Use of gene fusions of the structural gene sdaA to purify L-serine deaminase 1 from Escherichia coli K-12

Eur J Biochem. 1993 Feb 1;211(3):521-7. doi: 10.1111/j.1432-1033.1993.tb17578.x.

Abstract

The purification by affinity chromatography of beta-galactosidase from strains carrying sdaA/lacZ gene fusions results in the copurification of L-serine deaminase 1. We conclude that sdaA is the structural gene for the latter enzyme. The purified L-serine deaminase 1 obtained after collagenase treatment of an sdaA-collagen-lacZ fusion differs from the native enzyme by the addition of several amino acids at the C-terminal. Like the enzyme in crude extracts, this purified enzyme is catalytically inactive, and is activated by incubation with iron and dithiothreitol.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular*
  • Collagen / genetics
  • Collagenases / metabolism
  • Dithiothreitol / pharmacology
  • Enzyme Activation / drug effects
  • Escherichia coli / enzymology
  • Escherichia coli / genetics*
  • Genes, Bacterial*
  • Iron / pharmacology
  • L-Serine Dehydratase / genetics
  • L-Serine Dehydratase / isolation & purification*
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Plasmids
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • beta-Galactosidase / genetics
  • beta-Galactosidase / isolation & purification

Substances

  • Recombinant Fusion Proteins
  • Collagen
  • Iron
  • beta-Galactosidase
  • Collagenases
  • L-Serine Dehydratase
  • Dithiothreitol

Associated data

  • GENBANK/L09679
  • GENBANK/L09680
  • GENBANK/L09681
  • GENBANK/L09682
  • GENBANK/L09683
  • GENBANK/L16554
  • GENBANK/L16555
  • GENBANK/L16556
  • GENBANK/M28695
  • GENBANK/Z18286