The application of two-dimensional fluorescence spectroscopy for the on-line evaluation of modified enzymatic enantioselectivities in organic solvents by forming substrate salts

Abstract We recently developed a pseudo-enantiomeric reaction which was monitored on-line via 2D-fluorescence spectroscopy in different reaction media. During enzymatic reactions, conclusions about the converted substrates and the enantioselectivity could be drawn immediately, using two different fluorescence spectroscopic detectable substrates, l -phenylalanine-7-amido-4-methylcoumarine and d -phenylalanine-7-amido-4-trifluoromethylcoumarine. Now, due to the enlargement of one of the coumarin substrates via salt formation, it was possible to influence the enzymatyic enantioselectivity in toluene and to monitor the impact of the bulky counterion on-line.

[1]  P. Das,et al.  Binding of hydrophobic hydroxamic acids enhances peroxidase's stereoselectivity in nonaqueous sulfoxidations. , 2002, Journal of the American Chemical Society.

[2]  E. Vulfson,et al.  At what temperature can enzymes maintain their catalytic activity? , 2000, Enzyme and microbial technology.

[3]  Thomas Scheper,et al.  On-line monitoring of a quasi-enantiomeric reaction with two coumarin substrates via 2D-fluorescence spectroscopy , 2001 .

[4]  T. Scheper,et al.  Synthesis, test and application of chirale fluorescence substrates to evaluate enzymatic processes in different reaction media , 2005 .

[5]  Per Berglund,et al.  Engineered enzymes for improved organic synthesis. , 2003, Current opinion in biotechnology.

[6]  J. Dordick,et al.  Enzyme activation for nonaqueous media. , 2002, Current opinion in biotechnology.

[7]  A. Klibanov,et al.  Improving lipase enantioselectivity in organic solvents by forming substrate salts with chiral agents. , 2000, Biotechnology and bioengineering.

[8]  P. Halling Biocatalysis in low-water media: understanding effects of reaction conditions. , 2000, Current opinion in chemical biology.

[9]  A. Klibanov Asymmetric enzymatic oxidoreductions in organic solvents. , 2003, Current opinion in biotechnology.

[10]  C. Sih,et al.  Quantitative analyses of biochemical kinetic resolutions of enantiomers , 1982 .

[11]  A. Klibanov,et al.  MARKEDLY ENHANCING ENZYMATIC ENANTIOSELECTIVITY IN ORGANIC SOLVENTS BY FORMING SUBSTRATE SALTS , 1999 .