Open AccessCloning, expression, purification, crystallization and preliminary crystallographic studies of UgdG, an UDP‐glucose dehydrogenase from Sphingomonas elodea ATCC 31461
Author(s) -
Rocha Joana,
Granja Ana Teresa,
SáCorreia Isabel,
Fialho Arsénio,
Frazão Carlos
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910904929x
Subject(s) - cloning (programming) , crystallization , chemistry , biochemistry , crystallography , biology , microbiology and biotechnology , organic chemistry , computer science , programming language
Gellan gum, a commercial gelling agent produced by Sphingomonas elodea ATCC 31461, is a high‐value microbial exopolysaccharide. UDP‐glucose dehydrogenase (UGD; EC 1.1.1.22) is responsible for the NAD‐dependent twofold oxidation of UDP‐glucose to UDP‐glucuronic acid, one of the key components for gellan biosynthesis. S. elodea ATCC 31461 UGD, termed UgdG, was cloned, expressed, purified and crystallized in native and SeMet‐derivatized forms in hexagonal and tetragonal space groups, respectively; the crystals diffracted X‐rays to 2.40 and 3.40 Å resolution, respectively. Experimental phases were obtained for the tetragonal SeMet‐derivatized crystal form by a single‐wavelength anomalous dispersion experiment. This structure was successfully used as a molecular‐replacement probe for the hexagonal crystal form of the native protein.