Skip to main content
PMC home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1975 Sep;8(3):321–327. doi: 10.1128/aac.8.3.321

Biosynthesis of Mycophenolic Acid: Purification and Characterization of S-Adenosyl-l-Methionine:Demethylmycophenolic Acid O-Methyltransferase

W L Muth a, C H Nash III a,1
PMCID: PMC429313  PMID: 241289

Abstract

The final step in the biosynthesis of mycophenolic acid involves the transfer of a methyl group from S-adenosylmethionine to demethylmycophenolic acid. The enzyme, S-adenosylmethionine:demethylmycophenolic acid O-methyltransferase, was isolated from Penicillium stoloniferum and purified 2,700-fold by ammonium sulfate fractionation and diethylaminoethyl-cellulose and Sephadex G-200 chromatography. Maximum enzyme activity was achieved at pH 7.5 and a temperature of 27 to 28 C. The apparent Km for demethylmycophenolic acid was 3.1 × 10−6 M. The enzyme preparation was 50% inactivated when exposed to 33 C for 15 min. Mycophenolic acid, homocystine, S-adenosyl-homocysteine, ethanol, and Mg2+ inhibited the methyltransferase. This enzyme appears to be subject to end product inhibition which may regulate the synthesis of mycophenolic acid. The methyltransferase activity was highest during the early phases of the fermentation.

Full text

PDF
325

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abraham E. P. The effect of mycophenolic acid on the growth of Staphylococcus aureus in heart broth. Biochem J. 1945;39(5):398–408. doi: 10.1042/bj0390398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ando K., Suzuki S., Tamura G., Arima K. Antiviral activity of mycophenolic acid. Studies on antiviral and antitumor antibiotics. IV. J Antibiot (Tokyo) 1968 Nov;21(11):649–652. doi: 10.7164/antibiotics.21.649. [DOI] [PubMed] [Google Scholar]
  3. Canonica L., Kroszczynski W., Ranzi B. M., Rindone B., Santaniello E., Scolastico C. Biosynthesis of mycophenolic acid. J Chem Soc Perkin 1. 1972;21:2639–2643. doi: 10.1039/p19720002639. [DOI] [PubMed] [Google Scholar]
  4. Clutterbuck P. W., Oxford A. E., Raistrick H., Smith G. Studies in the biochemistry of micro-organisms: The metabolic products of the Penicillium brevi-compactum series. Biochem J. 1932;26(5):1441–1458. doi: 10.1042/bj0261441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. JAUREGUIBERRY G., FARRAGIA-FOUGEROUSE G., AUDIER H., LEDERER E. SUR LE TRANSFERT DU GROUPEMENT -CD-3 DE LA M'ETHIONINE-CD-3 AU COURS DE LA BIOSYNTH'ESE DE L'ACIDE MYCOPH'ENOLIQUE. C R Hebd Seances Acad Sci. 1964 Nov 2;259:3108–3110. [PubMed] [Google Scholar]
  6. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  7. Miller P. A., Saturnelli A., Martin J. H., Itscher L. A., Bohonos N. A new family of tetracycline precursors. N-demethylanhydrotetracyclines. Biochem Biophys Res Commun. 1964 Jun 15;16(3):285–291. doi: 10.1016/0006-291x(64)90341-9. [DOI] [PubMed] [Google Scholar]
  8. Mitsui A., Suzuki S. Immunosuppressive effect of mycophenolic acid. J Antibiot (Tokyo) 1969 Aug;22(8):358–363. doi: 10.7164/antibiotics.22.358. [DOI] [PubMed] [Google Scholar]
  9. Noto T., Sawada M., Ando K., Koyama K. Some biological properties of mycophenolic acid. J Antibiot (Tokyo) 1969 Apr;22(4):165–169. doi: 10.7164/antibiotics.22.165. [DOI] [PubMed] [Google Scholar]
  10. Rao M. M., Rebello P. F., Pogell B. M. Biosynthesis of puromycin in Streptomyces alboniger. Enzymatic methylation of O-demethylpuromycin. J Biol Chem. 1969 Jan 10;244(1):112–118. [PubMed] [Google Scholar]
  11. Sachs D. H., Painter E. Improved flow rates with porous sephadex gels. Science. 1972 Feb 18;175(4023):781–782. doi: 10.1126/science.175.4023.781. [DOI] [PubMed] [Google Scholar]
  12. Sankaran L., Pogell B. M. A simple, one-step assay for methyltransferase activity: direct partitioning of radioactive product into scintillation fluid. Anal Biochem. 1973 Jul;54(1):146–152. doi: 10.1016/0003-2697(73)90257-1. [DOI] [PubMed] [Google Scholar]
  13. Williams R. H., Lively D. H., DeLong D. C., Cline J. C., Sweeny M. J. Mycophenolic acid: antiviral and antitumor properties. J Antibiot (Tokyo) 1968 Jul;21(7):463–464. doi: 10.7164/antibiotics.21.463. [DOI] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES