the previously reported maximal level. In addition, the enzymatic characterizations of SrtA (optimal temperature, optimal pH, the influence of metal irons, and tolerance to water-soluble organic solvents) were determined. CONCLUSIONS: Enhanced expression of SrtA was achieved by optimization of medium and condition. This result will have potential application for production levels of SrtA on an industry scale. Moreover, the detailed enzymatic characterizations of SrtA were examined, which will provide a useful guide for its future application. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40643-017-0143-y) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2017-02-18 2017 /pmc/articles/PMC5316389/ /pubmed/28261538 http://dx.doi.org/10.1186/s40643-017-0143-y Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Research Wu, Zhimeng Hong, Haofei Zhao, Xinrui Wang, Xun Efficient expression of sortase A from Staphylococcus aureus in Escherichia coli and its enzymatic characterizations |
title | Efficient expression of sortase A from Staphylococcus aureus in Escherichia coli and its enzymatic characterizations |
title_full | Efficient expression of sortase A from Staphylococcus aureus in Escherichia coli and its enzymatic characterizations |
title_fullStr | Efficient expression of sortase A from Staphylococcus aureus in Escherichia coli and its enzymatic characterizations |
title_full_unstemmed | Efficient expression of sortase A from Staphylococcus aureus in Escherichia coli and its enzymatic characterizations |
title_short | Efficient expression of sortase A from Staphylococcus aureus in Escherichia coli and its enzymatic characterizations |
title_sort | efficient expression of sortase a from staphylococcus aureus in escherichia coli and its enzymatic characterizations |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5316389/ https://www.ncbi.nlm.nih.gov/pubmed/28261538 http://dx.doi.org/10.1186/s40643-017-0143-y |
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