Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the glutamate-1-semialdehyde aminotransferase from Bacillus subtilis

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 May 1;62(Pt 5):483-5. doi: 10.1107/S1744309106013121. Epub 2006 Apr 28.

Abstract

5-Aminolevulinic acid (ALA) is the first committed universal precursor in the tetrapyrrole-biosynthesis pathway. Plants, algae and many other bacteria synthesize ALA from glutamate by a C5 pathway in which the carbon skeleton of glutamate is converted into ALA by a series of enzymes. Glutamate-1-semialdehyde aminotransferase (GSAT) is the last enzyme in this pathway. The gene that codes for GSAT was amplified from the cDNA library of Bacillus subtilis and overexpressed in Escherichia coli strain BL21(DE3). The protein was purified and crystallized. Well diffracting single crystals were obtained by the hanging-drop vapour-diffusion method. Preliminary X-ray diffraction studies yielded excellent diffraction data to a resolution of 2.0 angstroms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis / enzymology*
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / metabolism
  • Intramolecular Transferases / chemistry*
  • Intramolecular Transferases / genetics*
  • Intramolecular Transferases / isolation & purification

Substances

  • Intramolecular Transferases
  • glutamate-1-semialdehyde 2,1-aminomutase