Crystallization of recombinant chitinase from the cloned chiA gene of Serratia marcescens.
The chiA gene encoding for the chitinase enzyme from Serratia marcescens was efficiently overexpressed under the pL promoter and the enzyme was secreted into the growth medium. The chitinase was purified to homogeneity using affinity chromatography on a Phenyl-Sepharose column and the protein was successfully crystallized. The crystals are presently in the form of small needles in space group C222(1) and have unit cell dimensions a = 204(+/- 0.5) A, b = 134(+/- 0.5) A, c = 60(+/- 0.5) A. The crystals diffract X-rays to about 3 A resolution and are suitable for three-dimensional structural analysis.
Vorgias
C E
CE
European Molecular Biology Laboratory, DESY, Hamburg, Germany.
Kingswell
A J
AJ
Dauter
Z
Z
Oppenheim
A B
AB
D002688:Chitinases / Q000737:chemistry / Q000302:isolation & purification*; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012706:Serratia marcescens / Q000201:enzymology* / Q000235:genetics; D014961:X-Ray Diffraction
D011994:Recombinant Proteins; D002688:Chitinases