Cloning, sequencing, and enhanced expression of the dihydropteroate synthase gene of Escherichia coli MC4100
- 1 September 1992
- journal article
- research article
- Published by American Society for Microbiology in Journal of Bacteriology
- Vol. 174 (18), 5961-5970
- https://doi.org/10.1128/jb.174.18.5961-5970.1992
Abstract
The Escherichia coli gene coding for dihydropteroate synthase (DHPS) has been cloned and sequenced. The protein has 282 amino acids and a compositional molecular mass of 30,314 daltons. Increased expression of the enzyme was realized by using a T7 expression system. The enzyme was purified and crystallized. A temperature-sensitive mutant was isolated and found to express a DHPS with a lower specific activity and lower affinities for para-aminobenzoic acid and sulfathiazole. The allele had a point mutation that changed a phenylalanine codon to a leucine codon, and the mutation was in a codon that is conserved among published DHPS sequences.Keywords
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