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Microbiology 142 (1996), 3219-3230
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microbiology, Vol 142, 3219-3230, Copyright © 1996 by Society for General Microbiology

ARTICLES

The purB gene of Escherichia coli K-12 is located in an operon

SM Green, T Malik, IG Giles and WT Drabble
Department of Biochemistry, University of Southampton, Crescent East, UK.

The structural gene (purB) for succinyl-AMP (S-AMP) lyase and three additional ORFs are on the same DNA strand of the chromosome of Escherichia coli. Cassette mutagenesis and primer extension mapping demonstrated that purB is co-transcribed with an upstream gene (ORF23, or ycfC) encoding a 22.9 kDa membrane-associated protein of non- essential, but unknown, function unrelated to purine biosynthesis. The purB operon lies between phoP and an ORF expressing an essential function which may correspond to asuE (trmU). S-AMP lyase was purified to near homogeneity. The purified enzyme is a homotetramer of 50 kDa subunits, has a K(m) for S-AMP of 3.7 microM and a pH optimum of 7.4- 7.6.


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