Influence of Varying Pre-Culture Conditions on the Level of Population Heterogeneity in Batch Cultures with an <i>Escherichia coli</i> Triple Reporter Strain
When targeting robust, high-yielding bioprocesses, phenomena such as population heterogeneity have to be considered. Therefore, the influence of the conditions which the cells experience prior to the main culture should also be evaluated. Here, the influence of a pre-culture medium (complex vs. mini...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-07-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/11/7/1763 |
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| author | Manh Dat Hoang Sophi Riessner Jose Enrique Oropeza Vargas Nikolas von den Eichen Anna-Lena Heins |
| author_facet | Manh Dat Hoang Sophi Riessner Jose Enrique Oropeza Vargas Nikolas von den Eichen Anna-Lena Heins |
| author_sort | Manh Dat Hoang |
| collection | DOAJ |
| description | When targeting robust, high-yielding bioprocesses, phenomena such as population heterogeneity have to be considered. Therefore, the influence of the conditions which the cells experience prior to the main culture should also be evaluated. Here, the influence of a pre-culture medium (complex vs. minimal medium), optical density for inoculation of the main culture (0.005, 0.02 and 0.0125) and harvest time points of the pre-culture in exponential growth phase (early, mid and late) on the level of population heterogeneity in batch cultures of the <i>Escherichia coli</i> triple reporter strain G7<sub>BL21(DE3)</sub> in stirred-tank bioreactors was studied. This strain allows monitoring the growth (<i>rrnB</i>-EmGFP), general stress response (<i>rpoS</i>-mStrawberry) and oxygen limitation (<i>nar</i>-TagRFP657) of single cells through the expression of fluorescent proteins. Data from batch cultivations with varying pre-culture conditions were analysed with principal component analysis. According to fluorescence data, the pre-culture medium had the largest impact on population heterogeneities during the bioprocess. While a minimal medium as a pre-culture medium elevated the differences in cellular growth behaviour in the subsequent batch process, a complex medium increased the general stress response and led to a higher population heterogeneity. The latter was promoted by an early harvest of the cells with low inoculation density. Seemingly, <i>nar</i>-operon expression acted independently of the pre-culture conditions. |
| first_indexed | 2024-03-11T00:49:06Z |
| format | Article |
| id | doaj.art-a80a71cb43f348b99e35dfc965962330 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-11T00:49:06Z |
| publishDate | 2023-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-a80a71cb43f348b99e35dfc9659623302023-11-18T20:36:07ZengMDPI AGMicroorganisms2076-26072023-07-01117176310.3390/microorganisms11071763Influence of Varying Pre-Culture Conditions on the Level of Population Heterogeneity in Batch Cultures with an <i>Escherichia coli</i> Triple Reporter StrainManh Dat Hoang0Sophi Riessner1Jose Enrique Oropeza Vargas2Nikolas von den Eichen3Anna-Lena Heins4Chair of Biochemical Engineering, TUM School of Engineering and Design, Technical University of Munich, 85748 Garching, GermanyChair of Biochemical Engineering, TUM School of Engineering and Design, Technical University of Munich, 85748 Garching, GermanyChair of Biochemical Engineering, TUM School of Engineering and Design, Technical University of Munich, 85748 Garching, GermanyChair of Biochemical Engineering, TUM School of Engineering and Design, Technical University of Munich, 85748 Garching, GermanyChair of Biochemical Engineering, TUM School of Engineering and Design, Technical University of Munich, 85748 Garching, GermanyWhen targeting robust, high-yielding bioprocesses, phenomena such as population heterogeneity have to be considered. Therefore, the influence of the conditions which the cells experience prior to the main culture should also be evaluated. Here, the influence of a pre-culture medium (complex vs. minimal medium), optical density for inoculation of the main culture (0.005, 0.02 and 0.0125) and harvest time points of the pre-culture in exponential growth phase (early, mid and late) on the level of population heterogeneity in batch cultures of the <i>Escherichia coli</i> triple reporter strain G7<sub>BL21(DE3)</sub> in stirred-tank bioreactors was studied. This strain allows monitoring the growth (<i>rrnB</i>-EmGFP), general stress response (<i>rpoS</i>-mStrawberry) and oxygen limitation (<i>nar</i>-TagRFP657) of single cells through the expression of fluorescent proteins. Data from batch cultivations with varying pre-culture conditions were analysed with principal component analysis. According to fluorescence data, the pre-culture medium had the largest impact on population heterogeneities during the bioprocess. While a minimal medium as a pre-culture medium elevated the differences in cellular growth behaviour in the subsequent batch process, a complex medium increased the general stress response and led to a higher population heterogeneity. The latter was promoted by an early harvest of the cells with low inoculation density. Seemingly, <i>nar</i>-operon expression acted independently of the pre-culture conditions.https://www.mdpi.com/2076-2607/11/7/1763pre-culturepopulation heterogeneitytriple reporter strainbatch cultureprincipal component analysis |
| spellingShingle | Manh Dat Hoang Sophi Riessner Jose Enrique Oropeza Vargas Nikolas von den Eichen Anna-Lena Heins Influence of Varying Pre-Culture Conditions on the Level of Population Heterogeneity in Batch Cultures with an <i>Escherichia coli</i> Triple Reporter Strain Microorganisms pre-culture population heterogeneity triple reporter strain batch culture principal component analysis |
| title | Influence of Varying Pre-Culture Conditions on the Level of Population Heterogeneity in Batch Cultures with an <i>Escherichia coli</i> Triple Reporter Strain |
| title_full | Influence of Varying Pre-Culture Conditions on the Level of Population Heterogeneity in Batch Cultures with an <i>Escherichia coli</i> Triple Reporter Strain |
| title_fullStr | Influence of Varying Pre-Culture Conditions on the Level of Population Heterogeneity in Batch Cultures with an <i>Escherichia coli</i> Triple Reporter Strain |
| title_full_unstemmed | Influence of Varying Pre-Culture Conditions on the Level of Population Heterogeneity in Batch Cultures with an <i>Escherichia coli</i> Triple Reporter Strain |
| title_short | Influence of Varying Pre-Culture Conditions on the Level of Population Heterogeneity in Batch Cultures with an <i>Escherichia coli</i> Triple Reporter Strain |
| title_sort | influence of varying pre culture conditions on the level of population heterogeneity in batch cultures with an i escherichia coli i triple reporter strain |
| topic | pre-culture population heterogeneity triple reporter strain batch culture principal component analysis |
| url | https://www.mdpi.com/2076-2607/11/7/1763 |
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