Optimized polymer-based glucose release in microtiter plates for small-scale E. coli fed-batch cultivations
dc.bibliographicCitation.firstPage | 24 | eng |
dc.bibliographicCitation.issue | 1 | eng |
dc.bibliographicCitation.journalTitle | Journal of Biological Engineering | eng |
dc.bibliographicCitation.volume | 14 | eng |
dc.contributor.author | Keil, Timm | |
dc.contributor.author | Dittrich, Barbara | |
dc.contributor.author | Lattermann, Clemens | |
dc.contributor.author | Büchs, Jochen | |
dc.date.accessioned | 2021-07-30T08:56:05Z | |
dc.date.available | 2021-07-30T08:56:05Z | |
dc.date.issued | 2020 | |
dc.description.abstract | Background: Small-scale cultivation vessels, which allow fed-batch operation mode, become more and more important for fast and reliable early process development. Recently, the polymer-based feeding system was introduced to allow fed-batch conditions in microtiter plates. Maximum glucose release rates of 0.35 mg/h per well (48-well-plate) at 37 °C can be achieved with these plates, depending on the media properties. The fed-batch cultivation of fluorescent protein-expressing E. coli at oxygen transfer rate levels of 5 mmol/L/h proved to be superior compared to simple batch cultivations. However, literature suggests that higher glucose release rates than achieved with the currently available fed-batch microtiter plate are beneficial, especially for fast-growing microorganisms. During the fed-batch phase of the cultivation, a resulting oxygen transfer rate level of 28 mmol/L/h should be achieved. Results: Customization of the polymer matrix enabled a considerable increase in the glucose release rate of more than 250% to up to 0.90 mg/h per well. Therefore, the molecular weight of the prepolymer and the addition of a hydrophilic PDMS-PEG copolymer allowed for the individual adjustment of a targeted glucose release rate. The newly developed polymer matrix was additionally invariant to medium properties like the osmotic concentration or the pH-value. The glucose release rate of the optimized matrix was constant in various synthetic and complex media. Fed-batch cultivations of E. coli in microtiter plates with the optimized matrix revealed elevated oxygen transfer rates during the fed-batch phase of approximately 28 mmol/L/h. However, these increased glucose release rates resulted in a prolonged initial batch phase and oxygen limitations. The newly developed polymer-based feeding system provides options to manufacture individual feed rates in a range from 0.24-0.90 mg/h per well. Conclusions: The optimized polymer-based fed-batch microtiter plate allows higher reproducibility of fed-batch experiments since cultivation media properties have almost no influence on the release rate. The adjustment of individual feeding rates in a wide range supports the early process development for slow, average and fast-growing microorganisms in microtiter plates. The study underlines the importance of a detailed understanding of the metabolic behavior (through online monitoring techniques) to identify optimal feed rates. © 2020 The Author(s). | eng |
dc.description.version | publishedVersion | eng |
dc.identifier.uri | https://oa.tib.eu/renate/handle/123456789/6462 | |
dc.identifier.uri | https://doi.org/10.34657/5509 | |
dc.language.iso | eng | eng |
dc.publisher | London : BioMed Central | eng |
dc.relation.doi | https://doi.org/10.1186/s13036-020-00247-0 | |
dc.relation.essn | 1754-1611 | |
dc.rights.license | CC BY 4.0 Unported | eng |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | eng |
dc.subject.ddc | 570 | eng |
dc.subject.other | Bioprocess development | eng |
dc.subject.other | Catabolite repression | eng |
dc.subject.other | Escherichia coli | eng |
dc.subject.other | Fed-batch | eng |
dc.subject.other | High-throughput | eng |
dc.subject.other | Microtiter plate | eng |
dc.subject.other | Overflow metabolism | eng |
dc.title | Optimized polymer-based glucose release in microtiter plates for small-scale E. coli fed-batch cultivations | eng |
dc.type | Article | eng |
dc.type | Text | eng |
tib.accessRights | openAccess | eng |
wgl.contributor | DWI | eng |
wgl.subject | Biowissensschaften/Biologie | eng |
wgl.type | Zeitschriftenartikel | eng |
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