Design of an enzymatic hybrid system: a useful strategy for the biosynthesis of benzylpenicillin in vitro.
A hybrid (prokaryotic-eukaryotic) enzyme system leading to the production of benzylpenicillin has been developed. In vitro synthesis of penicillin G was achieved by incubating 6-aminopenicillanic acid, CoA, phenylacetic acid, homogeneously pure phenylacetyl-CoA ligase (PA-CoA ligase) from Pseudomonas putida and acyl-CoA:6-APA acyltransferase (AT) from Penicillium chrysogenum. Benzylpenicillin was also obtained when AT was coupled with PA-CoA ligase and isopenicillin N-synthetase (IPNS). This is the first description of an in vitro assay that, using enzymes of different microbial origin, mimics the three last enzymatic steps leading to the biosynthesis of penicillin G in P. chrysogenum.
Martínez-Blanco
H
H
Departamento de Bioquímica y Biología Molecular, Facultad de Veterinaria, Universidad de León, Spain.
Reglero
A
A
Martín-Villacorta
J
J
Luengo
J M
JM
D000217:Acyltransferases / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003065:Coenzyme A / Q000378:metabolism; D003066:Coenzyme A Ligases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010088:Oxidoreductases / Q000378:metabolism; D010397:Penicillanic Acid / Q000378:metabolism / Q000031:analogs & derivatives; D010400:Penicillin G / Q000378:metabolism*; D046915:Penicillin-Binding Proteins; D010408:Penicillium chrysogenum / Q000201:enzymology*; D010648:Phenylacetates / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology*; D013696:Temperature
D046915:Penicillin-Binding Proteins; D010648:Phenylacetates; D010397:Penicillanic Acid; D010088:Oxidoreductases; C038370:isopenicillin N synthetase; D000217:Acyltransferases; C018756:acyl-CoA-6-aminopenicillanic acid acyltransferase; D003066:Coenzyme A Ligases; C023914:phenylacetate - CoA ligase; C025136:phenylacetic acid; D010400:Penicillin G; C004408:aminopenicillanic acid; D003065:Coenzyme A