Document Detail

Influence of the hydrostatic pressure and pH on the asymmetric 2-hydroxyketone formation catalyzed by Pseudomonas putida benzoylformate decarboxylase and variants thereof.
MedLine Citation:
PMID:  20047192     Owner:  NLM     Status:  MEDLINE    
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.
Marco Berheide; Stephanie Peper; Selin Kara; Wei Sing Long; Steffen Schenkel; Martina Pohl; Bernd Niemeyer; Andreas Liese
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  106     ISSN:  1097-0290     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2010 May 
Date Detail:
Created Date:  2010-03-29     Completed Date:  2010-06-07     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  18-26     Citation Subset:  IM    
Institute of Technical Biocatalysis, Hamburg University of Technology, Denickestrasse 15, 21073 Hamburg, Germany.
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MeSH Terms
Acetaldehyde / metabolism
Amino Acid Substitution / genetics
Bacterial Proteins / metabolism*
Benzaldehydes / metabolism
Carboxy-Lyases / metabolism*
Hydrogen-Ion Concentration
Hydrostatic Pressure*
Hydroxypropiophenone / metabolism
Ketones / metabolism*
Mutant Proteins / metabolism
Pseudomonas putida / enzymology*
Recombinant Proteins / metabolism
Substrate Specificity
Reg. No./Substance:
0/Bacterial Proteins; 0/Benzaldehydes; 0/Ketones; 0/Mutant Proteins; 0/Recombinant Proteins; 100-52-7/benzaldehyde; 1321-48-8/Hydroxypropiophenone; 75-07-0/Acetaldehyde; EC 4.1.1.-/Carboxy-Lyases; EC decarboxylase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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