Purification and analysis of the structure of alpha-galactosidase from Escherichia coli.
Alpha-Galactosidase, the product of the melA gene, was purified from a strain of Escherichia coli harboring a plasmid carrying melA, which over-produced the alpha-galactosidase. An apparent molecular weight was determined to be 50 kDa. The amino acid composition of this enzyme was determined. The result indicates that this enzyme is a hydrophilic and acidic protein. We have subjected the purified enzyme to 20 cycles of N-terminal sequence analysis. This verified the translation start site of the melA gene and the predicted N-terminal sequence.
Nagao
Y
Y
Department of Microbiology, Faculty of Pharmaceutical Sciences, Okayama University, Japan.
Nakada
T
T
Imoto
M
M
Shimamoto
T
T
Sakai
S
S
Tsuda
M
M
Tsuchiya
T
T
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005696:Galactosidases / Q000302:isolation & purification*; D005798:Genes, Bacterial; D008970:Molecular Weight; D010957:Plasmids; D000519:alpha-Galactosidase / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*
D000596:Amino Acids; D005696:Galactosidases; D000519:alpha-Galactosidase