Technical data of heterologous expression and purification of SARS-CoV-2 proteases using escherichia coli system
The SARS-CoV-2 coronavirus expresses two essential proteases: firstly, the 3Chymotrypsin-like protease (3CLpro) or main protease (Mpro), and secondly, the papain-like protease (PLpro), both of which are considered as viable drug targets for the inhibition of viral replication. In order to perform dr...
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| Format: | Article |
| Language: | English English |
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MDPI AG
2021
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| Online Access: | https://eprints.ums.edu.my/id/eprint/31419/1/Technical%20data%20of%20heterologous%20expression%20and%20purification%20of%20SARS-CoV-2%20proteases%20using%20escherichia%20coli%20system.pdf https://eprints.ums.edu.my/id/eprint/31419/2/Technical%20data%20of%20heterologous%20expression%20and%20purification%20of%20SARS-CoV-2%20proteases%20using%20escherichia%20coli%20system_ABSTRACT.pdf |
| _version_ | 1825714513431756800 |
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| author | Rafida Razali Vijay Kumar Subbiah Cahyo Budiman |
| author_facet | Rafida Razali Vijay Kumar Subbiah Cahyo Budiman |
| author_sort | Rafida Razali |
| collection | UMS |
| description | The SARS-CoV-2 coronavirus expresses two essential proteases: firstly, the 3Chymotrypsin-like protease (3CLpro) or main protease (Mpro), and secondly, the papain-like protease (PLpro), both of which are considered as viable drug targets for the inhibition of viral replication. In order to perform drug discovery assays for SARS-CoV-2, it is imperative that efficient methods are established for the production and purification of 3CLpro and PLpro of SARS-CoV-2, designated as 3CLpro-CoV2 and PLpro-CoV2, respectively. This article expands the data collected in the attempts to express SARS-CoV-2 proteases under different conditions and purify them under single-step chromatography. Data showed that the use of E. coli BL21(DE3) strain was sufficient to express 3CLpro-CoV2 in a fully soluble form. Nevertheless, the single affinity chromatography step was only applicable for 3CLpro-CoV2 expressed at 18 °C, with a yield and purification fold of 92% and 49, respectively. Meanwhile, PLpro-CoV2 was successfully expressed in a fully soluble form in either BL21(DE3) or BL21-CodonPlus(DE3) strains. In contrast, the single affinity chromatography step was only applicable for PLpro-CoV2 expressed using E. coli BL21-CodonPlus(DE3) at 18 or 37 °C, with a yield and purification fold of 86% (18 °C) or 83.36% (37 °C) and 112 (18 °C) or 71 (37 °C), respectively. The findings provide a guide for optimizing the production of SARS-CoV-2 proteases of E. coli host cells. |
| first_indexed | 2024-03-06T03:12:55Z |
| format | Article |
| id | ums.eprints-31419 |
| institution | Universiti Malaysia Sabah |
| language | English English |
| last_indexed | 2024-03-06T03:12:55Z |
| publishDate | 2021 |
| publisher | MDPI AG |
| record_format | dspace |
| spelling | ums.eprints-314192021-12-13T07:32:57Z https://eprints.ums.edu.my/id/eprint/31419/ Technical data of heterologous expression and purification of SARS-CoV-2 proteases using escherichia coli system Rafida Razali Vijay Kumar Subbiah Cahyo Budiman QR355-502 Virology The SARS-CoV-2 coronavirus expresses two essential proteases: firstly, the 3Chymotrypsin-like protease (3CLpro) or main protease (Mpro), and secondly, the papain-like protease (PLpro), both of which are considered as viable drug targets for the inhibition of viral replication. In order to perform drug discovery assays for SARS-CoV-2, it is imperative that efficient methods are established for the production and purification of 3CLpro and PLpro of SARS-CoV-2, designated as 3CLpro-CoV2 and PLpro-CoV2, respectively. This article expands the data collected in the attempts to express SARS-CoV-2 proteases under different conditions and purify them under single-step chromatography. Data showed that the use of E. coli BL21(DE3) strain was sufficient to express 3CLpro-CoV2 in a fully soluble form. Nevertheless, the single affinity chromatography step was only applicable for 3CLpro-CoV2 expressed at 18 °C, with a yield and purification fold of 92% and 49, respectively. Meanwhile, PLpro-CoV2 was successfully expressed in a fully soluble form in either BL21(DE3) or BL21-CodonPlus(DE3) strains. In contrast, the single affinity chromatography step was only applicable for PLpro-CoV2 expressed using E. coli BL21-CodonPlus(DE3) at 18 or 37 °C, with a yield and purification fold of 86% (18 °C) or 83.36% (37 °C) and 112 (18 °C) or 71 (37 °C), respectively. The findings provide a guide for optimizing the production of SARS-CoV-2 proteases of E. coli host cells. MDPI AG 2021 Article PeerReviewed text en https://eprints.ums.edu.my/id/eprint/31419/1/Technical%20data%20of%20heterologous%20expression%20and%20purification%20of%20SARS-CoV-2%20proteases%20using%20escherichia%20coli%20system.pdf text en https://eprints.ums.edu.my/id/eprint/31419/2/Technical%20data%20of%20heterologous%20expression%20and%20purification%20of%20SARS-CoV-2%20proteases%20using%20escherichia%20coli%20system_ABSTRACT.pdf Rafida Razali and Vijay Kumar Subbiah and Cahyo Budiman (2021) Technical data of heterologous expression and purification of SARS-CoV-2 proteases using escherichia coli system. Data, 6. pp. 1-12. ISSN 2306-5729 https://www.mdpi.com/2306-5729/6/9/99/htm https://doi.org/10.3390/data6090099 https://doi.org/10.3390/data6090099 |
| spellingShingle | QR355-502 Virology Rafida Razali Vijay Kumar Subbiah Cahyo Budiman Technical data of heterologous expression and purification of SARS-CoV-2 proteases using escherichia coli system |
| title | Technical data of heterologous expression and purification of SARS-CoV-2 proteases using escherichia coli system |
| title_full | Technical data of heterologous expression and purification of SARS-CoV-2 proteases using escherichia coli system |
| title_fullStr | Technical data of heterologous expression and purification of SARS-CoV-2 proteases using escherichia coli system |
| title_full_unstemmed | Technical data of heterologous expression and purification of SARS-CoV-2 proteases using escherichia coli system |
| title_short | Technical data of heterologous expression and purification of SARS-CoV-2 proteases using escherichia coli system |
| title_sort | technical data of heterologous expression and purification of sars cov 2 proteases using escherichia coli system |
| topic | QR355-502 Virology |
| url | https://eprints.ums.edu.my/id/eprint/31419/1/Technical%20data%20of%20heterologous%20expression%20and%20purification%20of%20SARS-CoV-2%20proteases%20using%20escherichia%20coli%20system.pdf https://eprints.ums.edu.my/id/eprint/31419/2/Technical%20data%20of%20heterologous%20expression%20and%20purification%20of%20SARS-CoV-2%20proteases%20using%20escherichia%20coli%20system_ABSTRACT.pdf |
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