• 4,4′ diapolycopene-dioate ester;
  • antioxidative properties;
  • C30 carotenoid biosynthesis pathway;
  • red-pigmented Bacillus firmus isolates



Pigmented Bacillus spp. with probiotic properties have been isolated. In the yellow-/orange-coloured strains, the carotenoid pigments present have been characterized. In contrast, the carotenoids present in the Bacillus isolates coloured red await identification. The present article reports progress on the elucidation of the pigment biosynthetic pathway in these red-pigmented Bacillus firmus strains.

Methods and Results

A combination of UV/Vis, chromatographic and mass spectrometry (MS) has revealed the properties of the predominant pigment and the end-point carotenoid of the pathway to be methyl 4,4′-diapolycopene-dioate after transmethylation. The diglycosyl ester of 4,4′-diapolycopene-dioate persists in vivo prior to chemical treatment. Different mutants and inhibitor treatment were employed to establish the C30 biosynthesis pathway with all precursors and intermediates to 4,4′-diapolycopene-dioate detected, which include 4,4′-diapophytene and all desaturation intermediates to 4,4′-diapolycopene and 4,4′-diapolycopene-dialdehyde. To cultures synthesizing the 4,4′-diapolycopene-dioate derivative and those in which its formation was inhibited, oxidative stress was induced by peroxide treatment. Conditions that decreased the growth rate of the pigmented cells by only 30% caused a complete growth inhibition of the culture devoid of the 4,4′-diapolycopene-dioate derivative.


This finding demonstrates the diversity of C30 carotenoid biosynthesis in Bacillus species and the antioxidative function of the 4,4′-diapolycopene-dioate derivative in B. firmus cells.

Significance and Impact of the Study

It could be shown that the C30 4,4′-diapolycopene-dioate derivatives protect pigmented B. firmus from peroxidative reactions. Under oxidative conditions, this can be an ecological advantage over nonpigmented (=noncarotenogenic) strains that are equally abundant.