Crystallization and preliminary X-ray analysis of a hyperthermophilic endoglucanase from Pyrococcus furiosus.

The hyperthermophilic glycoside hydrolase family 12 endocellulase from the archaeon Pyrococcus furiosus (EGPf) catalyzes the hydrolytic cleavage of the β-1,4-glucosidic linkage in β-glucans in biomass. EGPf (Gene ID PF0854; EC 3.2.1.4) contains a signal sequence and proline- and hydroxyl-rich regions at the N-terminus. Truncated EGPf (EGPfΔN30) without the proline- and hydroxyl-rich regions at the N-terminus was prepared and subjected to crystallization experiments. Crystals were obtained using the hanging-drop vapour-diffusion method at 303 K. An X-ray diffraction data set was collected to 1.07 Å resolution at 100 K. The crystal belonged to space group P2(1)2(1)2, with unit-cell parameters a = 58.01, b = 118.67, c = 46.76 Å. The presence of one molecule of enzyme per asymmetric unit gives a crystal volume per protein mass (V(M)) of 2.63 Å(3) Da(-1) and a solvent content of 53.3%(v/v).