Subcellular compartmentation of penicillin biosynthesis in Penicillium chrysogenum. The amino acid precursors are derived from the vacuole.
The cellular localization of the origin of alpha-aminoadipate used in penicillin biosynthesis and the first enzymic step in Penicillium chrysogenum involved, delta-(alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase (ACVS), has been studied. Subcellular fractions were obtained from protoplasts of a high penicillin-producing strain upon lysis by Triton X-100, and vacuoles purified from them. They were identified by the aid of alpha-mannosidase as a marker enzyme, by the presence of polyphosphate, and their ability to sequester [14C]lysin, added to the protoplasts prior to subcellular fractionation. 15.6 and 26.5%, respectively, of 6-[14C]alpha-aminoadipate, and 8.5 and 10.3%, respectively, of [14C]valine added accordingly were also found in the vacuole, and the higher proportion was found in vacuoles isolated from penicillin-producing mycelia. ACVS protein was detected in the membrane as well as the soluble fraction of the purified vacuoles. We propose therefore that ACVS is located either within or bound to the vacuolar membrane, and that the precursor amino acids for penicillin biosynthesis are withdrawn from the vacuolar amino acid pool.
Lendenfeld
T
T
Abteilung für Mikrobielle Biochemie, Technische Universität Wien, Vienna, Austria.
Ghali
D
D
Wolschek
M
M
Kubicek-Pranz
E M
EM
Kubicek
C P
CP
D015074:2-Aminoadipic Acid / Q000096:biosynthesis*; D000596:Amino Acids / Q000378:metabolism*; D002474:Cell-Free System; D008239:Lysine / Q000378:metabolism; D008954:Models, Biological; D010406:Penicillins / Q000096:biosynthesis*; D010408:Penicillium chrysogenum / Q000378:metabolism*; D010453:Peptide Synthases / Q000378:metabolism; D011122:Polyphosphates / Q000378:metabolism; D011523:Protoplasts / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism*; D014633:Valine / Q000378:metabolism
D000596:Amino Acids; D010406:Penicillins; D011122:Polyphosphates; D015074:2-Aminoadipic Acid; D010453:Peptide Synthases; C095368:alpha-aminoadipyl-cysteinyl-valine synthetase; D014633:Valine; D008239:Lysine