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Article Dans Une Revue Microbial Cell Factories Année : 2022

Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa

Anesu Conrad Moyo
Daniele Giuffrida
  • Fonction : Auteur
Leonardo Joaquim van Zyl
  • Fonction : Auteur
Marla Trindade

Résumé

Background: The genus Planococcus is comprised of halophilic bacteria generally reported for the production of carotenoid pigments and biosurfactants. In previous work, we showed that the culturing of the orange-pigmented Planococcus sp. CP5-4 isolate increased the evaporation rate of industrial wastewater brine effluent, which we attributed to the orange pigment. This demonstrated the potential application of this bacterium for industrial brine effluent management in evaporation ponds for inland desalination plants. Here we identified a C 30-carotenoid biosynthetic gene cluster responsible for pigment biosynthesis in Planococcus sp. CP5-4 through isolation of mutants and genome sequencing. We further compare the core genes of the carotenoid biosynthetic gene clusters identified from different Planococcus species' genomes which grouped into gene cluster families containing BGCs linked to different carotenoid product chemotypes. Lastly, LC-MS analysis of saponified and unsaponified pigment extracts obtained from cultures of Planococcus sp. CP5-4, revealed the structure of the main (predominant) glucosylated C 30-carotenoid fatty acid ester produced by Planococcus sp. CP5-4. Results: Genome sequence comparisons of isolated mutant strains of Planococcus sp. CP5-4 showed deletions of 146 Kb and 3 Kb for the non-pigmented and "yellow" mutants respectively. Eight candidate genes, likely responsible for C 30-carotenoid biosynthesis, were identified on the wild-type genome region corresponding to the deleted segment in the non-pigmented mutant. Six of the eight candidate genes formed a biosynthetic gene cluster. A truncation of crtP was responsible for the "yellow" mutant phenotype. Genome annotation revealed that the genes encoded 4,4′-diapolycopene oxygenase (CrtNb), 4,4′-diapolycopen-4-al dehydrogenase (CrtNc), 4,4′-diapophytoene desaturase (CrtN), 4,4′-diaponeurosporene oxygenase (CrtP), glycerol acyltransferase (Agpat), family 2 glucosyl transferase 2 (Gtf2), phytoene/squalene synthase (CrtM), and cytochrome P450 hydroxylase enzymes. Carotenoid analysis showed that a glucosylated C 30-carotenoid fatty acid ester, methyl 5-(6-C 17:3)-glucosyl-5, 6′-dihydro-apo-4, 4′-lycopenoate was the main carotenoid compound produced by Planococcus sp. CP5-4. Conclusion: We identified and characterized the carotenoid biosynthetic gene cluster and the C 30-carotenoid compound produced by Planococcus sp. CP5-4. Mass-spectrometry guided analysis of the saponified and unsaponified pigment extracts showed that methyl 5-glucosyl-5, 6-dihydro-apo-4, 4′-lycopenoate esterified to heptadecatrienoic acid (C 17:3). Furthermore, through phylogenetic analysis of the core carotenoid BGCs of Planococcus species we show
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hal-04148733 , version 1 (03-07-2023)

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Anesu Conrad Moyo, Laurent Dufossé, Daniele Giuffrida, Leonardo Joaquim van Zyl, Marla Trindade. Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa. Microbial Cell Factories, 2022, 21, ⟨10.1186/s12934-022-01752-1⟩. ⟨hal-04148733⟩
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