A new screening method for bacteria capable of producing penicillin acylase is described. The method is based on the use of Serratia marcescens sensitive to 6-aminopenicillanic acid but comparatively resistant to benzylpenicillin. It is simple, quite specific, and requires no special equipment. It can also be used to screen for phenoxymethylpenicillin acylase activity. We also suggest an acidim...

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...

BACKGROUND Immobilized Penicillin G Acylase (PGA) derivatives are biocatalysts that are industrially used for the hydrolysis of Penicillin G by fermentation and for the kinetically controlled synthesis of semi-synthetic beta-lactam antibiotics. One of the most used supports for immobilization is glyoxyl-activated agarose, which binds the protein by reacting through its superficial Lys residues....

Aculeacin A acylase from Actinoplanes utahensis produced by Streptomyces lividans revealed acylase activities that are able to hydrolyze penicillin V and several natural aliphatic penicillins. Penicillin K was the best substrate, showing a catalytic efficiency of 34.79 mM(-1) s(-1). Furthermore, aculeacin A acylase was highly thermostable, with a midpoint transition temperature of 81.5 degrees C.

The large heterodimeric penicillin G acylase from Alcaligenes faecalis was displayed on the surface of phage fd. We fused the coding sequence (alpha subunit-internal peptide-beta subunit) to the gene of a phage coat protein. A modified g3p signal sequence was used to direct the polypeptide to the periplasm. Here we show that a heterodimeric enzyme can be expressed as a fusion protein that matur...

This study is pertaining to the production of penicillin G acylase (PGA) by Bacillus subtilis 8105MU331 in which PGA gene, under the control of thermal-induced promoter, was integrated. The key process parameters including induced-temperature, induced-time, and culture temperature were optimized in flask culture. A three-stage cultivation process was developed for PGA production with the expres...

The enzymatic synthesis of amoxicillin is catalyzed by Penicillin G Acylase (PGA). As byproducts, hydroxyphenylglycine and alcohol are also formed from hydrolytic reactions and antibiotic synthesis, respectively. The design of this process should be directed to promote the synthesis reaction. At the same time, it is necessary to reduce the hydrolytic reaction of amoxicillin through its crystall...

We used ultraviolet (UV) radiation to induce mutation in three locally isolated strains of Escherichia coli. Different dilutions of bacterial cultures were exposed to UV lamp of 254 nm wavelength for different time intervals at varied distances ranging from 5 to 210 sec and 5 to 100 cm. Viable colonies were screened for mutants with an increased production of penicillin G acylase (PGA) and a re...

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...