Purification, crystallization and preliminary X-ray crystallographic analysis of 3-ketosteroid Δ 1-dehydrogenase from Rhodococcus erythropolis SQ1

Ali Rohman, Niels Van Oosterwijk, Bauke W. Dijkstra

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

3-Ketosteroid Δ 1-dehydrogenase plays a crucial role in the early steps of steroid degradation by introducing a double bond between the C1 and C2 atoms of the A-ring of its 3-ketosteroid substrates. The 3-ketosteroid Δ 1-dehydrogenase from Rhodococcus erythropolis SQ1, a 56 kDa flavoprotein, was crystallized using the sitting-drop vapour-diffusion method at room temperature. The crystals grew in various buffers over a wide pH range (from pH 5.5 to 10.5), but the best crystallization condition consisted of 2%(v/v) PEG 400, 0.1 M HEPES pH 7.5, 2.0 M ammonium sulfate. A native crystal diffracted X-rays to 2.0 Å resolution. It belonged to the primitive orthorhombic space group P212121, with unit-cell parameters a = 107.4, b = 131.6, c = 363.2 Å, and contained eight molecules in the asymmetric unit. The initial structure of the enzyme was solved using multi-wavelength anomalous dispersion (MAD) data collected from a Pt-derivatized crystal.

Original languageEnglish
Pages (from-to)551-556
Number of pages6
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume68
Issue number5
DOIs
Publication statusPublished - May 2012

Keywords

  • 3-ketosteroid Δ -dehydrogenase
  • Rhodococcus erythropolis SQ1

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