Advertisement

Stereochemical Studies of Hydrogen Incorporation from Nucleotides with Fatty Acid Synthetase from Brevibacterium ammoniagenes

  • Y. Seyama
  • T. Kasama
  • T. Yamakawa
  • A. Kawaguchi
  • S. Okuda
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 101)

Summary

The biosynthesis of fatty acids from malonyl-CoA and acetyl-CoA was investigated with an enzyme preparation which was purified 100-fold from Brevibacterium ammoniagenes. Fatty acids synthesized in the presence of D2O and stereospecifically deuterated NADPH and NADH were isolated and analyzed by mass chromatography to examine the localization of deuterium in the molecule. The following results were obtained: 1) HB hydrogen of NADPH was used for (ß-ketoacyl reductase. 2) HB hydrogen of NADH was used for enoyl reductase. 3) Hydrogen atoms from water were found on the even-numbered methylene carbon atoms (2-hydrogen atoms per carbon atom) and some were also found on the odd-numbered methylene carbon. 4) Hydrogen atoms from NADPH were found on odd-numbered methylene carbon atoms (1-hydrogen per carbon). 5) Hydrogen atoms from NADH were also found on the odd-numbered methylene carbon atoms, but the number of incorporated hydrogen atoms was less than expected. The exchange of HB hydrogen of NADH with water catalyzed by enoyl reductase was suspected. 6) The exchange of methylene hydrogen atoms of malonyl-CoA with proton of water was suggested by 13C NMR analysis.

Keywords

Methyl Oleate Malonic Acid Hydrogen Exchange Deuterium Atom Methylene Carbon 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Reference

  1. 1.
    Kawaguchi,A.& Okuda,S.(1977) PrOC.Natl.Acad.SCi.U.S.74,3180–3183 Google Scholar
  2. 2.
    Foster,D.W.& Bloom,B. (1963) J.Biol.Chem., 238, 888–892PubMedGoogle Scholar
  3. 3.
    Foster,D.W.& Katz,J. (1966) Biochim.Biophys.Acta, 125, 422–427PubMedCrossRefGoogle Scholar
  4. 4.
    Jungus,R.L. (1968) Biochemistry, 7, 3708–3717CrossRefGoogle Scholar
  5. 5.
    Cornforth,J.W.,Cornforth,R.H.,Donninger,C.,Popjak,G.Ryback,G.& Schoepfer,G.J. (1965) Proc.Roy.Soc.B., 163, 436–464CrossRefGoogle Scholar
  6. 6.
    Colowick,S.P.& Kaplan,N.O.(1957) in Methods in Enzymology IV,840–848Google Scholar
  7. 7.
    Ryhage,R.& Stenhagen,E.(1963) in Mass Spectrometry of Organic Ions (McLafferty,F.W.,ed.) Chapt.9, Academic Press,New YorkGoogle Scholar
  8. 8.
    Drysdale,G.R. (1959) J.Biol.Chem. 234, 2399–2403PubMedGoogle Scholar
  9. 9.
    Gansow,O.A.& Schittenhelm,W. (1971) J.Amer.Chem.Soc., 93, 4294–4295CrossRefGoogle Scholar
  10. 10.
    Arnstadt,K.-I.,Schindlbeck,G.,& Lynen,F. (1975) Eur.J.Biochem. 55, 561–571PubMedCrossRefGoogle Scholar
  11. 11.
    Foster,D.W.& Bloom,B. (1962) Biochim.Biophys.Acta, 6O, 189–190CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1978

Authors and Affiliations

  • Y. Seyama
    • 1
  • T. Kasama
    • 1
  • T. Yamakawa
    • 1
  • A. Kawaguchi
    • 2
  • S. Okuda
    • 2
  1. 1.Dept. of Biochem., Faculty of MedicineUniversity of TokyoTokyoJapan
  2. 2.Institute of Applied MicrobiologyUniversity of TokyoTokyoJapan

Personalised recommendations