TY - JOUR

T1 - Fluorescence spectroscopy as a novel method for on-line analysis of biocatalytic C-C bond formations

AU - Kara, Selin

AU - Anton, Fabienne

AU - Solle, Dörte

AU - Neumann, Markus

AU - Hitzmann, Bernd

AU - Scheper, Thomas

AU - Liese, Andreas

PY - 2010/4/16

Y1 - 2010/4/16

N2 - On-line analysis of bioprocesses is of increasing interest avoiding the time delay for off-line sample preparation and the following analyses via chromatographic methods. Moreover, continuous monitoring of the reaction components during chemo- or biocatalytic transformations provides a direct control of the process. Since productivity of the processes can be controlled simultaneously, on-line monitoring of the processes is attractive for industrial applications. The reliable in situ monitoring of biocatalyzed reactions has been a challenge where reactions run in aqueous solutions. Limited work has been published on the use of spectroscopic methods for on-line analysis of biocatalytic reactions up to now. However, in this communication two dimensional (2D)-fluorescence spectroscopy has been proved to be an effective tool for on-line monitoring of the carboligation reactions catalyzed by wild type benzoylformate decarboxylase (BFD) from Pseudomonas putida. BFD is a thiamine diphosphate (ThDP)-dependent enzyme that catalyzes the asymmetric C-C bond formation to (S)-2-hydroxypropiophenone ((S)-2-HPP) starting from benzaldehyde and acetaldehyde. The analysis of the fluorescence spectra was achieved by chemometric modeling performing principle component analysis (PCA) and partial least square (PLS) regression. The derived chemometric models were used for the validation of concentrations of yielded 2-HPP and the substrate benzaldehyde with low root mean square error of calibration (RMSEC).

AB - On-line analysis of bioprocesses is of increasing interest avoiding the time delay for off-line sample preparation and the following analyses via chromatographic methods. Moreover, continuous monitoring of the reaction components during chemo- or biocatalytic transformations provides a direct control of the process. Since productivity of the processes can be controlled simultaneously, on-line monitoring of the processes is attractive for industrial applications. The reliable in situ monitoring of biocatalyzed reactions has been a challenge where reactions run in aqueous solutions. Limited work has been published on the use of spectroscopic methods for on-line analysis of biocatalytic reactions up to now. However, in this communication two dimensional (2D)-fluorescence spectroscopy has been proved to be an effective tool for on-line monitoring of the carboligation reactions catalyzed by wild type benzoylformate decarboxylase (BFD) from Pseudomonas putida. BFD is a thiamine diphosphate (ThDP)-dependent enzyme that catalyzes the asymmetric C-C bond formation to (S)-2-hydroxypropiophenone ((S)-2-HPP) starting from benzaldehyde and acetaldehyde. The analysis of the fluorescence spectra was achieved by chemometric modeling performing principle component analysis (PCA) and partial least square (PLS) regression. The derived chemometric models were used for the validation of concentrations of yielded 2-HPP and the substrate benzaldehyde with low root mean square error of calibration (RMSEC).

KW - Benzoylformate decarboxylase

KW - C-C bond formation

KW - Chemometrics

KW - Enzymatic catalysis

KW - Fluorescence spectroscopy

KW - On-line monitoring

UR - http://www.scopus.com/inward/record.url?scp=77957827905&partnerID=8YFLogxK

U2 - 10.1016/j.molcatb.2010.04.006

DO - 10.1016/j.molcatb.2010.04.006

M3 - Article

AN - SCOPUS:77957827905

VL - 66

SP - 124

EP - 129

JO - Journal of Molecular Catalysis B: Enzymatic

JF - Journal of Molecular Catalysis B: Enzymatic

SN - 1381-1177

IS - 1-2

ER -