PMID:16085816

Citation	Nishida, Y, Adachi, K, Kasai, H, Shizuri, Y, Shindo, K, Sawabe, A, Komemushi, S, Miki, W and Misawa, N (2005) Elucidation of a carotenoid biosynthesis gene cluster encoding a novel enzyme, 2,2'-beta-hydroxylase, from Brevundimonas sp. strain SD212 and combinatorial biosynthesis of new or rare xanthophylls. Appl. Environ. Microbiol. 71:4286-96
Abstract	A carotenoid biosynthesis gene cluster mediating the production of 2-hydroxyastaxanthin was isolated from the marine bacterium Brevundimonas sp. strain SD212 by using a common crtI sequence as the probe DNA. A sequence analysis revealed this cluster to contain 12 open reading frames (ORFs), including the 7 known genes, crtW, crtY, crtI, crtB, crtE, idi, and crtZ. The individual ORFs were functionally analyzed by complementation studies using Escherichia coli that accumulated various carotenoid precursors due to the presence of other bacterial crt genes. In addition to functionally identifying the known crt genes, we found that one (ORF11, named crtG) coded for a novel enzyme, carotenoid 2,2'-beta-hydroxylase, which showed intriguingly partial homology with animal sterol-C5-desaturase. When this crtG gene was introduced into E. coli accumulating zeaxanthin and canthaxanthin, the resulting transformants produced their 2-hydroxylated and 2,2'-dihydroxylated products which were structurally novel or rare xanthophylls, as determined by their nuclear magnetic resonance and high-performance liquid chromatography/photodiode array detector/atmospheric pressure chemical ionization mass spectrometry spectral data. The new carotenoid produced was suggested to have a strong inhibitory effect on lipid peroxidation.
Links	PubMed PMC1183362 Online version:10.1128/AEM.71.8.4286-4296.2005
Keywords	Amino Acid Sequence; Animals; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Biotechnology/methods; Carotenoids/biosynthesis; Carotenoids/chemistry; Carotenoids/genetics; Caulobacteraceae/enzymology; Caulobacteraceae/genetics; Caulobacteraceae/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli/metabolism; Humans; Mice; Mixed Function Oxygenases/genetics; Mixed Function Oxygenases/metabolism; Molecular Sequence Data; Multigene Family; Xanthophylls/biosynthesis
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
9CAUL:Q4W8B6	involved_in	GO:0016117: carotenoid biosynthetic process	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
9CAUL:Q4W8B6		GO:0016117: carotenoid biosynthetic process	ECO:0000314:		P		The sequence of the cartenoid biosynthesis gene cluster is shown in figure 1a. 1b shows the alignment of the region with that of other species. Gene from this region were inserted into E. coli and tagged with a specific lead sequence that makes them more efficiently translated. Table 3 shows the identification by spectroscopy of cartenoid precursors in a number of transformants expressing different combinations of crt genes.	complete CACAO 2965
9CAUL:Q4W8B5	involved_in	GO:0016117: carotenoid biosynthetic process	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
9CAUL:Q4W8B5		GO:0016117: carotenoid biosynthetic process	ECO:0000314:		P		The sequence of the cartenoid biosynthesis gene cluster is shown in figure 1a. 1b shows the alignment of the region with that of other species. Genes from this region were inserted into E. coli and tagged with a specific lead sequence that makes them more efficiently translated. Table 3 shows the identification by spectroscopy of cartenoid precursors in a number of transformants expressing different combinations of crt genes.	complete CACAO 2966
9CAUL:Q4W8B7		GO:0016117: carotenoid biosynthetic process	ECO:0000314:		P		Table 3 shows the identification by spectroscopy of cartenoid precursors in a number of transformants expressing different combinations of crt genes.	complete CACAO 2967
9CAUL:Q4W8B7	involved_in	GO:0016117: carotenoid biosynthetic process	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
9CAUL:Q4W8B2	involved_in	GO:0016117: carotenoid biosynthetic process	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
9CAUL:Q4W8B2		GO:0016117: carotenoid biosynthetic process	ECO:0000314:		P		The sequence of the cartenoid biosynthesis gene cluster is shown in figure 1a. 1b shows the alignment of the region with that of other species. Genes from this region were inserted into E. coli and tagged with a specific lead sequence that makes them more efficiently translated. Table 3 shows the identification by spectroscopy of cartenoid precursors in a number of transformants expressing different combinations of crt genes. Homology suggests that crtE may be involved in binding to the isoprenoid substrates and/or in the common oxidation reactions.	complete CACAO 2968

References

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