Biotechnology Letters

, Volume 26, Issue 10, pp 813–817 | Cite as

Cloning of two carotenoid ketolase genes from Nostoc punctiforme for the heterologous production of canthaxanthin and astaxanthin

  • Sabine Steiger
  • Gerhard Sandmann


For the heterologous synthesis of keto-carotenoids such as astaxanthin, two carotenoid ketolase genes crtW38 and crtW148, were cloned from the cyanobacterium, Nostoc punctiforme PCC 73102 and functionally characterized. Upon expression in Escherichia coli, both genes mediated the conversion of β-carotene to canthaxanthin. However in a zeaxanthin-producing E. coli, only the gene product of crtW148 introduced 4-keto groups into the 3,3′-dihydroxy carotenoid zeaxanthin yielding astaxanthin. The gene product of crtW38 was unable to catalyze this reaction. Both ketolases differ in their interaction with a hydroxylase in the biosynthetic pathway from β-carotene to astaxanthin.

astaxanthin canthaxanthin β-carotene ketolase heterologous carotenoid production 


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  1. Albrecht M, Steiger S, Sandmann G (2001) Expression of a ketolase gene mediates the synthesis of canthaxanthin in Synechococcus leading to resistance against pigment photodegradation and UVB sensitivity of photosynthesis. Photochem. Photobiol. 73: 551–555.Google Scholar
  2. Borovkov AY, Rivkin MI (1997) pBBR1MCS: XcmI-containing vector for direct cloning of pcr products. BioTechniques 22: 812–814.Google Scholar
  3. Breitenbach J, Misawa N, Kajiwara S, Sandmann G (1996) Expression in Escherichia coli and properties of the carotene ketolase from Haematococcus pluvialis. FEMS Microbiol. Lett. 140: 241–246.Google Scholar
  4. Fernández-Gonzalez B, Sandmann G, Vioque A (1997) A new type of asymmetrically acting â-carotene ketolase is required for the synthesis of echinenone in the cyanobacterium Synechocystis sp. PCC 6803. J. Biol. Chem. 272: 9728–9733.Google Scholar
  5. Fraser PD, Shimada H, Misawa N (1998) Enzymic confirmation of reactions involved in routes to astaxanthin formation, elucidated using a direct substrate in vitro assay. Eur. J. Biochem. 252: 229–236.Google Scholar
  6. Goodwin TW (1980) The Biochemistry of the Carotenoids, Vol. 1: Plants, 2nd edn. New York: Chapman & Hall.Google Scholar
  7. Harker M, Hirschberg J (1999) Carotenoid biosynthesis genes in the bacterium Paracoccus marcusii MH1. Submission to data base: Accession No. Y15112.Google Scholar
  8. Johnson EA, An GH (1991) Astaxanthin from microbial sources. Crit. Rev. Biotechnol. 11: 297–326.Google Scholar
  9. Lorenz RT, Cysewski GR (2000) Commercial potential for Haematococcus microalgae as a natural source of astaxanthin. Trends Biotechnol. 18: 160–167.Google Scholar
  10. Meyers SP (1994) Developments in world aquaculture, feed formulations and role of carotenoids. Pure Appl. Chem. 66: 1069–1076.Google Scholar
  11. Misawa N, Satomi Y, Kondo K, Yokoyama A, Kajiwara S, Saito T, Ohtani T, Miki W (1995) Structure and functional analysis of a marine bacterial carotenoid biosynthesis gene cluster and astaxanthin biosynthetic pathway proposed at the gene level. J. Bacteriol. 177: 6575–6584.Google Scholar
  12. Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J. Gen. Microbiol. 111: 1–61.Google Scholar
  13. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.Google Scholar
  14. Verdoes J, Krubasik P, Sandmann G, van Ooyen M (1999) Isolation and functional characterization of a novel type of carotenoid biosynthetic gene from Xanthophyllomyces dendrorhous. Mol. Gen. Genet. 262: 453–461.Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Sabine Steiger
    • 1
  • Gerhard Sandmann
    • 1
  1. 1.Biosynthesis Group, Botanical InstituteGoethe UniversityFrankfurtGermany

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