Details
| Original language | English |
|---|---|
| Pages (from-to) | 18-26 |
| Number of pages | 9 |
| Journal | Biotechnology and bioengineering |
| Volume | 106 |
| Issue number | 1 |
| Early online date | 31 Dec 2009 |
| Publication status | Published - 1 May 2010 |
| Externally published | Yes |
Abstract
Benzoylformate decarboxylase (BFD) from Pseudomonas putida is a thiamine diphosphate-dependent (ThDP) enzyme that catalyzes the asymmetric C-C bond formation to (S)-2-hydroxypropiophenone [(S)-HPP] starting from benzaldehyde and acetaldehyde. The enantioselectivity of BFD was shown to be a function of temperature and substrate concentration. It can additionally be changed by site-directed mutagenesis on hot spot positions in the active site. In this article, we present the effect of hydrostatic pressure up to 250 MPa on the enantioselectivity for the recombinant wtBFD as well as for the variants BFD F464I, BFD A460I, and BFD A460I-F464I. A general tendency toward lower amounts of (S)-HPP could be observed at increasing pressures. For two of these variants an increase in pressure even caused an inversion in the enantioselectivity and thus increasing enantiomeric excesses, respectively. A pressure-induced increase in enantioselectivity could therefore be observed for the first time in biocatalysis to the best of our knowledge. Furthermore, the pH is shown to be a parameter that also significantly influences the enantioselectivity of the reaction mentioned above.
Keywords
- Benzoylformate decarboxylase, C-C bond formation, Enantioselectivity, Enzyme catalysis, High pressure, pH
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Chemical Engineering(all)
- Bioengineering
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
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In: Biotechnology and bioengineering, Vol. 106, No. 1, 01.05.2010, p. 18-26.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Influence of the hydrostatic pressure and pH on the asymmetric 2-hydroxyketone formation catalyzed by pseudomonas putida benzoylformate decarboxylase and variants thereof
AU - Berheide, Marco
AU - Peper, Stephanie
AU - Kara, Selin
AU - Long, Wei Sing
AU - Schenkel, Steffen
AU - Pohl, Martina
AU - Niemeyer, Bernd
AU - Liese, Andreas
PY - 2010/5/1
Y1 - 2010/5/1
N2 - Benzoylformate decarboxylase (BFD) from Pseudomonas putida is a thiamine diphosphate-dependent (ThDP) enzyme that catalyzes the asymmetric C-C bond formation to (S)-2-hydroxypropiophenone [(S)-HPP] starting from benzaldehyde and acetaldehyde. The enantioselectivity of BFD was shown to be a function of temperature and substrate concentration. It can additionally be changed by site-directed mutagenesis on hot spot positions in the active site. In this article, we present the effect of hydrostatic pressure up to 250 MPa on the enantioselectivity for the recombinant wtBFD as well as for the variants BFD F464I, BFD A460I, and BFD A460I-F464I. A general tendency toward lower amounts of (S)-HPP could be observed at increasing pressures. For two of these variants an increase in pressure even caused an inversion in the enantioselectivity and thus increasing enantiomeric excesses, respectively. A pressure-induced increase in enantioselectivity could therefore be observed for the first time in biocatalysis to the best of our knowledge. Furthermore, the pH is shown to be a parameter that also significantly influences the enantioselectivity of the reaction mentioned above.
AB - Benzoylformate decarboxylase (BFD) from Pseudomonas putida is a thiamine diphosphate-dependent (ThDP) enzyme that catalyzes the asymmetric C-C bond formation to (S)-2-hydroxypropiophenone [(S)-HPP] starting from benzaldehyde and acetaldehyde. The enantioselectivity of BFD was shown to be a function of temperature and substrate concentration. It can additionally be changed by site-directed mutagenesis on hot spot positions in the active site. In this article, we present the effect of hydrostatic pressure up to 250 MPa on the enantioselectivity for the recombinant wtBFD as well as for the variants BFD F464I, BFD A460I, and BFD A460I-F464I. A general tendency toward lower amounts of (S)-HPP could be observed at increasing pressures. For two of these variants an increase in pressure even caused an inversion in the enantioselectivity and thus increasing enantiomeric excesses, respectively. A pressure-induced increase in enantioselectivity could therefore be observed for the first time in biocatalysis to the best of our knowledge. Furthermore, the pH is shown to be a parameter that also significantly influences the enantioselectivity of the reaction mentioned above.
KW - Benzoylformate decarboxylase
KW - C-C bond formation
KW - Enantioselectivity
KW - Enzyme catalysis
KW - High pressure
KW - pH
UR - http://www.scopus.com/inward/record.url?scp=77951549438&partnerID=8YFLogxK
U2 - 10.1002/bit.22650
DO - 10.1002/bit.22650
M3 - Article
C2 - 20047192
AN - SCOPUS:77951549438
VL - 106
SP - 18
EP - 26
JO - Biotechnology and bioengineering
JF - Biotechnology and bioengineering
SN - 0006-3592
IS - 1
ER -