Physiological characterisation of Penicillium chrysogenum strains expressing the expandase gene from Streptomyces clavuligerus during batch cultivations. Growth and adipoyl-7-aminodeacetoxycephalosporanic acid production.
The production of adipoyl-7-aminodeacetoxy-cephalosporanic acid (ad-7-ADCA) was studied, using two recombinant strains of Penicillium chrysogenum carrying the expandase gene from Streptomyces clavuligerus. The adipoyl-side chain of this compound may easily be removed using an amidase; and this process therefore represents a new route for the production of 7-ADCA, which serves as a precursor for the production of many semi-synthetic cephalosporins. In this study, one low- and one high-yielding strains were characterised and the specific productivities of ad-7-ADCA and byproducts of the biosynthetic pathway were compared. The fluxes through the biosynthetic pathway were quantified and it was found that there was a 30% higher flux through the expandase in the high-yielding strain. In both strains, there was a significant degradation of adipate. Furthermore, the initial adipate concentration in batch cultures was shown to have a positive effect on the formation of ad-7-ADCA.
Robin
J
J
Centre for Process Biotechnology, Biocentrum-DTU, Technical University of Denmark, Lyngby.
Jakobsen
M
M
Beyer
M
M
Noorman
H
H
Nielsen
J
J
D000272:Adipates / Q000378:metabolism; D002511:Cephalosporins / Q000032:analysis / Q000378:metabolism*; D005632:Fructose / Q000378:metabolism; D005942:Gluconates / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D019751:Intramolecular Transferases / Q000235:genetics / Q000378:metabolism*; D016254:Mutagenesis, Insertional; D010397:Penicillanic Acid / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism; D046915:Penicillin-Binding Proteins; D010408:Penicillium chrysogenum / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D013302:Streptomyces / Q000235:genetics; D013395:Sucrose / Q000378:metabolism
D000272:Adipates; D002511:Cephalosporins; D005942:Gluconates; D046915:Penicillin-Binding Proteins; C424182:adipoyl-7-amino-3-deacetoxycephalosporanic acid; D005632:Fructose; D013395:Sucrose; C029900:adipic acid; D010397:Penicillanic Acid; D019751:Intramolecular Transferases; C038371:deacetoxycephalosporin C synthetase; D005947:Glucose; C004408:aminopenicillanic acid; C030691:gluconic acid