Journal article
Surface Enhanced Raman Scattering for Quantification of p-Coumaric Acid Produced by Escherichia coli
Department of Micro- and Nanotechnology, Technical University of Denmark1
Nanoprobes, Department of Micro- and Nanotechnology, Technical University of Denmark2
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
Bacterial Cell Factory Optimization, Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark4
Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark5
Center for Intelligent Drug Delivery and Sensing Using Microcontainers and Nanomechanics, Department of Health Technology, Technical University of Denmark6
The number of newly developed genetic variants of microbial cell factories for production of biochemicals has been rapidly growing in recent years, leading to an increased need for new screening techniques. We developed a method based on surface-enhanced Raman scattering (SERS) coupled with liquid-liquid extraction (LLE) for quantification of p-coumaric acid (pHCA) in the supernatant of genetically engineered Escherichia coli (E. coli) cultures. pHCA was measured in a dynamic range from 1 μM up to 50 μM on highly uniform SERS substrates based on leaning gold-capped nanopillars, which showed an in-wafer signal variation of only 11.7%.
LLE using dichloromethane as organic phase was combined with the detection in order to increase selectivity and sensitivity by decreasing the effect of interfering compounds from the analytes of interest. The difference in pHCA production yield between three genetically engineered E. coli strains was successfully evaluated using SERS and confirmed with high-performance liquid chromatography.
As this novel approach has potential to be automated and parallelized, it can be considered for high-throughput screening in metabolic engineering.
Language: | English |
---|---|
Publisher: | American Chemical Society (ACS) |
Year: | 2017 |
Pages: | 3981-3987 |
ISSN: | 15206882 , 15204782 and 00032700 |
Types: | Journal article |
DOI: | 10.1021/acs.analchem.6b04428 |
ORCIDs: | Zor, Kinga , Jendresen, Christian Bille , Rindzevicius, Tomas , Nielsen, Alex Toftgaard , Boisen, Anja and 0000-0002-3299-8608 |