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Subcloning and nucleotide sequence of the 3,4‐dihydroxyphenylacetate (homoprotocatechuate) 2,3‐dioxygenase gene from Escherichia coli C
Author(s) -
Roper David I.,
Cooper Ronald A.
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)81437-s
Subject(s) - subcloning , dioxygenase , escherichia coli , nucleic acid sequence , open reading frame , peptide sequence , gene , biochemistry , biology , amino acid , microbiology and biotechnology , chemistry
A cloned gene encoding the Escherichia coli C homoprotocatechuate (HPC) dioxygenase, an aromatic ring cleavage enzyme, was used to produce large amounts of the protein. Preparations of E. coli C HPC dioxygenase, whether expressed from the cloned gene or produced by the bacterium, lost activity very rapidly. The pure protein showed one type of subunit of M 1 33000. The first 21 N‐terminal amino acids were sequenced and the data used to confirm that the open reading frame of 831 bp, identified from the nucleotide sequence, encoded HPC dioxygenase. Comparison of the derived amino acid sequence with those of other extradiol and intradiol dioxygenases showed no obvious similarity to any of them.