Open AccessCloning, recombinant production, crystallization and preliminary X‐ray diffraction studies of a family 84 glycoside hydrolase from Clostridium perfringens
Author(s) -
FickoBlean Elizabeth,
Boraston Alisdair B.
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105024012
Subject(s) - clostridium perfringens , escherichia coli , recombinant dna , crystallization , cloning (programming) , clostridium , biology , microbiology and biotechnology , biochemistry , chemistry , bacteria , gene , genetics , organic chemistry , computer science , programming language
Clostridium perfringens is a ubiquitous environmental organism that is capable of causing a variety of diseases in mammals, including gas gangrene and necrotic enteritis in humans. The activity of a secreted hyaluronidase, attributed to the NagH protein, contributes to the pathogenicity of this organism. The family 84 catalytic module of one of the three homologues of NagH found in C. perfringens (ATCC 13124) has been cloned. The 69 kDa catalytic module of NagJ, here called GH84C, was overproduced in Escherichia coli and purified by immobilized metal‐affinity chromatography (IMAC). Crystals belonging to space group I 222 or I 2 1 2 1 2 1 with unit‐cell parameters a = 130.39, b = 150.05, c = 155.43 Å were obtained that diffracted to 2.1 Å. Selenomethionyl crystals have also been produced, leading to the possibility of solving the phase problem by MAD using synchrotron radiation.