Crystallization and X‐ray diffraction analysis of a putative bacterial class I labdane‐related diterpene synthase. Issue 9 (1st September 2015)

• Record Type: Journal Article
• Authors: Serrano‐Posada, Hugo; Centeno‐Leija, Sara; Rojas‐Trejo, Sonia; Stojanoff, Vivian; Rodríguez‐Sanoja, Romina; Rudiño‐Piñera, Enrique; Sánchez, Sergio
• Is Part Of: Acta crystallographica. Volume 71:Issue 9(2015:Sep.)
• Page Start: 1194
• Page End: 1199
• Publication Date: 2015-09-01
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X15014363
• Languages: English
• ISSNs: 2053-230X

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Labdane‐related diterpenoids are natural products with potential pharmaceutical applications that are rarely found in bacteria. Here, a putative class I labdane‐related diterpene synthase (LrdC) identified by genome mining in a streptomycete was successfully crystallized using the microbatch method. Crystals of the LrdC enzyme were obtained in a holo form with its natural cofactor Mg<sup>2+</sup> (LrdC‐Mg<sup>2+</sup>) and in complex with inorganic pyrophosphate (PP<sub>i</sub>) (LrdC‐Mg<sup>2+</sup>–PP<sub>i</sub>). Crystals of native LrdC‐Mg<sup>2+</sup> diffracted to 2.50 Å resolution and belonged to the trigonal space group <italic>P</italic>3<sub>2</sub>21, with unit‐cell parameters <italic>a</italic> = <italic>b</italic> = 107.1, <italic>c</italic> = 89.2 Å. Crystals of the LrdC‐Mg<sup>2+</sup>–PP<sub>i</sub> complex grown in the same conditions as the native enzyme with PEG 8000 diffracted to 2.36 Å resolution and also belonged to the trigonal space group <italic>P</italic>3<sub>2</sub>21. Crystals of the LrdC‐Mg<sup>2+</sup>–PP<sub>i</sub> complex grown in a second crystallization condition with PEG 3350 diffracted to 2.57 Å resolution and belonged to the monoclinic space group <italic>P</italic>2<sub>1</sub>, with unit‐cell parameters <italic>a</italic> = 49.9, <italic>b</italic> = 104.1, <italic>c</italic> = 66.5 Å, β = 111.4°. The structure was determined by the single‐wavelength anomalous dispersion (SAD) technique using the osmium signal from a potassium hexachloroosmate (IV) derivative.</p> </abstract>