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

Min Kyu Kim, Young Jun An, Chang Sook Jeong, Jung Min Song, Mee Hye Kang, Youn Ho Lee, Sun Shin Cha

Research output: Contribution to journalArticle

2 Scopus citations

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 0.1M Tris-HCl pH 8.5, 25%(w/v) polyethylene glycol 3350 by the microbatch crystallization method at 295K. The CaMan protein crystal belonged to space group P212121, with unit-cell parameters a = 73.40, b = 83.81, c = 163.55Å. 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 1mM M5.

Original languageEnglish (US)
Pages (from-to)1007-1010
Number of pages4
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume69
Issue number9
DOIs
StatePublished - Sep 2013

Keywords

  • Cryptopygus antarcticus
  • glycoside hydrolase family 5
  • β-1,4-d-mannanase

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