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

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Mar 1;68(Pt 3):328-9. doi: 10.1107/S1744309112003740. Epub 2012 Feb 23.

Abstract

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).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cellulase / chemistry*
  • Crystallization
  • Crystallography, X-Ray
  • Enzyme Stability
  • Pyrococcus furiosus / enzymology*

Substances

  • Cellulase