On-line monitoring of a quasi-enantiomeric reaction with two coumarin substrates via 2D-fluorescence spectroscopy

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OriginalspracheEnglisch
Seiten (von - bis)150-159
Seitenumfang10
FachzeitschriftEnzyme and microbial technology
Jahrgang29
Ausgabenummer2-3
PublikationsstatusVeröffentlicht - 17 Juli 2001

Abstract

With the use of two fluorescence spectroscopic detectable substrates, L-phenylalanin-7-amido-4-methylcoumarin (L-PheAMC) and D-phenylalanin-7-amido-4-trifluoromethylcoumarin (D-PheAFC), it was possible to monitor a quasi-enantiomeric enzymatic reaction. The simultaneous hydrolysis of L-PheAMC and D-PheAFC was applied in aqueous media and via on-line 2D-fluorescence spectroscopy it was possible to draw conclusions about the enantioselectivity of the used enzymes α-chymotrypsin and esterase from porcine liver. The kinetic parameters from the single-hydrolysis of L-PheAMC and D-PheAFC were determined via a timescan run, the results showed, that the L-substrate was hydrolyzed 106 times faster than the D-substrate by α-chymotrypsin. In a simultaneous usage of both coumarin substrates with α-chymotrypsin, the L-coumarin was favored by the enzyme, even when the L-substrate was used in a 10 times lower concentration than the D-substrate. With the unspecific esterase, the L-substrate was first hydrolyzed again when equal concentrations of both coumarins were used. But the reaction was influenced by the presence of the D-substrate. With a 10 times lower L-substrate concentration, the D-substrate was hydrolyzed by the esterase, but the reaction was again influenced by the presence of the enantiomeric partner.

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On-line monitoring of a quasi-enantiomeric reaction with two coumarin substrates via 2D-fluorescence spectroscopy. / Knüttel, Torsten; Hartmann, Thorsten; Meyer, Hartmut et al.
in: Enzyme and microbial technology, Jahrgang 29, Nr. 2-3, 17.07.2001, S. 150-159.

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Knüttel, Torsten ; Hartmann, Thorsten ; Meyer, Hartmut et al. / On-line monitoring of a quasi-enantiomeric reaction with two coumarin substrates via 2D-fluorescence spectroscopy. in: Enzyme and microbial technology. 2001 ; Jahrgang 29, Nr. 2-3. S. 150-159.
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abstract = "With the use of two fluorescence spectroscopic detectable substrates, L-phenylalanin-7-amido-4-methylcoumarin (L-PheAMC) and D-phenylalanin-7-amido-4-trifluoromethylcoumarin (D-PheAFC), it was possible to monitor a quasi-enantiomeric enzymatic reaction. The simultaneous hydrolysis of L-PheAMC and D-PheAFC was applied in aqueous media and via on-line 2D-fluorescence spectroscopy it was possible to draw conclusions about the enantioselectivity of the used enzymes α-chymotrypsin and esterase from porcine liver. The kinetic parameters from the single-hydrolysis of L-PheAMC and D-PheAFC were determined via a timescan run, the results showed, that the L-substrate was hydrolyzed 106 times faster than the D-substrate by α-chymotrypsin. In a simultaneous usage of both coumarin substrates with α-chymotrypsin, the L-coumarin was favored by the enzyme, even when the L-substrate was used in a 10 times lower concentration than the D-substrate. With the unspecific esterase, the L-substrate was first hydrolyzed again when equal concentrations of both coumarins were used. But the reaction was influenced by the presence of the D-substrate. With a 10 times lower L-substrate concentration, the D-substrate was hydrolyzed by the esterase, but the reaction was again influenced by the presence of the enantiomeric partner.",
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author = "Torsten Kn{\"u}ttel and Thorsten Hartmann and Hartmut Meyer and Thomas Scheper",
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TY - JOUR

T1 - On-line monitoring of a quasi-enantiomeric reaction with two coumarin substrates via 2D-fluorescence spectroscopy

AU - Knüttel, Torsten

AU - Hartmann, Thorsten

AU - Meyer, Hartmut

AU - Scheper, Thomas

N1 - Funding information: The authors thank the Deutsche Forschungsgemeinschaft (DFG, as part of the Graduiertenkolleg) for financial support.

PY - 2001/7/17

Y1 - 2001/7/17

N2 - With the use of two fluorescence spectroscopic detectable substrates, L-phenylalanin-7-amido-4-methylcoumarin (L-PheAMC) and D-phenylalanin-7-amido-4-trifluoromethylcoumarin (D-PheAFC), it was possible to monitor a quasi-enantiomeric enzymatic reaction. The simultaneous hydrolysis of L-PheAMC and D-PheAFC was applied in aqueous media and via on-line 2D-fluorescence spectroscopy it was possible to draw conclusions about the enantioselectivity of the used enzymes α-chymotrypsin and esterase from porcine liver. The kinetic parameters from the single-hydrolysis of L-PheAMC and D-PheAFC were determined via a timescan run, the results showed, that the L-substrate was hydrolyzed 106 times faster than the D-substrate by α-chymotrypsin. In a simultaneous usage of both coumarin substrates with α-chymotrypsin, the L-coumarin was favored by the enzyme, even when the L-substrate was used in a 10 times lower concentration than the D-substrate. With the unspecific esterase, the L-substrate was first hydrolyzed again when equal concentrations of both coumarins were used. But the reaction was influenced by the presence of the D-substrate. With a 10 times lower L-substrate concentration, the D-substrate was hydrolyzed by the esterase, but the reaction was again influenced by the presence of the enantiomeric partner.

AB - With the use of two fluorescence spectroscopic detectable substrates, L-phenylalanin-7-amido-4-methylcoumarin (L-PheAMC) and D-phenylalanin-7-amido-4-trifluoromethylcoumarin (D-PheAFC), it was possible to monitor a quasi-enantiomeric enzymatic reaction. The simultaneous hydrolysis of L-PheAMC and D-PheAFC was applied in aqueous media and via on-line 2D-fluorescence spectroscopy it was possible to draw conclusions about the enantioselectivity of the used enzymes α-chymotrypsin and esterase from porcine liver. The kinetic parameters from the single-hydrolysis of L-PheAMC and D-PheAFC were determined via a timescan run, the results showed, that the L-substrate was hydrolyzed 106 times faster than the D-substrate by α-chymotrypsin. In a simultaneous usage of both coumarin substrates with α-chymotrypsin, the L-coumarin was favored by the enzyme, even when the L-substrate was used in a 10 times lower concentration than the D-substrate. With the unspecific esterase, the L-substrate was first hydrolyzed again when equal concentrations of both coumarins were used. But the reaction was influenced by the presence of the D-substrate. With a 10 times lower L-substrate concentration, the D-substrate was hydrolyzed by the esterase, but the reaction was again influenced by the presence of the enantiomeric partner.

KW - 2D-fluorescence

KW - Chymotrypsin

KW - Coumarin

KW - Esterase

KW - On-line-monitoring

UR - http://www.scopus.com/inward/record.url?scp=0035822718&partnerID=8YFLogxK

U2 - 10.1016/S0141-0229(01)00369-6

DO - 10.1016/S0141-0229(01)00369-6

M3 - Article

AN - SCOPUS:0035822718

VL - 29

SP - 150

EP - 159

JO - Enzyme and microbial technology

JF - Enzyme and microbial technology

SN - 0141-0229

IS - 2-3

ER -

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