Journal Article

FZJ-2016-06487

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png A Toolbox of Genetically Encoded FRET-Based Biosensors for Rapid l-Lysine Analysis

Steffen, V.FZJ* ; Otten, J.FZJ* ; Engelmann, S. ; Radek, A.FZJ* ; Limberg, M.FZJ* ; König, B.FZJ* ; Noack, S.FZJ* ; Wiechert, W.FZJ* ; Pohl, M. (Corresponding author)FZJ*

2016
MDPI Basel

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Please use a persistent id in citations: http://hdl.handle.net/2128/12927  doi:10.3390/s16101604

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.

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Contributing Institute(s):

1. Strukturbiochemie (ICS-6)
2. Biotechnologie (IBG-1)

Research Program(s):

1. 553 - Physical Basis of Diseases (POF3-553) (POF3-553)

Appears in the scientific report 2016

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Database coverage:
; ; ; ; DOAJ Seal ; Ebsco Academic Search ; IF < 5 ; JCR ; NCBI Molecular Biology Database ; SCOPUS ; Science Citation Index Expanded ; Thomson Reuters Master Journal List ; Web of Science Core Collection

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