Abstract
WHEN cultures of Chlorella vulgaris were incubated with formate-14C it could be demonstrated that the carbon atom of formate may serve as a precursor of the methyl ester carbon atom of chlorophyll a, without contributing extensively to the biosynthesis of the dihydroporphyrin ring. The procedure for growing Chlorella vulgaris as well as the preparation of methyl-phæophorbide a has been described1. However, in the place of inert acetate and glycine added previously, inert sodium formate (2.44 × 10−3 M) as well as 8.6 mgm. (33 μc.) of sodium formate-14C (2.1 × 107 c./m.) were added to the growing medium. Methyl phæophorbide a was converted to chlorin e6 according to the method of Willstätter2, and both degradation products of chlorophyll a were subjected to paper chromatography. Methyl phæophorbide a was chromatographed twice in decane/4-methyl-2-pentanone, 2 : 1 (v/v) and once in n-decane/1-propanol, 6 : 1 (v/v). Repeated chromatography did not significantly alter the specific activity. Chlorin e 6 was chromatographed once in the lutidine/water/ammonia system of Falk et al. 3. The separated pigments were cut out and eluted with acetone and acidified acetone, respectively. Details of this chromatographic procedure will be published elsewhere. The concentration of methyl phæophorbide a or chlorin e 6 in the respective eluate was determined spectrophotometrically, applying the absorption coefficients reported by Holt and Jacobs4 and Stern and Wenderlein5. The solvents of the eluates were evaporated and the solid material deposited as an infinitely thin layer on glass planchettes. The carbon-14 activity was then determined with a windowless methane gas-flow counter operated in the proportional region. The experimental results are shown in Table 1.
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References
Della Rosa, R. J., Altman, K. I., and Salomon, K., J. Biol. Chem., 202, 771 (1953).
Willstätter, R., and Stoll, A., “Investigations on Chlorophyll”, 263 (Science Press Printing Co., Lancaster, Pennsylvania, 1928).
Falk, J., Dresel, E. I. B., Benson, A., and Knight, B. C., Biochem. J., 63, 87 (1956).
Holt, A. S., and Jacobs, E. E., Amer. J. Bot., 41, 710 (1954).
Stern, A., and Wenderlein, H., Z. Phys. Chem., 174, 81 (1935).
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GREEN, M., ALTMAN, K., RICHMOND, J. et al. Role of Formate in the Biosynthesis of Chlorophyll a . Nature 179, 375 (1957). https://doi.org/10.1038/179375a0
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DOI: https://doi.org/10.1038/179375a0
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