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

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

Journal Name: Protein & Peptide Letters

Volume 15 , Issue 10 , 2008

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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.

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

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Article Details

VOLUME: 15
ISSUE: 10
Year: 2008
Page: [1138 - 1141]
Pages: 4
DOI: 10.2174/092986608786071201
Price: $58

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