Use of gene fusions of the structural gene sdaA to purify L-serine deaminase 1 from Escherichia coli K-12.
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.
Su
H
H
Biology Department, Concordia University, Montreal, Canada.
Moniakis
J
J
Newman
E B
EB
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics; D017364:Collagenases / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial; D007501:Iron / Q000494:pharmacology; D012695:L-Serine Dehydratase / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000302:isolation & purification
D011993:Recombinant Fusion Proteins; D003094:Collagen; D007501:Iron; D001616:beta-Galactosidase; D017364:Collagenases; D012695:L-Serine Dehydratase; D004229:Dithiothreitol