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| Content Provider | Springer Nature : BioMed Central |
|---|---|
| Author | Unthan, Simon Radek, Andreas Wiechert, Wolfgang Oldiges, Marco Noack, Stephan |
| Abstract | Background The throughput of cultivation experiments in bioprocess development has drastically increased in recent years due to the availability of sophisticated microliter scale cultivation devices. However, as these devices still require time-consuming manual work, the bottleneck was merely shifted to media preparation, inoculation and finally the analyses of cultivation samples. A first step towards solving these issues was undertaken in our former study by embedding a BioLector in a robotic workstation. This workstation already allowed for the optimization of heterologous protein production processes, but remained limited when aiming for the characterization of small molecule producer strains. In this work, we extended our workstation to a versatile Mini Pilot Plant (MPP) by integrating further robotic workflows and microtiter plate assays that now enable a fast and accurate phenotyping of a broad range of microbial production hosts. Results A fully automated harvest procedure was established, which repeatedly samples up to 48 wells from BioLector cultivations in response to individually defined trigger conditions. The samples are automatically clarified by centrifugation and finally frozen for subsequent analyses. Sensitive metabolite assays in 384-well plate scale were integrated on the MPP for the direct determination of substrate uptake (specifically D-glucose and D-xylose) and product formation (specifically amino acids). In a first application, we characterized a set of Corynebacterium glutamicum L-lysine producer strains and could rapidly identify a unique strain showing increased L-lysine titers, which was subsequently confirmed in lab-scale bioreactor experiments. In a second study, we analyzed the substrate uptake kinetics of a previously constructed D-xylose-converting C. glutamicum strain during cultivation on mixed carbon sources in a fully automated experiment. Conclusions The presented MPP is designed to face the challenges typically encountered during early-stage bioprocess development. Especially the bottleneck of sample analyses from fast and parallelized microtiter plate cultivations can be solved using cutting-edge robotic automation. As robotic workstations become increasingly attractive for biotechnological research, we expect our setup to become a template for future bioprocess development. |
| Related Links | https://microbialcellfactories.biomedcentral.com/counter/pdf/10.1186/s12934-015-0216-6.pdf |
| Ending Page | 11 |
| Page Count | 11 |
| Starting Page | 1 |
| File Format | HTM / HTML |
| ISSN | 14752859 |
| DOI | 10.1186/s12934-015-0216-6 |
| Journal | Microbial Cell Factories |
| Issue Number | 1 |
| Volume Number | 14 |
| Language | English |
| Publisher | BioMed Central |
| Publisher Date | 2015-03-11 |
| Access Restriction | Open |
| Subject Keyword | Applied Microbiology Biotechnology Microbiology Microbial Genetics and Genomics Enzymology Genetic Engineering C. glutamicum Mini Pilot Plant Bioprocess automation Microtiter plate assay Scale-up Scale-down High-throughput Screening Microbial phenotyping D-xylose Ninhydrin L-lysine |
| Content Type | Text |
| Resource Type | Article |
| Subject | Applied Microbiology and Biotechnology Bioengineering Biotechnology |
| Journal Impact Factor | 4.3/2023 |
| 5-Year Journal Impact Factor | 5.5/2023 |
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