1. Penicillin G acylase from Achromobacter sp. CCM 4824 : an efficient biocatalyst for syntheses of beta-lactam antibiotics under conditions employed in large-scale processes
- Author
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Jaroslav Maršálek, Kamila Plhackova, Pavel Kyslík, Stanislav Bečka, Trupti K. Ashar, Rajasekar W. Vyasarayani, Marie Dobišová, Renáta Valešová, Michal Grulich, Martina Plačková, Andrea Palyzová, Anupama Datla, Václav Štěpánek, and Helena Marešová
- Subjects
medicine.drug_class ,Antibiotics ,Penicillanic Acid ,Achromobacter ,medicine.disease_cause ,beta-Lactams ,Applied Microbiology and Biotechnology ,Ampicillin ,Enzyme Stability ,medicine ,Escherichia coli ,Biotransformation ,chemistry.chemical_classification ,Chromatography ,Temperature ,Substrate (chemistry) ,Amoxicillin ,General Medicine ,Hydrogen-Ion Concentration ,Enzymes, Immobilized ,Anti-Bacterial Agents ,Penicillin ,Penicillin G Acylase ,Enzyme ,chemistry ,Penicillin Amidase ,Biotechnology ,medicine.drug - Abstract
Penicillin G acylase from Achromobacter sp. (NPGA) was studied in the enzymatic synthesis of β-lactam antibiotics by kinetically controlled N-acylation. When compared with penicillin acylase of Escherichia coli (PGA), the NPGA was significantly more efficient at syntheses of ampicillin and amoxicillin (higher S/H ratio and product accumulation) in the whole range of substrate concentrations. The degree of conversion of 6-aminopenicillanic acid to amoxicillin and ampicillin (160 mM 6-APA, 350 mM acyl donor methylester[Symbol: see text]HCl, pH 6.3, 25 °C, reaction time of 200 min) with immobilized NPGA equaled 96.9 % and 91.1 %, respectively. The enzyme was highly thermostable with maximum activity at 60 °C (pH 8.0) and 65 °C (pH 6.0). Activity half-life at 60 °C (pH 8.0) and at 60 °C (pH 6.0) was 24 min and 6.9 h, respectively. Immobilized NPGA exhibited long operational stability with half-life of about 2,000 cycles for synthesis of amoxicillin at conversion conditions used in large-scale processes (230 mM 6-APA, 340 mM D-4-hydroxyphenylglycine methylester[Symbol: see text]HCl, 27.5 °C, pH 6.25). We discuss our results with literature data available for related penicillin acylases in terms of their industrial potential.
- Published
- 2013