Open AccessExpression, crystallization and preliminary X‐ray diffraction studies of recombinant Clostridium perfringens β2‐toxin
Author(s) -
Gurjar Abhijit A.,
Yennawar Neela H.,
Yennawar Hemant P.,
Rajashankar Kanagalaghatta R.,
Hegde Narasimha V.,
Jayarao Bhushan M.
Publication year - 2007
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309107020313
Subject(s) - clostridium perfringens , toxin , crystallization , recombinant dna , exotoxin , microbiology and biotechnology , clostridium , escherichia coli , biology , bacteria , chemistry , crystallography , gene , biochemistry , genetics , organic chemistry
Clostridium perfringens is a Gram‐positive sporulating anaerobic bacterium that is responsible for a wide spectrum of diseases in animals, birds and humans. The virulence of C. perfringens is associated with the production of several enterotoxins and exotoxins. β2‐toxin is a 28 kDa exotoxin produced by C. perfringens . It is implicated in necrotic enteritis in animals and humans, a disease characterized by a sudden acute onset with lethal hemorrhagic mucosal ulceration. The recombinant expression, purification and crystallization of β2‐toxin using the batch‐under‐oil technique are reported here. Native X‐ray diffraction data were obtained to 2.9 Å resolution on a synchrotron beamline at the F2 station at Cornell High Energy Synchrotron Source (CHESS) using an ADSC Quantum‐210 CCD detector. The crystals belong to space group R 3, with a dimer in the asymmetric unit; the unit‐cell parameters are a = b = 103.71, c = 193.48 Å, α = β = 90, γ = 120° using the hexagonal axis setting. A self‐rotation function shows that the two molecules are related by a noncrystallographic twofold axis with polar angles ω = 90.0, ϕ = 210.3°.