Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1994 Jun 1;300(Pt 2):355–358. doi: 10.1042/bj3000355

A semi-preparative enzymic synthesis of malonyl-CoA from [14C]acetate and 14CO2: labelling in the 1, 2 or 3 position.

G Roughan 1
PMCID: PMC1138169  PMID: 8002939

Abstract

A semi-preparative enzymic synthesis of [1-14C]malonyl-CoA from [1-14C]acetate and bicarbonate, and of [3-14C]malonyl-CoA from Na2(14)CO3 and acetate, was achieved by using chloroplasts rapidly isolated from 7-8-day-old pea shoots. Around 70% of the [1-14C]acetate was converted into malonyl-CoA in 2-3 h, and the specific radioactivity of [3-14C]malonyl-CoA synthesized in the system was 25-30 Ci/mol. Reactions were monitored and labelled products were purified by h.p.l.c.

Full text

PDF
355

Selected References

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

  1. Cerović Z. G., Plesnicar M. An improved procedure for the isolation of intact chloroplasts of high photosynthetic capacity. Biochem J. 1984 Oct 15;223(2):543–545. doi: 10.1042/bj2230543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Focke M., Feld A., Lichtenthaler K. Allicin, a naturally occurring antibiotic from garlic, specifically inhibits acetyl-CoA synthetase. FEBS Lett. 1990 Feb 12;261(1):106–108. doi: 10.1016/0014-5793(90)80647-2. [DOI] [PubMed] [Google Scholar]
  3. Gardiner S. E., Heinz E., Roughan P. G. Rates and products of long-chain Fatty Acid synthesis from [1-C]acetate in chloroplasts isolated from leaves of 16:3 and 18:3 plants. Plant Physiol. 1984 Apr;74(4):890–896. doi: 10.1104/pp.74.4.890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gornicki P., Haselkorn R. Wheat acetyl-CoA carboxylase. Plant Mol Biol. 1993 Jun;22(3):547–552. doi: 10.1007/BF00015984. [DOI] [PubMed] [Google Scholar]
  5. Hosokawa Y., Shimomura Y., Harris R. A., Ozawa T. Determination of short-chain acyl-coenzyme A esters by high-performance liquid chromatography. Anal Biochem. 1986 Feb 15;153(1):45–49. doi: 10.1016/0003-2697(86)90058-8. [DOI] [PubMed] [Google Scholar]
  6. Kannangara C. G., Jacobson B. S., Stumpf P. K. Fat Metabolism in Higher Plants: LVII. A Comparison of Fatty Acid-Synthesizing Enzymes in Chloroplasts Isolated from Mature and Immature Leaves of Spinach. Plant Physiol. 1973 Aug;52(2):156–161. doi: 10.1104/pp.52.2.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kim Y. S., Bang S. K. Malonyl coenzyme A synthetase. Purification and properties. J Biol Chem. 1985 Apr 25;260(8):5098–5104. [PubMed] [Google Scholar]
  8. Kim Y. S., Chae H. Z. Purification and properties of malonyl-CoA synthetase from Rhizobium japonicum. Biochem J. 1991 Feb 1;273(Pt 3):511–516. doi: 10.1042/bj2730511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mohan S. B., Kekwick R. G. Acetyl-coenzyme A carboxylase from avocado (Persea americana) plastids and spinach (Spinacia oleracea) chloroplasts. Biochem J. 1980 Jun 1;187(3):667–676. doi: 10.1042/bj1870667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Post-Beittenmiller D., Roughan G., Ohlrogge J. B. Regulation of plant Fatty Acid biosynthesis : analysis of acyl-coenzyme a and acyl-acyl carrier protein substrate pools in spinach and pea chloroplasts. Plant Physiol. 1992 Oct;100(2):923–930. doi: 10.1104/pp.100.2.923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Roughan G., Post-Beittenmiller D., Ohlrogge J., Browse J. Is Acetylcarnitine a Substrate for Fatty Acid Synthesis in Plants? Plant Physiol. 1993 Apr;101(4):1157–1162. doi: 10.1104/pp.101.4.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Roughan P. G., Holland R., Slack C. R. Acetate is the preferred substrate for long-chain fatty acid synthesis in isolated spinach chloroplasts. Biochem J. 1979 Dec 15;184(3):565–569. doi: 10.1042/bj1840565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Roughan P. G., Ohlrogge J. B. On the assay of acetyl-CoA synthetase activity in chloroplasts and leaf extracts. Anal Biochem. 1994 Jan;216(1):77–82. doi: 10.1006/abio.1994.1010. [DOI] [PubMed] [Google Scholar]
  14. Roughan P. G., Slack C. R., Holland R. High rates of [1-14C]acetate incorporation into the lipid of isolated spinach chloroplasts. Biochem J. 1976 Sep 15;158(3):593–601. doi: 10.1042/bj1580593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rutkoski A., Jaworski J. G. An improved synthesis of malonyl-coenzyme A. Anal Biochem. 1978 Nov;91(1):370–373. doi: 10.1016/0003-2697(78)90854-0. [DOI] [PubMed] [Google Scholar]
  16. Walker K. A., Harwood J. L. Localization of chloroplastic fatty acid synthesis de novo in the stroma. Biochem J. 1985 Mar 1;226(2):551–556. doi: 10.1042/bj2260551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wintermans J. F., de Mots A. Spectrophotometric characteristics of chlorophylls a and b and their pheophytins in ethanol. Biochim Biophys Acta. 1965 Nov 29;109(2):448–453. doi: 10.1016/0926-6585(65)90170-6. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES