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Sensors 2016, 16(10), 1604; doi:10.3390/s16101604

A Toolbox of Genetically Encoded FRET-Based Biosensors for Rapid l-Lysine Analysis

1
IBG-1, Forschungszentrum Jülich GmbH, 52425 Jülich, Germany
2
ICS-6, Forschungszentrum Jülich GmbH, 52425 Jülich, Germany
*
Author to whom correspondence should be addressed.
Academic Editors: Niko Hildebrandt, Igor Medintz and Russ Algar
Received: 2 August 2016 / Revised: 15 September 2016 / Accepted: 20 September 2016 / Published: 28 September 2016
(This article belongs to the Special Issue FRET Biosensors)
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Abstract

Background: The fast development of microbial production strains for basic and fine chemicals is increasingly carried out in small scale cultivation systems to allow for higher throughput. Such parallelized systems create a need for new rapid online detection systems to quantify the respective target compound. In this regard, biosensors, especially genetically encoded Förster resonance energy transfer (FRET)-based biosensors, offer tremendous opportunities. As a proof-of-concept, we have created a toolbox of FRET-based biosensors for the ratiometric determination of l-lysine in fermentation broth. Methods: The sensor toolbox was constructed based on a sensor that consists of an optimized central lysine-/arginine-/ornithine-binding protein (LAO-BP) flanked by two fluorescent proteins (enhanced cyan fluorescent protein (ECFP), Citrine). Further sensor variants with altered affinity and sensitivity were obtained by circular permutation of the binding protein as well as the introduction of flexible and rigid linkers between the fluorescent proteins and the LAO-BP, respectively. Results: The sensor prototype was applied to monitor the extracellular l-lysine concentration of the l-lysine producing Corynebacterium glutamicum (C. glutamicum) strain DM1933 in a BioLector® microscale cultivation device. The results matched well with data obtained by HPLC analysis and the Ninhydrin assay, demonstrating the high potential of FRET-based biosensors for high-throughput microbial bioprocess optimization. View Full-Text
Keywords: FRET-based biosensors; l-lysine detection; metabolite analysis; online monitoring; microbial strain development FRET-based biosensors; l-lysine detection; metabolite analysis; online monitoring; microbial strain development
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Steffen, V.; Otten, J.; Engelmann, S.; Radek, A.; Limberg, M.; Koenig, B.W.; Noack, S.; Wiechert, W.; Pohl, M. A Toolbox of Genetically Encoded FRET-Based Biosensors for Rapid l-Lysine Analysis. Sensors 2016, 16, 1604.

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