Formation of maxima in the absorption spectrum of carotenoids in the region around 370 nm; consequences for the interpretation of certain action spectra

Abstract

Most carotenoids show a 3-peak-absorption curve in the visible spectral region in polar solutions. The addition of a definite quantity of H₂O to such solutions (ethanol, methanol, aceton, isopropanol) changes the absorption curve of these pigments in a characteristic manner. A new peak appears in the uv region of the spectrum (e.g.in the case of lutein at 370 nm); simultaneously the 3-peak fine structure of the visible spectrum diminishes and completely disappears after further addition of H₂O. Such changes are observed especially in the case of lutein and zeaxanthin, but also in the case of neoxanthin, violaxanthin and lycopene (of the carotenoids analyzed). During thermic excitation (45° C) the uv-peak in the carotenoid spectrum disappears and the normal 3-peak curve is restored; upon cooling the uv-peak appears again. The variation of the carotenoid spectrum and the formation of a maximum in the uv-region are possibly caused by an aggregation of the pigment molecules with participation of H₂O molecules. This formation of polymers obviously leads to an alteration in the distribution of electrons in the chromophore system of the carotenoid molecule and thereby to a change of the light absorption. Water-soluble carotenoid complexes isolated from spinach chloroplasts show a strong light absorption in the uv-region and a one-peak absorption curve in the visible blue. After transfer of the complex to polar solutions a characteristic 3-peak carotenoid curve appears in the blue region of the spectrum; concomitantly the maximum in the uv disappears. That means that carotenoids which are bound to membranes or particles in the intact cell may have a 4-peak absorption curve similar to that of pigments which are dissolved in the water-containing alcohols mentioned above. It is conceivable that those carotenoids which do not form uv peaks in the dissolved state are able to do so under conditions under which carotenoids are bound to membranes or particles. The similarity of some action spectra to certain 4-peak, carotenoid spectra is striking. This is true particularly for the action spectrum of the first positive curvature of Avena coleoptile (Fig. 10). On the basis of the described abilities of the carotenoids to form an absorption peak in the wave uv, the appearance of such a maximum in an action spectrum (in the region about 370 nm) can no longer be considered to be sufficient proof for the participation of a flavin as light-acceptor.