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Uptake of phenylacetic acid by two strains of Penicillium chrysogenum

✍ Scribed by Susanne Havn Eriksen; Tore Bengt Søderblom; Bo Jensen; Jørgen Olsen


Publisher
John Wiley and Sons
Year
1998
Tongue
English
Weight
105 KB
Volume
60
Category
Article
ISSN
0006-3592

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✦ Synopsis


Uptake of phenylacetic acid, the side-chain precursor of benzylpenicillin, was studied in Penicillium chrysogenum Wisconsin 54-1255 and in a strain yielding high levels of penicillin. In penicillin fermentations with the high-yielding strain, 100% recovery of phenylacetic acid in benzylpenicillin was found, whereas in the Wisconsin strain only 17% of the supplied phenylacetic acid was incorporated into benzylpenicillin while the rest was metabolized. Accumulation of total phenylacetic acidderived carbon in the cells was nonsaturable in both strains at high external concentrations of phenylacetic acid (250-3500 µM), and in the high-yielding strain at low phenylacetic acid concentrations (2.8-100 µM), indicating that phenylacetic acid enters the cells by simple diffusion, as concluded earlier for P. chrysogenum by other authors. However, at low external concentrations of phenylacetic acid saturable accumulation appeared in the Wisconsin strain. HPLC-analyses of cell extracts from the Wisconsin strain showed that phenylacetic acid was metabolized immediately after entry into the cells and different [ 14 C]-labeled metabolites were detected in the cells. Up to approximately 50% of the accumulated phenylacetic acid was metabolized during the transport-assay period, the conversion having an impact on the uptake experiments. Nevertheless, accumulation of free unchanged phenylacetic acid in the cells showed saturation kinetics, suggesting the possible involvement of a highaffinity carrier in uptake of phenylacetic acid in P. chrysogenum Wisconsin 54-1255. At high concentrations of phenylacetic acid, contribution to uptake by this carrier is minor in comparison to simple diffusion and therefore, of no importance in the industrial production of penicillin.


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