Chiral Hydroxylation at the Mononuclear Nonheme Fe(II) Center of 4-(S) Hydroxymandelate Synthase - A Structure-Activity Relationship Analysis

Cristiana M.L. Di Giuro, Cornelia Konstantinovics, Uwe Rinner, Christina Nowikow, Erich Leitner, Grit D. Straganz

Research output: Contribution to journalArticle

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Abstract

(S)-Hydroxymandelate synthase (Hms) is a nonheme Fe(II) dependent dioxygenase that catalyzes the oxidation of 4-hydroxyphenylpyruvate to (S)-4-hydroxymandelate by molecular oxygen. In this work, the substrate promiscuity of Hms is characterized in order to assess its potential for the biosynthesis of chiral α-hydroxy acids. Enzyme kinetic analyses, the characterization of product spectra, quantitative structure activity relationship (QSAR) analyses and in silico docking studies are used to characterize the impact of substrate properties on particular steps of catalysis. Hms is found to accept a range of α-oxo acids, whereby the presence of an aromatic substituent is crucial for efficient substrate turnover. A hydrophobic substrate binding pocket is identified as the likely determinant of substrate specificity. Upon introduction of a steric barrier, which is suspected to obstruct the accommodation of the aromatic ring in the hydrophobic pocket during the final hydroxylation step, the racemization of product is obtained. A steady state kinetic analysis reveals that the turnover number of Hms strongly correlates with substrate hydrophobicity. The analysis of product spectra demonstrates high regioselectivity of oxygenation and a strong coupling efficiency of C-C bond cleavage and subsequent hydroxylation for the tested substrates. Based on these findings the structural basis of enantioselectivity and enzymatic activity is discussed.

Original languageEnglish
Article numbere68932
JournalPLoS One
Volume8
Issue number7
DOIs
Publication statusPublished - Jul 23 2013

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Hydroxylation
hydroxylation
structure-activity relationships
Structure-Activity Relationship
aromatic compounds
quantitative structure-activity relationships
Keto Acids
Dioxygenases
enzyme kinetics
Hydroxy Acids
Quantitative Structure-Activity Relationship
acids
enantiomers
Substrates
substrate specificity
Substrate Specificity
hydrophobicity
Catalysis
Hydrophobic and Hydrophilic Interactions
catalytic activity

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Chiral Hydroxylation at the Mononuclear Nonheme Fe(II) Center of 4-(S) Hydroxymandelate Synthase - A Structure-Activity Relationship Analysis. / Di Giuro, Cristiana M.L.; Konstantinovics, Cornelia; Rinner, Uwe; Nowikow, Christina; Leitner, Erich; Straganz, Grit D.

In: PLoS One, Vol. 8, No. 7, e68932, 23.07.2013.

Research output: Contribution to journalArticle

Di Giuro, Cristiana M.L. ; Konstantinovics, Cornelia ; Rinner, Uwe ; Nowikow, Christina ; Leitner, Erich ; Straganz, Grit D. / Chiral Hydroxylation at the Mononuclear Nonheme Fe(II) Center of 4-(S) Hydroxymandelate Synthase - A Structure-Activity Relationship Analysis. In: PLoS One. 2013 ; Vol. 8, No. 7.
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