Impact of the Expression System on Recombinant Protein Production in Escherichia coli BL21
Recombinant protein production for medical, academic, or industrial applications is essential for our current life. Recombinant proteins are obtained mainly through microbial fermentation, with Escherichia coli being the host most used. In spite of that, some problems are associated with the product...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2021-06-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2021.682001/full |
| _version_ | 1819043969589313536 |
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| author | Gema Lozano Terol Julia Gallego-Jara Rosa Alba Sola Martínez Adrián Martínez Vivancos Manuel Cánovas Díaz Teresa de Diego Puente |
| author_facet | Gema Lozano Terol Julia Gallego-Jara Rosa Alba Sola Martínez Adrián Martínez Vivancos Manuel Cánovas Díaz Teresa de Diego Puente |
| author_sort | Gema Lozano Terol |
| collection | DOAJ |
| description | Recombinant protein production for medical, academic, or industrial applications is essential for our current life. Recombinant proteins are obtained mainly through microbial fermentation, with Escherichia coli being the host most used. In spite of that, some problems are associated with the production of recombinant proteins in E. coli, such as the formation of inclusion bodies, the metabolic burden, or the inefficient translocation/transport system of expressed proteins. Optimizing transcription of heterologous genes is essential to avoid these drawbacks and develop competitive biotechnological processes. Here, expression of YFP reporter protein is evaluated under the control of four promoters of different strength (PT7lac, Ptrc, Ptac, and PBAD) and two different replication origins (high copy number pMB1′ and low copy number p15A). In addition, the study has been carried out with the E. coli BL21 wt and the ackA mutant strain growing in a rich medium with glucose or glycerol as carbon sources. Results showed that metabolic burden associated with transcription and translation of foreign genes involves a decrease in recombinant protein expression. It is necessary to find a balance between plasmid copy number and promoter strength to maximize soluble recombinant protein expression. The results obtained represent an important advance on the most suitable expression system to improve both the quantity and quality of recombinant proteins in bioproduction engineering. |
| first_indexed | 2024-12-21T10:05:14Z |
| format | Article |
| id | doaj.art-c339dd4234374b0097241b6407215ac4 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-21T10:05:14Z |
| publishDate | 2021-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-c339dd4234374b0097241b6407215ac42022-12-21T19:07:51ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2021-06-011210.3389/fmicb.2021.682001682001Impact of the Expression System on Recombinant Protein Production in Escherichia coli BL21Gema Lozano TerolJulia Gallego-JaraRosa Alba Sola MartínezAdrián Martínez VivancosManuel Cánovas DíazTeresa de Diego PuenteRecombinant protein production for medical, academic, or industrial applications is essential for our current life. Recombinant proteins are obtained mainly through microbial fermentation, with Escherichia coli being the host most used. In spite of that, some problems are associated with the production of recombinant proteins in E. coli, such as the formation of inclusion bodies, the metabolic burden, or the inefficient translocation/transport system of expressed proteins. Optimizing transcription of heterologous genes is essential to avoid these drawbacks and develop competitive biotechnological processes. Here, expression of YFP reporter protein is evaluated under the control of four promoters of different strength (PT7lac, Ptrc, Ptac, and PBAD) and two different replication origins (high copy number pMB1′ and low copy number p15A). In addition, the study has been carried out with the E. coli BL21 wt and the ackA mutant strain growing in a rich medium with glucose or glycerol as carbon sources. Results showed that metabolic burden associated with transcription and translation of foreign genes involves a decrease in recombinant protein expression. It is necessary to find a balance between plasmid copy number and promoter strength to maximize soluble recombinant protein expression. The results obtained represent an important advance on the most suitable expression system to improve both the quantity and quality of recombinant proteins in bioproduction engineering.https://www.frontiersin.org/articles/10.3389/fmicb.2021.682001/fullEscherichia colirecombinant proteinexpression systempromoterorigin of replicationmicrobial factory |
| spellingShingle | Gema Lozano Terol Julia Gallego-Jara Rosa Alba Sola Martínez Adrián Martínez Vivancos Manuel Cánovas Díaz Teresa de Diego Puente Impact of the Expression System on Recombinant Protein Production in Escherichia coli BL21 Frontiers in Microbiology Escherichia coli recombinant protein expression system promoter origin of replication microbial factory |
| title | Impact of the Expression System on Recombinant Protein Production in Escherichia coli BL21 |
| title_full | Impact of the Expression System on Recombinant Protein Production in Escherichia coli BL21 |
| title_fullStr | Impact of the Expression System on Recombinant Protein Production in Escherichia coli BL21 |
| title_full_unstemmed | Impact of the Expression System on Recombinant Protein Production in Escherichia coli BL21 |
| title_short | Impact of the Expression System on Recombinant Protein Production in Escherichia coli BL21 |
| title_sort | impact of the expression system on recombinant protein production in escherichia coli bl21 |
| topic | Escherichia coli recombinant protein expression system promoter origin of replication microbial factory |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2021.682001/full |
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