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| 024 | 7 | _ | |a 10.1186/s12934-019-1193-y |2 doi |
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| 100 | 1 | _ | |a Otten, Julia |0 P:(DE-Juel1)165764 |b 0 |
| 245 | _ | _ | |a A FRET-based biosensor for the quantification of glucose in culture supernatants of mL scale microbial cultivations |
| 260 | _ | _ | |a London |c 2019 |b Biomed Central |
| 336 | 7 | _ | |a article |2 DRIVER |
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| 520 | _ | _ | |a BackgroundIn most microbial cultivations d-glucose is the main carbon and energy source. However, quantification of d-glucose especially in small scale is still challenging. Therefore, we developed a FRET-based glucose biosensor, which can be applied in microbioreactor-based cultivations. This sensor consists of a glucose binding protein sandwiched between two fluorescent proteins, constituting a FRET pair. Upon d-glucose binding the sensor undergoes a conformational change which is translated into a FRET-ratio change.ResultsThe selected sensor shows an apparent Kd below 1.5 mM d-glucose and a very high sensitivity of up to 70% FRET-ratio change between the unbound and the glucose-saturated state. The soluble sensor was successfully applied online to monitor the glucose concentration in an Escherichia coli culture. Additionally, this sensor was utilized in an at-line process for a Corynebacterium glutamicum culture as an example for a process with cell-specific background (e.g. autofluorescence) and medium-induced quenching. Immobilization of the sensor via HaloTag® enabled purification and covalent immobilization in one step and increased the stability during application, significantly.ConclusionA FRET-based glucose sensor was used to quantify d-glucose consumption in microtiter plate based cultivations. To the best of our knowledge, this is the first method reported for online quantification of d-glucose in microtiter plate based cultivations. In comparison to d-glucose analysis via an enzymatic assay and HPLC, the sensor performed equally well, but enabled much faster measurements, which allowed to speed up microbial strain development significantly. |
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| 700 | 1 | _ | |a Tenhaef, Niklas |0 P:(DE-Juel1)168172 |b 1 |
| 700 | 1 | _ | |a Jansen, Roman P. |0 P:(DE-Juel1)171232 |b 2 |
| 700 | 1 | _ | |a Döbber, Johannes |0 P:(DE-Juel1)166538 |b 3 |
| 700 | 1 | _ | |a Jungbluth, Lisa |0 P:(DE-Juel1)177977 |b 4 |
| 700 | 1 | _ | |a Noack, Stephan |0 P:(DE-Juel1)129050 |b 5 |
| 700 | 1 | _ | |a Oldiges, Marco |0 P:(DE-Juel1)129053 |b 6 |
| 700 | 1 | _ | |a Wiechert, Wolfgang |0 P:(DE-Juel1)129076 |b 7 |
| 700 | 1 | _ | |a Pohl, Martina |0 P:(DE-Juel1)131522 |b 8 |e Corresponding author |
| 773 | _ | _ | |a 10.1186/s12934-019-1193-y |g Vol. 18, no. 1, p. 143 |0 PERI:(DE-600)2091377-1 |n 1 |p 143 |t Microbial cell factories |v 18 |y 2019 |x 1475-2859 |
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