Expression, purification, crystallization and preliminary X-ray diffraction analysis of Aspergillus terreus endo-β-1,4-glucanase from glycoside hydrolase family 12

Acta Crystallogr F Struct Biol Commun. 2014 Feb;70(Pt 2):267-70. doi: 10.1107/S2053230X13034936. Epub 2014 Jan 28.

Abstract

Endoglucanases are important enzymes that are involved in the modification and degradation of cellulose. Filamentous fungi such as Aspergillus terreus are effective biomass degraders in nature owing to their capacity to produce an enzymatic arsenal of glycoside hydrolases, including endoglucanase from glycoside hydrolase family 12 (GH12). The A. terreus GH12 endoglucanase was cloned and overexpressed in A. nidulans, purified and crystallized. A single crystal was obtained from a solution consisting of 2 M ammonium sulfate, 5%(v/v) 2-propanol. X-ray diffraction data were collected to a resolution of 1.85 Å using synchrotron radiation and a preliminary molecular-replacement solution was obtained in the trigonal space group P3(2)21. The unit-cell parameters were a = b = 103.24, c = 48.96 Å.

Keywords: Aspergillus terreus; endo-β-1,4-glucanase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aspergillus / enzymology*
  • Base Sequence
  • Crystallization
  • Crystallography, X-Ray / methods*
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Glycoside Hydrolases / chemistry*
  • Glycoside Hydrolases / genetics
  • Glycoside Hydrolases / isolation & purification
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Protein Conformation
  • Sequence Homology, Amino Acid

Substances

  • DNA Primers
  • Glycoside Hydrolases