Improving E. coli growth performance by manipulating small RNA expression
Abstract Efficient growth of E. coli, especially for production of recombinant proteins, has been a challenge for the biotechnological industry since the early 1970s. By employing multiple approaches, such as different media composition, various growth strategies and specific genetic manipulations,...
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| Format: | Article |
| Language: | English |
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BMC
2017-11-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12934-017-0810-x |
| _version_ | 1828378397882974208 |
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| author | Alejandro Negrete Joseph Shiloach |
| author_facet | Alejandro Negrete Joseph Shiloach |
| author_sort | Alejandro Negrete |
| collection | DOAJ |
| description | Abstract Efficient growth of E. coli, especially for production of recombinant proteins, has been a challenge for the biotechnological industry since the early 1970s. By employing multiple approaches, such as different media composition, various growth strategies and specific genetic manipulations, it is now possible to grow bacteria to concentrations exceeding 100 g/L and to achieve high concentrations of recombinant proteins. Although the growth conditions are carefully monitored and maintained, it is likely that during the growth process cells are exposed to periodic stress conditions, created by fluctuations in pH, dissolved oxygen, temperature, glucose, and salt concentration. These stress circumstances which can occur especially in large volume bioreactors, may affect the growth and production process. In the last several years, it has been recognized that small non-coding RNAs can act as regulators of bacterial gene expression. These molecules are found to be specifically involved in E. coli response to different environmental stress conditions; but so far, have not been used for improving production strains. The review provides summary of small RNAs identified on petri dish or in shake flask culture that can potentially affect growth characteristics of E. coli grown in bioreactor. Among them MicC and MicF that are involved in response to temperature changes, RyhB that responds to iron concentration, Gady which is associated with lower pH, Sgrs that is coupled with glucose transport and OxyS that responds to oxygen concentration. The manipulation of some of these small RNAs for improving growth of E. coli in Bioreactor is described in the last part of the review. Overexpression of SgrS was associated with improved growth and reduced acetate expression, over expression of GadY improved cell growth at acidic conditions and over expression of OxyS reduced the effect of oxidative stress. One of the possible advantages of manipulating sRNAs for improving cell growth is that the modifications occur at a post-translational level. Therefore, the use of sRNAs may exert minimal effect on the overall bacterial metabolism. The elucidation of the physiological role of newly discovered sRNAs will open new possibilities for creating strains with improved growth and production capabilities. |
| first_indexed | 2024-04-14T08:26:23Z |
| format | Article |
| id | doaj.art-aeeb76f6596f4ff6a5137f62b2d041fc |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-14T08:26:23Z |
| publishDate | 2017-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-aeeb76f6596f4ff6a5137f62b2d041fc2022-12-22T02:04:03ZengBMCMicrobial Cell Factories1475-28592017-11-0116111210.1186/s12934-017-0810-xImproving E. coli growth performance by manipulating small RNA expressionAlejandro Negrete0Joseph Shiloach1Biotechnology Core Laboratory, NIDDK, NIHBiotechnology Core Laboratory, NIDDK, NIHAbstract Efficient growth of E. coli, especially for production of recombinant proteins, has been a challenge for the biotechnological industry since the early 1970s. By employing multiple approaches, such as different media composition, various growth strategies and specific genetic manipulations, it is now possible to grow bacteria to concentrations exceeding 100 g/L and to achieve high concentrations of recombinant proteins. Although the growth conditions are carefully monitored and maintained, it is likely that during the growth process cells are exposed to periodic stress conditions, created by fluctuations in pH, dissolved oxygen, temperature, glucose, and salt concentration. These stress circumstances which can occur especially in large volume bioreactors, may affect the growth and production process. In the last several years, it has been recognized that small non-coding RNAs can act as regulators of bacterial gene expression. These molecules are found to be specifically involved in E. coli response to different environmental stress conditions; but so far, have not been used for improving production strains. The review provides summary of small RNAs identified on petri dish or in shake flask culture that can potentially affect growth characteristics of E. coli grown in bioreactor. Among them MicC and MicF that are involved in response to temperature changes, RyhB that responds to iron concentration, Gady which is associated with lower pH, Sgrs that is coupled with glucose transport and OxyS that responds to oxygen concentration. The manipulation of some of these small RNAs for improving growth of E. coli in Bioreactor is described in the last part of the review. Overexpression of SgrS was associated with improved growth and reduced acetate expression, over expression of GadY improved cell growth at acidic conditions and over expression of OxyS reduced the effect of oxidative stress. One of the possible advantages of manipulating sRNAs for improving cell growth is that the modifications occur at a post-translational level. Therefore, the use of sRNAs may exert minimal effect on the overall bacterial metabolism. The elucidation of the physiological role of newly discovered sRNAs will open new possibilities for creating strains with improved growth and production capabilities.http://link.springer.com/article/10.1186/s12934-017-0810-xSmall RNAStress responseHigh cell density culturesE. coliAcetate production |
| spellingShingle | Alejandro Negrete Joseph Shiloach Improving E. coli growth performance by manipulating small RNA expression Microbial Cell Factories Small RNA Stress response High cell density cultures E. coli Acetate production |
| title | Improving E. coli growth performance by manipulating small RNA expression |
| title_full | Improving E. coli growth performance by manipulating small RNA expression |
| title_fullStr | Improving E. coli growth performance by manipulating small RNA expression |
| title_full_unstemmed | Improving E. coli growth performance by manipulating small RNA expression |
| title_short | Improving E. coli growth performance by manipulating small RNA expression |
| title_sort | improving e coli growth performance by manipulating small rna expression |
| topic | Small RNA Stress response High cell density cultures E. coli Acetate production |
| url | http://link.springer.com/article/10.1186/s12934-017-0810-x |
| work_keys_str_mv | AT alejandronegrete improvingecoligrowthperformancebymanipulatingsmallrnaexpression AT josephshiloach improvingecoligrowthperformancebymanipulatingsmallrnaexpression |