The crtS gene of Xanthophyllomyces dendrorhous encodes a novel cytochrome-P450 hydroxylase involved in the conversion of β-carotene into astaxanthin and other xanthophylls

Álvarez, Vanessa; Rodríguez‐Sáiz, Marta; Fuente, J. L. de la; Gudiña, Eduardo J.; Godio, Ramiro P.; Martı́n, Juan F.; Barredo, José Luis

doi:10.1016/j.fgb.2005.12.004

Cited by 90 publications

(72 citation statements)

References 38 publications

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“…Genes that specify polypeptides similar in amino acid sequence to HBFD are found The pathway in Paracoccus sp N81106 (previously known as Agrobacterium aurantiacum) was ascertained by Misawa et al (1995b). The pathway in X. dendrorhous (also known as Phaffia rhodozyma) has not been definitively established but is thought to operate using a single cytochrome P 450 enzyme (with a cytochrome P 450 reductase also needed to donate electrons; Alcaíno et al, 2008;Ukibe et al, 2009) to sequentially oxidize the number 3 and number 4 carbons of the two b-rings of b-carotene (Ojima et al, 2006;Á lvarez et al, 2006;Martín et al, 2008). Note that the biosynthesis of carotenoids with 3-hydroxy-4-keto-b-rings in the green alga H. pluvialis entails the use of an enzyme similar in amino acid sequence to the CrtW of Paracoccus sp N81106 to catalyze the addition of the carbonyl (Kajiwara et al, 1995;Lotan and Hirschberg, 1995), but hydroxylation of the number 3 carbon may be catalyzed by a cytochrome P 450 enzyme (Schoefs et al, 2001) rather than a CrtZ (or the related CHYb) enzyme.…”

Section: Online)mentioning

confidence: 99%

Elucidation of the Pathway to Astaxanthin in the Flowers of Adonis aestivalis

2011

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A few species in the genus Adonis are the only land plants known to produce the valuable red ketocarotenoid astaxanthin in abundance. Here, we ascertain the pathway that leads from the b-rings of b-carotene, a carotenoid ubiquitous in plants, to the 3-hydroxy-4-keto-b-rings of astaxanthin (3,39-dihydroxy-b,b-carotene-4,4'-dione) in the blood-red flowers of Adonis aestivalis, an ornamental and medicinal plant commonly known as summer pheasant's eye. Two gene products were found to catalyze three distinct reactions, with the first and third reactions of the pathway catalyzed by the same enzyme. The pathway commences with the activation of the number 4 carbon of a b-ring in a reaction catalyzed by a carotenoid b-ring 4-dehydrogenase (CBFD), continues with the further dehydrogenation of this carbon to yield a carbonyl in a reaction catalyzed by a carotenoid 4-hydroxy-b-ring 4-dehydrogenase, and concludes with the addition of an hydroxyl group at the number 3 carbon in a reaction catalyzed by the erstwhile CBFD enzyme. The A. aestivalis pathway is both portable and robust, functioning efficiently in a simple bacterial host. Our elucidation of the pathway to astaxanthin in A. aestivalis provides enabling technology for development of a biological production process and reveals the evolutionary origin of this unusual plant pathway, one unrelated to and distinctly different from those used by bacteria, green algae, and fungi to synthesize astaxanthin.

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Section: Online)mentioning

confidence: 99%

Elucidation of the Pathway to Astaxanthin in the Flowers of Adonis aestivalis

2011

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“…CrtS, is a 62.63 kDa (5.75 pI) fungal P450 that hydroxylates β-carotene to form astaxanthin. However, it is unrelated to the plant enzyme [70]. A P450 β-ring hydroxylase gene was also found in the carotenoid-containing thermophilic bacterium, Thermus thermophilus and the crystal structure has been solved [71].…”

Section: What Is the Structural Basis For Enzyme Specificity?mentioning

confidence: 99%

Escherichia coli as a platform for functional expression of plant P450 carotene hydroxylases

Quinlan

Jaradat

Wurtzel

2007

Archives of Biochemistry and Biophysics

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Carotenoids and their derivatives are essential for growth, development, and signaling in plants and have an added benefit as nutraceuticals in food crops. Despite the importance of the biosynthetic pathway, there remain open questions regarding some of the later enzymes in the pathway. The CYP97 family of P450 enzymes was predicted to function in carotene ring hydroxylation, to convert provitamin A carotenes to nonprovitamin A xanthophylls. However, substrate specificity was difficult to investigate directly in plants, which mask enzyme activities by a complex and dynamic metabolic network. To characterize the enzymes more directly, we amplified cDNAs from a model crop, Oryza sativa, and used functional complementation in Escherichia coli to test activity and specificity of members of Clans A and C. This heterologous system will be valuable for further study of enzyme interactions and substrate utilization needed to understand better the role of CYP97 hydroxylases in plant carotenoid biosynthesis.

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“…These findings suggest that only three genes are required for astaxanthin production from GGPP in X. dendrorhous. On the basis of the predicted protein sequence, crtS is presumed to encode a cytochrome P450 protein (3,22), and crtS-disrupted cells of X. dendrorhous do not synthesize astaxanthin. However, the CrtS protein has not been characterized, and its heterologous expression has been unsuccessful (17).…”

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Metabolic Engineering of Saccharomyces cerevisiae for Astaxanthin Production and Oxidative Stress Tolerance

Ukibe

Hashida

Yoshida

et al. 2009

Appl Environ Microbiol

133

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The red carotenoid astaxanthin possesses higher antioxidant activity than other carotenoids and has great commercial potential for use in the aquaculture, pharmaceutical, and food industries. In this study, we produced astaxanthin in the budding yeast Saccharomyces cerevisiae by introducing the genes involved in astaxanthin biosynthesis of carotenogenic microorganisms. In particular, expression of genes of the red yeast Xanthophyllomyces dendrorhous encoding phytoene desaturase (crtI product) and bifunctional phytoene synthase/lycopene cyclase (crtYB product) resulted in the accumulation of a small amount of ␤-carotene in S. cerevisiae. Overexpression of geranylgeranyl pyrophosphate (GGPP) synthase from S. cerevisiae (the BTS1 gene product) increased the intracellular ␤-carotene levels due to the accelerated conversion of farnesyl pyrophosphate to GGPP. Introduction of the X. dendrorhous crtS gene, encoding astaxanthin synthase, assumed to be the cytochrome P450 enzyme, did not lead to astaxanthin production. However, coexpression of CrtS with X. dendrorhous CrtR, a cytochrome P450 reductase, resulted in the accumulation of a small amount of astaxanthin. In addition, the ␤-carotene-producing yeast cells transformed by the bacterial genes crtW and crtZ, encoding ␤-carotene ketolase and hydroxylase, respectively, also accumulated astaxanthin and its intermediates, echinenone, canthaxanthin, and zeaxanthin. Interestingly, we found that these ketocarotenoids conferred oxidative stress tolerance on S. cerevisiae cells. This metabolic engineering has potential for overproduction of astaxanthin and breeding of novel oxidative stress-tolerant yeast strains.

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The crtS gene of Xanthophyllomyces dendrorhous encodes a novel cytochrome-P450 hydroxylase involved in the conversion of β-carotene into astaxanthin and other xanthophylls

Cited by 90 publications

References 38 publications

Elucidation of the Pathway to Astaxanthin in the Flowers of Adonis aestivalis

Elucidation of the Pathway to Astaxanthin in the Flowers of Adonis aestivalis

Escherichia coli as a platform for functional expression of plant P450 carotene hydroxylases

Metabolic Engineering of Saccharomyces cerevisiae for Astaxanthin Production and Oxidative Stress Tolerance

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