The accumulation of bacteriochlorophyll precursors by mutant and wild-type strains of Rhodopseudomonas spheroides

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Abstract
1. Two mutant strains of Rhodopseudomonas spheroides, which are blocked in the synthesis of bacteriochlorophyll, accumulate pigments. These have been tentatively identified as magnesium 2,4-divinylphaeoporphyrin a(5) monomethyl ester and the magnesium derivative of 2-devinyl-2-hydroxyethyl-phaeophorbid a, formed by mutant 2/73 and 2/21 respectively. 2. Maximum extracellular production of these pigments occurs when suspensions of the organisms are incubated with low aeration in a growth medium containing iron and supplemented with glycine, succinate, methionine and Tween 80. 3. Concomitant protein synthesis is required for pigment production by the mutants from glycine and succinate but this requirement is less marked when delta-aminolaevulic acid is the substrate. 4. In the absence of Tween 80, a considerable proportion of the total pigment is retained within the cells and appears in the particulate fraction of cell-free extracts. 5. Suspensions of the parent strain containing delta-aminolaevulic acid can be made to accumulate extracellular pigments which are tentatively identified as magnesium protoporphyrin monomethyl ester and the magnesium derivative of 2-devinyl-2-hydroxyethyl-phaeophorbid a. 6. Maximum production occurs with cells incubated photosynthetically after a period of oxygen repression of bacteriochlorophyll synthesis. Formation of the phaeophorbid derivative is enhanced by 8-azaguanine or 5-fluorouracil, or by adenine deficiency in a nutritional mutant; Tween 80 is also needed and iron is essential. 7. Synthesis of bacteriochlorophyll might possibly involve the participation of lipoprotein-bound intermediates, which may be formed at the initial stage of condensation between glycine and succinyl-CoA to give delta-aminolaevulic acid.