Expression, purification and preliminary crystallographic studies of alpha-amylase isozyme 1 from barley seeds

Acta Crystallogr D Biol Crystallogr. 2002 Apr;58(Pt 4):683-6. doi: 10.1107/s090744490200166x. Epub 2002 Mar 22.

Abstract

The germinating barley seed contains two major alpha-amylase isozyme families, AMY1 and AMY2, involved in starch degradation to provide energy used by the plant embryo for growth. Many years of difficulty in growing three-dimensional crystals of natural AMY1 have now been overcome by a nonapeptide truncation of the enzyme C-terminus. The truncated enzyme was overexpressed in Pichia pastoris, purified and crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol 8000 as precipitant and 2-propanol as an additive. Crystals belong to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 88.36, b = 72.82, c = 61.74 A and one molecule per asymmetric unit.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallography, X-Ray
  • Hordeum / chemistry*
  • Isoenzymes / chemistry
  • Pichia / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Seeds / chemistry
  • alpha-Amylases / chemistry*
  • alpha-Amylases / isolation & purification
  • alpha-Amylases / metabolism

Substances

  • Isoenzymes
  • Recombinant Proteins
  • alpha-Amylases