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Parathion Hydrolase Gene from Pseudomonas diminuta MG: Subcloning, Complete Nucleotide Sequence, and Expression of the Mature Portion of the Enzyme in Escherichia coli

Bio/Technology volume 7, pages 1151–1155 (1989)Cite this article

Abstract

The nucleotide sequence of the gene from Pseudomonas diminuta MG encoding parathion hydrolase was determined, and a single open reading frame located. Comparison of the deduced N-terminal amino acid sequence with that determined by protein sequence analysis of the enzyme from P. diminuta MG indicated that parathion hydrolase is synthesized as a 365 amino acid precursor from which 29 amino acids are removed. Expression of the processed parathion hydrolase coding sequence in Escherichia coli under the control of lambda PL promoter results in the production of high-level enzyme activity. Furthermore, addition of metal salts to the growth medium enhanced specific activity. The N-terminal amino acid sequence, C-terminal amino acid sequence and the amino acid composition of the purified enzyme were in agreement to those expected from the translated DNA sequence.

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Authors and Affiliations

  1. Amgen Inc., Thousand Oaks, CA, 91320

    Cüneyt M. Serdar, Douglas C. Murdock & Michael F. Rohde

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  1. Cüneyt M. Serdar
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Serdar, C., Murdock, D. & Rohde, M. Parathion Hydrolase Gene from Pseudomonas diminuta MG: Subcloning, Complete Nucleotide Sequence, and Expression of the Mature Portion of the Enzyme in Escherichia coli. Nat Biotechnol 7, 1151–1155 (1989). https://doi.org/10.1038/nbt1189-1151

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