Uptake of the beta-lactam precursor alpha-aminoadipic acid in Penicillium chrysogenum is mediated by the acidic and the general amino acid permease.
External addition of the beta-lactam precursor alpha-aminoadipic acid to the filamentous fungus Penicillium chrysogenum leads to an increased intracellular alpha-aminoadipic acid concentration and an increase in penicillin production. The exact route for alpha-aminoadipic acid uptake is not known, although the general amino acid and acidic amino acid permeases have been implicated in this process. Their corresponding genes, PcGAP1 and PcDIP5, of P. chrysogenum were cloned and functionally expressed in a mutant of Saccharomyces cerevisiae (M4276) in which the acidic amino acid and general amino acid permease genes (DIP5 and GAP1, respectively) are disrupted. Transport assays show that both PcGap1 and PcDip5 mediated the uptake of alpha-aminoadipic acid, although PcGap1 showed a higher affinity for alpha-aminoadipic acid than PcDip5 (K(m) values, 230 and 800 microM, respectively). Leucine strongly inhibits alpha-aminoadipic acid transport via PcGap1 but not via PcDip5. This difference was exploited to estimate the relative contribution of each transport system to the alpha-aminoadipic acid flux in beta-lactam-producing P. chrysogenum. The transport measurements demonstrate that both PcGap1 and PcDip5 contribute to the alpha-aminoadipic acid flux.
Trip
Hein
H
Department of Microbiology, University of Groningen, Kerklaan 30, 9751 NN Haren, The Netherlands.
Evers
Melchior E
ME
Kiel
Jan A K W
JA
Driessen
Arnold J M
AJ
D015074:2-Aminoadipic Acid / Q000378:metabolism*; D000595:Amino Acid Sequence; D026905:Amino Acid Transport Systems / Q000378:metabolism*; D026922:Amino Acid Transport Systems, Acidic / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D008969:Molecular Sequence Data; D010408:Penicillium chrysogenum / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D026905:Amino Acid Transport Systems; D026922:Amino Acid Transport Systems, Acidic; D005656:Fungal Proteins; D011506:Proteins; D015074:2-Aminoadipic Acid