Parathion hydrolase specified by the Flavobacterium opd gene: relationship between the gene and protein.
The sequence of a 1,693-base-pair plasmid DNA fragment from Flavobacterium sp. strain ATCC 27551 containing the parathion hydrolase gene (opd) was determined. Within this sequence, there is only one open reading frame large enough to encode the 35,000-dalton membrane-associated hydrolase protein purified from Flavobacterium extracts. Amino-terminal sequence analysis of the purified Flavobacterium hydrolase demonstrated that serine is the amino-terminal residue of the hydrolase protein. The amino-terminal serine corresponds to a TCG codon located 87 base pairs downstream of the presumptive ATG initiation codon in the nucleotide sequence. The amino acid composition of the purified protein agrees well with that predicted from the nucleotide sequence, using serine as the amino-terminal residue. These data suggest that the parathion hydrolase protein is processed at its amino terminus in Flavobacterium sp. Construction in Escherichia coli of a lacZ-opd gene fusion in which the first 33 amino-terminal residues of opd were replaced by the first 5 residues of lacZ resulted in the production of an active hydrolase identical in molecular mass to the hydrolase isolated from Flavobacterium sp. E. coli cells containing the lacZ-opd fusion showed higher levels of hydrolase activity than did cells containing the parent plasmid.
Mulbry
W W
WW
Pesticide Degradation Laboratory, U.S. Department of Agriculture, Beltsville, Maryland 20705.
Karns
J S
JS
D000595:Amino Acid Sequence; D043303:Aryldialkylphosphatase; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005417:Flavobacterium / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics* / Q000302:isolation & purification; D010957:Plasmids; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D015183:Restriction Mapping
D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; D010744:Phosphoric Monoester Hydrolases; D043303:Aryldialkylphosphatase