BioChem - DOS

Surface Enhanced Raman Scattering for Quantification of p-Coumaric Acid Produced by Escherichia coli

Analytical chemistry

Morelli, Lidia; Zó r, Kinga; Jendresen, Christian Bille; Rindzevicius, Tomas; Schmidt, Michael Stenbæk; Nielsen, Alex Toftgaard; Boisen, Anja

<P>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&ndash;liquid extraction (LLE) for quantification of <I>p</I>-coumaric acid (pHCA) in the supernatant of genetically engineered <I>Escherichia coli</I> (<I>E. coli</I>) cultures. pHCA was measured in a dynamic range from 1 &mu;M up to 50 &mu;M 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 <I>E. coli</I> 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.</P><P><B>Graphic Abstract</B><BR><IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/ancham/2017/ancham.2017.89.issue-7/acs.analchem.6b04428/production/images/medium/ac-2016-044283_0006.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/ac6b04428'>ACS Electronic Supporting Info</A></P>

2017

American Chemical Society

0003-2700

1520-6882

89

7

pp.3981-3987

10.1021/acs.analchem.6b04428

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART77682734