Open AccessExpression, crystallization and preliminary X‐ray crystallographic analysis of cellobiose 2‐epimerase from Dictyoglomus turgidum DSM 6724
Author(s) -
Pham TanViet,
Hong SeungHye,
Hong Myoungki,
Ngo HoPhuongThuy,
Oh DeokKun,
Kang LinWoo
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113024391
Subject(s) - cellobiose , monoclinic crystal system , crystallography , crystallization , chemistry , substrate (aquarium) , solvent , resolution (logic) , x ray crystallography , enzyme , stereochemistry , crystal structure , diffraction , biochemistry , biology , organic chemistry , cellulase , physics , ecology , artificial intelligence , computer science , optics
Cellobiose 2‐epimerase epimerizes and isomerizes β‐1,4‐ and α‐1,4‐gluco‐oligosaccharides. N ‐Acyl‐D‐glucosamine 2‐epimerase (DT_epimerase) from Dictyoglomus turgidum has an unusually high catalytic activity towards its substrate cellobiose. DT_epimerase was expressed, purified and crystallized. Crystals were obtained of both His‐tagged DT_epimerase and untagged DT_epimerase. The crystals of His‐tagged DT_epimerase diffracted to 2.6 Å resolution and belonged to the monoclinic space group P 2 1 , with unit‐cell parameters a = 63.9, b = 85.1, c = 79.8 Å, β = 110.8°. With a Matthews coefficient V M of 2.18 Å 3 Da −1 , two protomers were expected to be present in the asymmetric unit with a solvent content of 43.74%. The crystals of untagged DT_epimerase diffracted to 1.85 Å resolution and belonged to the orthorhombic space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 55.9, b = 80.0, c = 93.7 Å. One protomer in the asymmetric unit was expected, with a corresponding V M of 2.26 Å 3 Da −1 and a solvent content of 45.6%.