| Summary: | Carotenoids are extensively used in drugs, cosmetics, nutrients, and foods, owing to their antioxidant and anti-inflammatory characteristics. Diverse marine heterotrophic prokaryotes can accumulate carotenoids and become promising alternatives for the advancement of carotenoids production. In this research, 55 strains were isolated and cultivated from tidal flat sediment in Zhoushan and classified into the phyla <i>Pseudomonadota</i> (<i>n</i> = 24), <i>Bacillota</i> (<i>n</i> = 18), <i>Bacteroidota</i> (<i>n</i> = 9), and <i>Actinomycetota</i> (<i>n</i> = 4). Nine of them accumulated carotenoids, and most of them belonged to the families <i>Flavobacteriaceae</i> (<i>n</i> = 4) and <i>Erythrobacteraceae</i> (<i>n</i> = 4). Among those carotenoid-producing strains, one strain, designated as D39<sup>T</sup>, was proposed as one novel species belonging to the genus <i>Croceibacterium</i> through polyphasic taxonomy approaches. Genomic annotations and carotenoid compound determinations revealed that strain D39<sup>T</sup> encoded <i>crtEBIYZG</i> genes and mainly accumulated zeaxanthin as major carotenoids. Furthermore, carotenoid biosynthesis pathway in the majority of <i>Croceibacterium</i> strains were identical with that in the strain D39<sup>T</sup>, implying that <i>Croceibacterium</i> members can be sources of producing zeaxanthin. This study enhances knowledge of microbial biodiversity in tidal flats, proposes a novel carotenoid-producing <i>Croceibacterium</i> species, and elucidates carotenoid biosynthesis pathway in the genus <i>Croceibacterium</i>, which contribute to enriching marine carotenoid-producing strains and promoting a comprehensive insight into genomic contents of them.
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