title: 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.
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.
authors.affiliation: Marine Biotechnology Research Division, Korea Institute of Ocean Science and Technology, Ansan 426-744, Republic of Korea.
authors.forename: Min-Kyu
authors.initials: MK
authors.lastname: Kim
authors.forename: Young Jun
authors.initials: YJ
authors.lastname: An
authors.forename: Chang-Sook
authors.initials: CS
authors.lastname: Jeong
authors.forename: Jung Min
authors.initials: JM
authors.lastname: Song
authors.forename: Mee Hye
authors.initials: MH
authors.lastname: Kang
authors.forename: Youn-Ho
authors.initials: YH
authors.lastname: Lee
authors.forename: Sun-Shin
authors.initials: SS
authors.lastname: Cha
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000864:Antarctic Regions; D060829:Arthropod Proteins / Q000737:chemistry* / Q000235:genetics; D001181:Arthropods / Q000737:chemistry* / Q000201:enzymology / Q000235:genetics; D003080:Cold Temperature; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008361:Mannosidases / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment
chemical_list: D060829:Arthropod Proteins; D011994:Recombinant Proteins; D008361:Mannosidases; C412205:beta-1,4-mannosidase
keywords: Cryptopygus antarcticus; glycoside hydrolase family 5; β-1,4-d-mannanase