Carbon dioxide fixation in green sulphur bacteria
TL;DR: It is concluded that in washed suspensions, CO(2) is fixed mainly by a mechanism involving the reductive carboxylic acid cycle, and appears to exhibit a similar pattern to that in C. thiosulfatophilum strain 8346.
Abstract: 1. About one-third of the CO2 fixed during photosynthesis by washed suspensions of Chlorobium thiosulfatophilum strain 8346 gave rise to α-oxoglutarate and branched-chain oxo acids, mainly β-methyl-α-oxovalerate. Another one-third to one-half gave rise to a polyglucose. 2. The fixation of CO2 was inhibited by fluoroacetate, increasing concentrations up to 1mm stimulating the accumulation of α-oxoglutarate and causing a decrease in the formation of the branched-chain oxo acids and polyglucose. 3. Acetate was converted into the same products as was CO2. 4. Fluoroacetate (1mm) had a negligible effect on the formation of polyglucose from acetate and caused a slight inhibition of the formation of the branched-chain oxo acids and increased accumulation of α-oxoglutarate. 5. Iodoacetate (1mm) strongly inhibited polyglucose formation from acetate and caused accumulation of pyruvate. The formation of the branched-chain oxo acids from acetate was only slightly affected by this inhibitor. 6. Pyruvate can be metabolized by this organism in the presence of a suitable electron donor whether CO2 is present or not. In the absence of CO2 pyruvate is converted into polyglucose. 7. The accumulation of oxo acids during CO2 fixation is completely inhibited by NH4+ ions. The formation of the branched-chain oxo acids is considerably decreased by the presence of isoleucine, leucine or valine, or a mixture of these. 8. CO2 fixation in two other strains of Chlorobium appears to exhibit a similar pattern to that in C. thiosulfatophilum strain 8346. 9. It is concluded that in washed suspensions, CO2 is fixed mainly by a mechanism involving the reductive carboxylic acid cycle. Acetate, the product of the cycle, is converted into polyglucose via pyruvate synthase and a reversal of glycolysis or into branched-chain oxo acids by an unknown mechanism.
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TL;DR: This chapter explains the description and physiology of one-carbon-utilizing micro-organisms along with their energy metabolism and carbon assimilation, and considers mechanisms that offer a solution to the problem of net biosynthesis from one- carbon units at reduction levels ranging from carbon dioxide to methane.
Abstract: Publisher Summary One-carbon compounds occur abundantly at all oxidation levels between methane and carbon dioxide. Methane occurs in coal and oil deposits and is also evolved on a large scale as an end-product of many fermentations. Carbon dioxide is also abundantly present in the atmosphere, in natural waters, and as carbonates in the earth. A considerable number of micro-organisms have developed the ability to utilize such compounds as carbon or energy sources. This chapter examines these micro-organisms and the biochemical problems, which are posed by energy transduction and biosynthesis of cell constituents from the one-carbon substrate. It explains the description and physiology of one-carbon-utilizing micro-organisms along with their energy metabolism and carbon assimilation. The utilization of one-carbon compounds is largely confined to prokaryotic organisms. Many enzymes that catalyze the oxidation of one-carbon compounds at a variety of oxidation levels are also extensively studied. The chapter also considers mechanisms that offer a solution to the problem of net biosynthesis from one-carbon units at reduction levels ranging from carbon dioxide to methane.
211 citations
TL;DR: The findings that led the group to the discovery of the reductive carboxylic acid cycle, the nature and resolution of the controversy that followed, and the possible evolutionary implications of the cycle as an ancient mechanism for photosynthetic CO2 assimilation that preceded the pentose cycle and served as a precursor of the Krebs cycle in aerobic metabolism are discussed.
Abstract: The Krebs cycle (citric acid or tricarboxylic acid cycle), the final common pathway in aerobic metabolism for the oxidation of carbohydrates, fatty acids and amino acids, is known to be irreversible. It liberates CO2 and generates NADH whose aerobic oxidation yields ATP but it does not operate in reverse as a biosynthetic pathway for CO2 assimilation. In 1966, our laboratory described a cyclic pathway for CO2 assimilation (Evans, Buchanan and Arnon 1966) that was unusual in two respects: (i) it provided the first instance of an obligate photoautotroph that assimilated CO2 by a pathway different from Calvin's reductive pentose phosphate cycle (Calvin 1962) and (ii) in its overall effect the new cycle was a reversal of the Krebs cycle. Named the 'reductive carboxylic acid cycle' (sometimes also called the reductive tricarboxylic acid cycle) the new cycle appeared to be the sole CO2 assimilation pathway in Chlorobium thiosulfatophilum (Evans et al. 1966) (now known as Chlorobium limicola forma thiosulfatophilum). Chlorobium is a photosynthetic green sulfur bacterium that grows anaerobically in an inorganic medium with sulfide and thiosulfate as electron donors and CO2 as an obligatory carbon source. In the ensuing years, the new cycle was viewed with skepticism. Not only was it in conflict with the prevailing doctrine that the 'one important property ... shared by all (our emphasis) autotrophic species is the assimilation of CO2 via the Calvin cycle' (McFadden 1973) but also some of its experimental underpinnings were challenged. It is only now that in the words of one of its early skeptics (Tabita 1988) 'a long and tortuous controversy' has ended with general acceptance of the reductive carboxylic acid cycle as a photosynthetic CO2 assimilation pathway distinct from the pentose cycle. (Henceforth, to minimize repetitiveness, the reductive pentose phosphate cycle will often be referred to as the pentose cycle and the reductive carboxylic acid cycle as the carboxylic acid cycle.) Aside from photosynthetic pathways which are the focus of this article, CO2 assimilation is also known to sustain autotrophic growth via the acetyl-CoA pathway (Wood et al. 1986). Our aim here is to discuss (i) the findings that led our group to the discovery of the reductive carboxylic acid cycle, (ii) the nature and resolution of the controversy that followed, and (iii) the possible evolutionary implications of the cycle as an ancient mechanism for photosynthetic CO2 assimilation that preceded the pentose cycle and served as a precursor of the Krebs cycle in aerobic metabolism.
192 citations
TL;DR: The results suggest that Chromatium and R. rubrum assimilate CO2 mainly via ribulose 1,5-diphosphate carboxylase and the associated reactions of the reductive pentose phosphate cycle, whereas Chlorobium utilizes other mechanisms.
Abstract: 1. The carbon isotope discrimination properties of a representative of each of the three types of photosynthetic bacteria Chlorobium thiosulfatophilum, Rhodospirillum rubrum and Chromatium and of the C3-alga Chlamydomonas reinhardii were determined by measuring the ratio of 13CO2 to 12CO2 incorporated during photoautotrophic growth. 2. Chromatium and R. rubrum had isotope selection properties similar to those of C3-plants, whereas Chlorobium was significantly different. 3. The results suggest that Chromatium and R. rubrum assimilate CO2 mainly via ribulose 1,5-diphosphate carboxylase and the associated reactions of the reductive pentose phosphate cycle, whereas Chlorobium utilizes other mechanisms. Such mechanisms would include the ferredoxin-linked carboxylation enzymes and associated reactions of the reductive carboxylic acid cycle.
187 citations
TL;DR: The physiological aspects regulating hydrogen metabolism in anoxygenic phototrophic bacteria, along with technical and economical aspects of hydrogen production, with particular reference to purple nonsulfur bacteria are explained.
Abstract: Publisher Summary The chapter focuses on the physiological aspects regulating hydrogen metabolism in anoxygenic phototrophic bacteria, along with technical and economical aspects of hydrogen production, with particular reference to purple nonsulfur bacteria. Studies on the growth phase of the organism are considered equally important as studies on hydrogen production by resting cells; hence, the chapter explains in general the physiology related to hydrogen metabolism and other important related areas of research on anoxygenic phototrophic bacteria. Biological hydrogen evolution is still in an exploratory stage in bioenergy production research, unlike methane (biogas) and alcohol production. There is enough justification to carry out further work that now concentrates on process development rather than on strain selection. For this futuristic research, a multi- and interdisciplinary approach is the need of the day.
140 citations
TL;DR: The distribution of the two pathways among the bacteria tested was in general agreement with their previously established phylogenetic relationships and clearly indicates that the five-carbon pathway is the more ancient process, whereas the pathway utilizing ALA synthase probably evolved much later.
Abstract: Two biosynthetic pathways are known for the universal tetrapyrrole precursor, δ-aminolevulinic acid (ALA). In the ALA synthase pathway which was first described in animal and some bacterial cells, the pyridoxal phosphate-dependent enzyme ALA synthase catalyzes condensation of glycine and succinyl-CoA to form ALA with the loss of C-1 of glycine as CO2. In the five-carbon pathway which was first described in plant and algal cells, the carbon skeleton of glutamate is converted intact to ALA in a proposed reaction sequence that requires three enzymes, tRNAGlu, ATP, Mg2+, NADPH, and pyridoxal phosphate. We have examined the distribution of the two ALA biosynthetic pathways among various genera, using cell-free extracts obtained from representative organisms. Evidence for the operation of the five-carbon pathway was obtained by the measurement of RNase-sensitive label incorporation from glutamate into ALA, using 3,4-[3H]glutamate or 1-[14C]glutamate as substrate. ALA synthase activity was indicated by RNase-insensitive incorporation of label from 2-[14C]glycine into ALA. The distribution of the two pathways among the bacteria tested was in general agreement with their previously established phylogenetic relationships and clearly indicates that the five-carbon pathway is the more ancient process, whereas the pathway utilizing ALA synthase probably evolved much later. The five-carbon pathway is apparently the more widely utilized one among bacteria, while the ALA synthase pathway seems to be limited to the α subgroup of purple bacteria.
135 citations