Expression at 279 K, purification, crystallization and preliminary X-ray crystallographic analysis of a novel cold-active β-1,4-D-mannanase from the Antarctic springtail Cryptopygus antarcticus

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Sep;69(Pt 9):1007-10. doi: 10.1107/S1744309113020538. Epub 2013 Aug 19.

Abstract

The CaMan gene product from Cryptopygus antarcticus, which belongs to the glycoside hydrolase family 5 type β-1,4-D-mannanases, has been crystallized using a precipitant solution consisting of 0.1 M Tris-HCl pH 8.5, 25%(w/v) polyethylene glycol 3350 by the microbatch crystallization method at 295 K. The CaMan protein crystal belonged to space group P212121, with unit-cell parameters a = 73.40, b = 83.81, c = 163.55 Å. Assuming the presence of two molecules in the asymmetric unit, the solvent content was estimated to be about 61.29%. CaMan-mannopentaose (M5) complex crystals that were isomorphous to the CaMan crystals were obtained using the same mother liquor containing 1 mM M5.

Keywords: Cryptopygus antarcticus; glycoside hydrolase family 5; β-1,4-d-mannanase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antarctic Regions
  • Arthropod Proteins / chemistry*
  • Arthropod Proteins / genetics
  • Arthropods / chemistry*
  • Arthropods / enzymology
  • Arthropods / genetics
  • Cold Temperature
  • Crystallography, X-Ray
  • Escherichia coli / genetics
  • Gene Expression
  • Mannosidases / chemistry*
  • Mannosidases / genetics
  • Molecular Sequence Data
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Sequence Alignment

Substances

  • Arthropod Proteins
  • Recombinant Proteins
  • Mannosidases
  • beta-1,4-mannosidase