Purification, crystallization and preliminary X-ray analysis of β-glucosidase from Kluyveromyces marxianus NBRC1777

Erina Yoshida, Masafumi Hidaka, Shinya Fushinobu, Takashi Koyanagi, Hiromichi Minami, Hisanori Tamaki, Motomitsu Kitaoka, Takane Katayama, Hidehiko Kumagai

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

The intracellular β-glucosidase from Kluyveromyces marxianus NBRC1777 (KmBglI) belongs to glycoside hydrolase family 3 and has a unique domain architecture. Selenomethionine-labelled KmBglI was purified and crystallized by the hanging-drop vapour-diffusion method using the purified enzyme at 30 mg ml-1, 0.04 M potassium dihydrogen phosphate pH 5.1, 16%(w/v) PEG 8000 and 20%(v/v) glycerol. The crystal belonged to space group C2, with unit-cell parameters a = 245.8, b = 148.7, c = 119.9 Å, β = 112.9°. Multiple-wavelength anomalous dispersion data were collected at 2.4 and 2.5 Å resolution. A tetramer was assumed to be present in the asymmetric unit, which gave a Matthews coefficient of 2.6 Å3 Da-1.

Original languageEnglish
Pages (from-to)1190-1192
Number of pages3
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume65
Issue number11
DOIs
Publication statusPublished - 2009
Externally publishedYes

Keywords

  • Glycoside hydrolase family 3
  • Kluyveromyces marxianus NBRC1777
  • β-glucosidases

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

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