Details
| Original language | English |
|---|---|
| Pages (from-to) | 673-686 |
| Number of pages | 14 |
| Journal | Enzyme and microbial technology |
| Volume | 37 |
| Issue number | 7 |
| Publication status | Published - 14 Jun 2005 |
Abstract
A pseudo-enantiomeric reaction was developed which was monitored on-line via 2D-fluorescence spectroscopy. During enzymatic reactions the enantiomeric excess can be followed on-line. Fluorescence spectroscopic detectable substrates, l-/d-phenylalanine-7-amido-4-methylcoumarine (l-/d-PheAMC) and l-/d-phenylalanine-7-amido-4-trifluoro-methylcoumarine (l-/d-PheAFC) were synthesized and deployed for applications in aqueous systems. Several proteases and esterases were tested to detect suitable biocatalysts for the enzymatic hydrolysis of the coumarine substrates. With α-chymotrypsin and the esterase from porcine liver, every coumarine substrate was hydrolyzed in an aqueous system and the determined enzymatic parameters were compared. The enantioselectivity and the enantiomeric excess of the enzymatic reactions were investigated. In simultaneous applications of the l- and d-substrates, the reactions were monitored on-line and the enantioselectivities, enantiomeric excesses and kinetic parameters were examined. To display the versatility of the on-line method, the enzymatic reactions were transferred to the organic solvent toluene. After the optimisation of the biocatalysis in toluene, the kinetic parameters were measured and the ee-parameters for the substrates and products were examined in off-line and on-line investigations.
Keywords
- α-Chymotrypsin, 2D-fluorescence spectroscopy, Coumarines, Enantioselective hydrolysis, Esterase, On-line monitoring
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Chemical Engineering(all)
- Bioengineering
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
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In: Enzyme and microbial technology, Vol. 37, No. 7, 14.06.2005, p. 673-686.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Synthesis, test and application of chirale fluorescence substrates to evaluate enzymatic processes in different reaction media
AU - Knüttel, Torsten
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 - 2005/6/14
Y1 - 2005/6/14
N2 - A pseudo-enantiomeric reaction was developed which was monitored on-line via 2D-fluorescence spectroscopy. During enzymatic reactions the enantiomeric excess can be followed on-line. Fluorescence spectroscopic detectable substrates, l-/d-phenylalanine-7-amido-4-methylcoumarine (l-/d-PheAMC) and l-/d-phenylalanine-7-amido-4-trifluoro-methylcoumarine (l-/d-PheAFC) were synthesized and deployed for applications in aqueous systems. Several proteases and esterases were tested to detect suitable biocatalysts for the enzymatic hydrolysis of the coumarine substrates. With α-chymotrypsin and the esterase from porcine liver, every coumarine substrate was hydrolyzed in an aqueous system and the determined enzymatic parameters were compared. The enantioselectivity and the enantiomeric excess of the enzymatic reactions were investigated. In simultaneous applications of the l- and d-substrates, the reactions were monitored on-line and the enantioselectivities, enantiomeric excesses and kinetic parameters were examined. To display the versatility of the on-line method, the enzymatic reactions were transferred to the organic solvent toluene. After the optimisation of the biocatalysis in toluene, the kinetic parameters were measured and the ee-parameters for the substrates and products were examined in off-line and on-line investigations.
AB - A pseudo-enantiomeric reaction was developed which was monitored on-line via 2D-fluorescence spectroscopy. During enzymatic reactions the enantiomeric excess can be followed on-line. Fluorescence spectroscopic detectable substrates, l-/d-phenylalanine-7-amido-4-methylcoumarine (l-/d-PheAMC) and l-/d-phenylalanine-7-amido-4-trifluoro-methylcoumarine (l-/d-PheAFC) were synthesized and deployed for applications in aqueous systems. Several proteases and esterases were tested to detect suitable biocatalysts for the enzymatic hydrolysis of the coumarine substrates. With α-chymotrypsin and the esterase from porcine liver, every coumarine substrate was hydrolyzed in an aqueous system and the determined enzymatic parameters were compared. The enantioselectivity and the enantiomeric excess of the enzymatic reactions were investigated. In simultaneous applications of the l- and d-substrates, the reactions were monitored on-line and the enantioselectivities, enantiomeric excesses and kinetic parameters were examined. To display the versatility of the on-line method, the enzymatic reactions were transferred to the organic solvent toluene. After the optimisation of the biocatalysis in toluene, the kinetic parameters were measured and the ee-parameters for the substrates and products were examined in off-line and on-line investigations.
KW - α-Chymotrypsin
KW - 2D-fluorescence spectroscopy
KW - Coumarines
KW - Enantioselective hydrolysis
KW - Esterase
KW - On-line monitoring
UR - http://www.scopus.com/inward/record.url?scp=25444528238&partnerID=8YFLogxK
U2 - 10.1016/j.enzmictec.2004.04.026
DO - 10.1016/j.enzmictec.2004.04.026
M3 - Article
AN - SCOPUS:25444528238
VL - 37
SP - 673
EP - 686
JO - Enzyme and microbial technology
JF - Enzyme and microbial technology
SN - 0141-0229
IS - 7
ER -