title: Penicillin G acylase from Achromobacter sp. CCM 4824 : an efficient biocatalyst for syntheses of beta-lactam antibiotics under conditions employed in large-scale processes.
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.
authors.affiliation: Laboratory of Enzyme Technology, Institute of Microbiology ASCR, v.v.i., Prague, Czech Republic.
authors.forename: Stanislav
authors.initials: S
authors.lastname: Bečka
authors.forename: Václav
authors.initials: V
authors.lastname: Štěpánek
authors.forename: Rajasekar W
authors.initials: RW
authors.lastname: Vyasarayani
authors.forename: Michal
authors.initials: M
authors.lastname: Grulich
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Maršálek
authors.forename: Kamila
authors.initials: K
authors.lastname: Plháčková
authors.forename: Marie
authors.initials: M
authors.lastname: Dobišová
authors.forename: Helena
authors.initials: H
authors.lastname: Marešová
authors.forename: Martina
authors.initials: M
authors.lastname: Plačková
authors.forename: Renáta
authors.initials: R
authors.lastname: Valešová
authors.forename: Andrea
authors.initials: A
authors.lastname: Palyzová
authors.forename: Anupama
authors.initials: A
authors.lastname: Datla
authors.forename: Trupti K
authors.initials: TK
authors.lastname: Ashar
authors.forename: Pavel
authors.initials: P
authors.lastname: Kyslík
mesh_terms: D042402:Achromobacter / Q000201:enzymology*; D000658:Amoxicillin / Q000378:metabolism; D000667:Ampicillin / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D001711:Biotransformation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D010397:Penicillanic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D010399:Penicillin Amidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D013696:Temperature; D047090:beta-Lactams / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D004800:Enzymes, Immobilized; D047090:beta-Lactams; D000667:Ampicillin; D000658:Amoxicillin; D010397:Penicillanic Acid; D010399:Penicillin Amidase; C004408:aminopenicillanic acid