Cloning, Expression, Purification, Crystallization and Preliminary X-Ray Crystallographic Analysis of C-12 Hydroxylase EryK from Saccharopolyspora erythraea

Title: Cloning, Expression, Purification, Crystallization and Preliminary X-Ray Crystallographic Analysis of C-12 Hydroxylase EryK from Saccharopolyspora erythraea

Volume: 15 Issue: 10

Author(s): Carmelinda Savino, Giuliano Sciara, Adriana E. Miele, Steven G. Kendrew and Beatrice Vallone

Affiliation:

Keywords: Crystallization, erythromycin, cytochrome P450, C-12 hydroxylase, x-ray

Abstract: Erythromycin A is produced by Saccharopolyspora erythraea via a secondary metabolic pathway using several steps including glycosylations and hydroxylations of the first macrolide intermediate 6-deoxyerythronolide B. Erythromycin C-12 hydroxylase (CYP113A1), the P450 cytochrome active in the final stages of erythromycin biosynthesis, was cloned and expressed in E. coli. Different crystal forms were harvested from distinct crystallization conditions: two ligandfree forms, one substrate bound and two inhibitors-bound. All crystals belong either to the monoclinc P21 or to the orthorhombic P212121 space groups, and exhibit diffraction limits ranging from 2.3 to 1.6 Å. The structures will be determined by molecular replacement.

Cite this article as:

Savino Carmelinda, Sciara Giuliano, Miele E. Adriana, Kendrew G. Steven and Vallone Beatrice, Cloning, Expression, Purification, Crystallization and Preliminary X-Ray Crystallographic Analysis of C-12 Hydroxylase EryK from Saccharopolyspora erythraea, Protein & Peptide Letters 2008; 15 (10) . https://dx.doi.org/10.2174/092986608786071201