Cloning, expression, purification and crystallization of saccharopine reductase from Magnaporthe grisea

Acta Crystallogr D Biol Crystallogr. 2000 May;56(Pt 5):662-4. doi: 10.1107/s0907444900003735.

Abstract

The gene coding for saccharopine reductase (E.C. 1.5.1.10), an enzyme of the alpha-aminoadipic pathway of lysine biosynthesis in the pathogenic fungus Magnaporthe grisea, was cloned and expressed in Escherichia coli. The purified enzyme was crystallized in space groups C2 and C222(1) using ammonium sulfate pH 4.8 or PEG 6000 pH 4. 1 as precipitants. The unit-cell parameters are a = 115.0, b = 56.6, c = 74.3 A, beta = 111.1 degrees for space group C2, and a = 89.3, b = 119.0, c = 195.9 A for space group C222(1). The crystals diffract to resolutions of 2.0 A (C2) and 2.4 A (C222(1)) at synchrotron sources.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Crystallization
  • Escherichia coli
  • Magnaporthe / enzymology*
  • Magnaporthe / genetics
  • Molecular Sequence Data
  • Open Reading Frames
  • Polymerase Chain Reaction
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Saccharopine Dehydrogenases / chemistry*
  • Saccharopine Dehydrogenases / genetics*
  • Saccharopine Dehydrogenases / isolation & purification

Substances

  • Recombinant Proteins
  • Saccharopine Dehydrogenases
  • saccharopine dehydrogenase (NADP, L-glutamate-forming)

Associated data

  • GENBANK/AF144424