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Most  Sphingomonas  species synthesize the yellow carotenoid nostoxanthin. However, the carotenoid biosynthetic pathway of these species remains unclear. In this study, we cloned and characterized a carotenoid biosynthesis gene cluster containing four carotenogenic genes ( crtG ,  crtY ,  crtI  and  crtB ) and a β-carotene hydroxylase gene ( crtZ ) located outside the cluster, from the gellan-gum producing bacterium  Sphingomonas elodea  ATCC 31461. Each of these genes was inactivated, and the biochemical function of each gene was confirmed based on chromatographic and spectroscopic analysis of the intermediates accumulated in the knockout mutants. Moreover, the  crtG  gene encoding the 2,2′-β-hydroxylase and the  crtZ  gene encoding the β-carotene hydroxylase, both responsible for hydroxylation of β-carotene, were confirmed by complementation studies using  Escherichia coli  producing different carotenoids. Expression of  crtG  in zeaxanthin and β-carotene accumulating  E. coli  cells resulted in the formation of nostoxanthin and 2,2′-dihydroxy-β-carotene, respectively. Based on these results, a biochemical pathway for synthesis of nostoxanthin in  S. elodea  ATCC 31461 is proposed.

Cloning and Characterization of Genes Involved in Nostoxanthin Biosynthesis of Sphingomonas elodea ATCC 31461