4URF

Molecular Genetic and Crystal Structural Analysis of 1-(4- Hydroxyphenyl)-Ethanol Dehydrogenase from Aromatoleum aromaticum EbN1

Summary for 4URF

• Entry DOI: 10.2210/pdb4urf/pdb
• Related: 4URE
• Descriptor: CYCLOHEXANOL DEHYDROGENASE, NICOTINAMIDE-ADENINE-DINUCLEOTIDE, ACETATE ION, ... (7 entities in total)
• Functional Keywords: oxidoreductase, short chain dehydrogenase, aromatic compounds, anaerobic degradation, alcohol dehydrogenase, molecular genetics, stereochemistry
• Biological source: AROMATOLEUM AROMATICUM EBN1
• Total number of polymer chains: 2
• Total formula weight: 54808.29

Authors

Buesing, I.,Hoeffken, H.W.,Breuer, M.,Woehlbrand, L.,Hauer, B.,Rabus, R. (deposition date: 2014-06-28, release date: 2015-07-08, Last modification date: 2024-01-10)

Primary citation

Busing, I.,Hoffken, H.W.,Breuer, M.,Wohlbrand, L.,Hauer, B.,Rabus, R.
Molecular Genetic and Crystal Structural Analysis of 1-(4-Hydroxyphenyl)-Ethanol Dehydrogenase from 'Aromatoleum Aromaticum' Ebn1.
J.Mol.Microbiol.Biotechnol., 25:327-, 2015

PubMed Abstract: The dehydrogenation of 1-(4-hydroxyphenyl)-ethanol to 4-hydroxyacetophenone represents the second reaction step during anaerobic degradation of p-ethylphenol in the denitrifying bacterium 'Aromatoleum aromaticum' EbN1. Previous proteogenomic studies identified two different proteins (ChnA and EbA309) as possible candidates for catalyzing this reaction [Wöhlbrand et al: J Bacteriol 2008;190:5699-5709]. Physiological-molecular characterization of newly generated unmarked in-frame deletion and complementation mutants allowed defining ChnA (renamed here as Hped) as the enzyme responsible for 1-(4-hydroxyphenyl)-ethanol oxidation. Hped [1-(4-hydroxyphenyl)-ethanol dehydrogenase] belongs to the 'classical' family within the short-chain alcohol dehydrogenase/reductase (SDR) superfamily. Hped was overproduced in Escherichia coli, purified and crystallized. The X-ray structures of the apo- and NAD(+)-soaked form were resolved at 1.5 and 1.1 Å, respectively, and revealed Hped as a typical homotetrameric SDR. Modeling of the substrate 4-hydroxyacetophenone (reductive direction of Hped) into the active site revealed the structural determinants of the strict (R)-specificity of Hped (Phe(187)), contrasting the (S)-specificity of previously reported 1-phenylethanol dehydrogenase (Ped; Tyr(93)) from strain EbN1 [Höffken et al: Biochemistry 2006;45:82-93].

PubMed: 26488297
DOI: 10.1159/000439113

Experimental method

X-RAY DIFFRACTION (1.1 Å)