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Expression of Xanthophyllomyces dendrorhous cytochrome-P450 hydroxylase and reductase in Mucor circinelloides

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Abstract

Carotenoids are natural pigments that act as powerful antioxidants and have various beneficial effects on human and animal health. Mucor circinelloides (Mucoromycotina) is a carotenoid producing zygomycetes fungus, which accumulates β-carotene as the main carotenoid but also able to produce the hydroxylated derivatives of β-carotene (i.e. zeaxanthin and β-cryptoxanthin) in low amount. These xanthophylls, together with the ketolated derivatives of β-carotene (such as canthaxanthin, echinenone and astaxanthin) have better antioxidant activity than β-carotene. In this study our aim was to modify and enhance the xanthophyll production of the M. circinelloides by expression of heterologous genes responsible for the astaxanthin biosynthesis. The crtS and crtR genes, encoding the cytochrome-P450 hydroxylase and reductase, respectively, of wild-type and astaxanthin overproducing mutant Xanthophyllomyces dendrorhous strains were amplified from cDNA and the nucleotide and the deduced amino acid sequences were compared to each other. Introduction of the crtS on autonomously replicating plasmid in the wild-type M. circinelloides resulted enhanced zeaxanthin and β-cryptoxanthin accumulation and the presence of canthaxanthin, echinenone and astaxanthin in low amount; the β-carotene hydroxylase and ketolase activity of the X. dendrorhous cytochrome-P450 hydroxylase in M. circinelloides was verified. Increased canthaxanthin and echinenone production was observed by expression of the gene in a canthaxanthin producing mutant M. circinelloides. Co-expression of the crtR and crtS genes led to increase in the total carotenoid and slight change in xanthophyll accumulation in comparison with transformants harbouring the single crtS gene.

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Acknowledgments

The research of Árpád Csernetics and Csaba Vágvölgyi was supported by the European Union and the State of Hungary, co-financed by the European Social Fund in the framework of TÁMOP-4.2.4.A/2-11/1-2012-0001 ‘National Excellence Program’. The infrastructure and research equipment was supported by TÁMOP-4.1.1.C-12/1/KONV-2012-0014 and OTKA NN 106394.

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Authors and Affiliations

  1. Department of Microbiology, Faculty of Science and Informatics, University of Szeged, Közép fasor 52., Szeged, 6726, Hungary

    Árpád Csernetics, Eszter Tóth, Anita Farkas, Gábor Nagy, Ottó Bencsik, Csaba Vágvölgyi & Tamás Papp

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  1. Árpád Csernetics
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  2. Eszter Tóth
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  3. Anita Farkas
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  4. Gábor Nagy
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  5. Ottó Bencsik
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  7. Tamás Papp
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Correspondence to Árpád Csernetics.

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Csernetics, Á., Tóth, E., Farkas, A. et al. Expression of Xanthophyllomyces dendrorhous cytochrome-P450 hydroxylase and reductase in Mucor circinelloides . World J Microbiol Biotechnol 31, 321–336 (2015). https://doi.org/10.1007/s11274-014-1784-z

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