Significant advances have been made in the structure-based engineering of enzymes useful for beta-lactam antibiotic production. Structure-based engineering of penicillin G acylase and cephalosporin acylase has resulted in improved enzymes for use in enzymatic production processes. The structures of many other enzymes that could be used in the production of beta-lactam antibiotics, such as enzym...

a Department of Biochemistry and Molecular Biology I, Faculty of Biology, University 7 Complutense of Madrid, Madrid, Spain; and b Department of Environmental Biology, Centro 8 de Investigaciones Biológicas, CSIC, Madrid, Spain 9 10 Running head: Approach to catalytic mechanism of Penicillin V acylase from Streptomyces 11 lavendulae 12 #Address correspondence to Isabel de la Mata, [email protected]...

Synthesis of model peptide substrates and investigation of the reaction of their phenylacetyl protecting group enzyme transformation by means of penicillin G acylase D. L. Danalev*, L. K. Yotova 1 Department of Organic Chemistry, University of Chemical Technology and Metallurgy, 8 Kliment Ohridski Blvd., 1756 Sofia, Bulgaria 2 Department of Biotechnology, University of Chemical Technology and M...

Secretion of Escherichia coli penicillin acylase was improved by codon-based random mutagenesis of its signal peptide. The mutagenesis technology was applied to the gene region coding for positions Lys2 to Thr13 (N half) and Ala14 to Leu25 (C half) of the signal peptide. Protein secretion was higher in several signal peptide variants (up to fourfold with respect to the wild-type value).

The crystallization of three catalytically inactive mutants of penicillin V acylase (PVA) from Bacillus sphaericus in precursor and processed forms is reported. The mutant proteins crystallize in different primitive monoclinic space groups that are distinct from the crystal forms for the native enzyme. Directed mutants and clone constructs were designed to study the post-translational autoprote...

The penicillin G acylase genes from the Proteus rettgeri wild type and from a hyperproducing mutant which is resistant to succinate repression were cloned in Escherichia coli K-12. Expression of both wild-type and mutant P. rettgeri acylase genes in E. coli K-12 was independent of orientation in the cloning vehicle and apparently resulted from recognition in E. coli of the P. rettgeri promoter ...

A rapid and specific procedure was developed for the simultaneous detection of bacterial acylases and P-lactamases, using ampicillin and cephalexin as substrates. Bacterial suspensions from agar plates were incubated separately with each l-lactam substrate for 1 h at 37°C. The supernatant of the reaction mixture was dansylated, and the dansyl derivatives were separated by two-dimensional thin-l...