Purification and analysis of the structure of α-galactosidase from Escherichiacoli

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Abstract

α-Galactosidase, the product of the melA gene, was purified from a strain of Escherichiacoli harboring a plasmid carrying melA, which overproduced the α-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.

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