Molecular characterisation of a thermoactive beta-1,3-glucanase from Oerskovia xanthineolytica

J Parrado, P R Escuredo, F Conejero-Lara, M Kotik, C P Ponting, J A Asenjo, C M Dobson

Research output: Contribution to journalArticlepeer-review

Abstract

Molecular characterisation of a lytic thermoactive beta-1,3-glucanase from Oerskovia xanthineolytica LL-G109 has been performed. A molecular mass of 27 195.6 +/- 1.3 Da and an isoelectric point of 4.85 were determined by electrospray mass spectrometry and from its titration curve, respectively. Its thermoactivity profile shows it to be a heat-stable enzyme with a temperature optimum of 65 degrees C. The secondary structure content of the protein was estimated by circular dichroism to be approx. 25% alpha-helix, 7% random coil, and 68% beta-sheet and beta-turn structure. Nuclear magnetic resonance spectra confirm the high content of beta-structure. Furthermore, the presence of a compact hydrophobic core is indicated by the presence of slowly exchanging amide hydrogens and the enzyme's relatively high resistance to proteolysis. The N-terminal sequences of the intact protein and of a tryptic peptide each exhibit significant similarity to family 16 of glycosyl hydrolases whose overall fold is known to contain almost exclusively beta-sheets and surface loops. Moreover, the sequenced tryptic peptide appears to encompass residues of the Oerskovia xanthineolytica glucanase active site, since it contains a portion of the family 16 active-site motif E-[L/I/V]-D-[L/I/V]-E.

Original languageEnglish
Pages (from-to)145-51
Number of pages7
JournalBBA - Bioenergetics
Volume1296
Issue number2
Publication statusPublished - 5 Sep 1996

Keywords

  • Amino Acid Sequence
  • Binding Sites
  • Circular Dichroism
  • Fungal Proteins
  • Glucan 1,3-beta-Glucosidase
  • Hot Temperature
  • Isoelectric Point
  • Magnetic Resonance Spectroscopy
  • Mass Spectrometry
  • Molecular Sequence Data
  • Molecular Weight
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Yeasts
  • beta-Glucosidase

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