Cloning, expression, purification, crystallization and preliminary crystallographic studies of UgdG, an UDP-glucose dehydrogenase from Sphingomonas elodea ATCC 31461.
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 A 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.
Rocha
Joana
J
Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Apartado 127, 2781-901 Oeiras, Portugal.
Granja
Ana Teresa
AT
Sá-Correia
Isabel
I
Fialho
Arsénio
A
Frazão
Carlos
C
D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D012645:Selenomethionine / Q000378:metabolism; D020578:Sphingomonas; D014533:Uridine Diphosphate Glucose Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification
D012645:Selenomethionine; D014533:Uridine Diphosphate Glucose Dehydrogenase