| Products/Services Used | Details | Operation |
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| Recombinant Proteins | The selected beta-lactamases were optimized for protein expression in Escherichia coliand synthesized by GenScript (Piscataway, NJ, USA) and then cloned into the pET24a(+) expression vector. Recombinant β-lactamases were expressed in E. coli BL21(DE3)-pGro7/GroEL (TaKaRa) using ZYP-5052 media. Each culture was grown at 37°C until reaching an OD600 nm = 0.8, followed by addition of L-arabinose (0.2% m/v) and induction with a temperature transition to 16°C over 20 hours. | Get A Quote |
β-lactams targeting the bacterial cell wall are not efficient on archaea. Using phylogenetic analysis and common ancestor sequences for bacterial β-lactamases, we found serendipitously class B and class C-like β-lactamase genes in most archaea genomes. The class B β-lactamase appears to be highly conserved in archaea and to has been transferred in the bacterial genus Elizabethkingia. The experimentaly expressed class B enzyme from Methanosarcina barkeri was able to digest penicillin G and was inhibited by a β-lactamase inhibitor (i.e. sulbactam). The class C-like β-lactamase was more closely related to DD-peptidase enzymes than know bacterial class C β-lactamases. The use of these very conserved genes... More
