THE METABOLISM OF ACETATE BY WHEAT STEM RUST UREDOSPORES

1964 ◽  
Vol 42 (6) ◽  
pp. 883-888 ◽  
Author(s):  
S. Suryanarayanan ◽  
W. B. McConnell

When uredospores of Puccinia graminis var. tritici (race 15B) were incubated at pH 6.2 in phosphate buffer containing either acetate-1-C14or -2-C14, about 12% of the radioactivity was removed from the solution in a period of 3 hours. Respired carbon dioxide contained about 45% and 22% of the carbon-14 taken up as acetate-1-C14and acetate-2-C14, respectively. Incorporation of carbon-14 into spore components was considerably higher with acetate-2-C14than with acetate-1-C14. With either tracer most of the radioactivity in water-soluble spore materials was accounted for in amino acids and neutral substances. Glutamic acid was particularly radioactive and accounted for about 40% of the radioactivity in the amino acid fraction. Incorporation of carbon-14 into the glutamic acid skeleton was consistent with the view that both the tricarboxylic acid cycle and the glyoxalate cycle were functioning.

1961 ◽  
Vol 39 (10) ◽  
pp. 1559-1566 ◽  
Author(s):  
H. Reisener ◽  
W. B. McConnell ◽  
G. A. Ledingham

When uredospores of Puccinia graminis var. tritici race 15B were shaken in a medium containing M/30 phosphate buffer, pH 6.2, and valerate-1-C14, 97% of the radioactivity was removed from the solution in a period of 3 hours. Fifty-five per cent of the carbon-14 was released as carbon dioxide, and 42% was incorporated into the spores. Carbon-14 was found in many cellular components but the water-soluble fraction accounted for 48% of the tracer in the spores. About two thirds of the water-soluble carbon-14 was in a fraction containing amino acids, amides, and peptides, with glutamic acid, glutamine, and γ-aminobutyric acid being highly radioactive. Carbon-5 of glutamic acid and carbon-1 of γ-aminobutyric acid were particularly radioactive. In addition carbon-1 of glutamic acid was appreciably radioactive. The results are consistent with the view that γ-aminobutyric acid was formed by decarboxylation of glutamic acid and that glutamic acid became labelled as a result of β-oxidation of the valerate-1-C14 to yield acetate-1-C14 which in turn was metabolized by the tricarboxylic acid cycle.


1963 ◽  
Vol 41 (1) ◽  
pp. 1-7 ◽  
Author(s):  
H. Reisener ◽  
A. J. Finlayson ◽  
W. B. McConnell

When uredospores of Puccinia graminis var. tritici race 15B were shaken in a medium containing M/30 phosphate buffer, pH 6.2, and valerate-2-C14, about 88% of the radioactivity was removed from the buffer solution in a period of 3 hours. About 40% of the carbon-14 taken from the buffer was found in a water-soluble extract of the spores and about 15% was respired as carbon dioxide. The result is compared with an earlier report that carbon 1 of valerate is more extensively released as carbon dioxide and less extensively incorporated into spore components. Glutamic acid, glutamine, γ-aminobutyric acid, and alanine of high specific activity were isolated. It was estimated from partial degradation that more than one-half of the carbon-14 of glutamic acid occurred in position 4 and that carbon 5 was very weakly labelled. Citric acid was also of high specific activity and was labelled predominantly in the internal carbons.It is concluded that respiring rust spores utilize externally supplied valerate by β-oxidation, which releases carbons 1 and 2 in a form which is metabolized as acetate by the tricarboxylic acid cycle.


1963 ◽  
Vol 41 (1) ◽  
pp. 1-7 ◽  
Author(s):  
H. Reisener ◽  
A. J. Finlayson ◽  
W. B. McConnell

When uredospores of Puccinia graminis var. tritici race 15B were shaken in a medium containing M/30 phosphate buffer, pH 6.2, and valerate-2-C14, about 88% of the radioactivity was removed from the buffer solution in a period of 3 hours. About 40% of the carbon-14 taken from the buffer was found in a water-soluble extract of the spores and about 15% was respired as carbon dioxide. The result is compared with an earlier report that carbon 1 of valerate is more extensively released as carbon dioxide and less extensively incorporated into spore components. Glutamic acid, glutamine, γ-aminobutyric acid, and alanine of high specific activity were isolated. It was estimated from partial degradation that more than one-half of the carbon-14 of glutamic acid occurred in position 4 and that carbon 5 was very weakly labelled. Citric acid was also of high specific activity and was labelled predominantly in the internal carbons.It is concluded that respiring rust spores utilize externally supplied valerate by β-oxidation, which releases carbons 1 and 2 in a form which is metabolized as acetate by the tricarboxylic acid cycle.


1957 ◽  
Vol 35 (1) ◽  
pp. 1259-1266 ◽  
Author(s):  
W. B. McConnell

Radioactive wheat seeds, obtained by injecting acetate-C14 into the stems of the parent plants, were germinated in the absence of light and nutrient and the fate of the carbon-14 was observed. Carbon respired as carbon dioxide had a higher specific activity than any of the major seed components except protein. Variations were found in the patterns by which material was transferred from the kernel to new tissue as reflected in a comparison of the activity of various components. Glutamic acid was the most active compound isolated either from the original seeds or from the new tissues. This observation, together with similarities noted in the intramolecular distribution of carbon-14 in glutamic acid of new tissue and seed residues, indicated that glutamic acid was reutilized for the biosynthesis of seedling protein. Changes in the labelling of glutamic acid during transfer to new tissue are qualitatively in accord with the idea that at least some of the amino acid is used after re-entry into the tricarboxylic acid cycle.


1957 ◽  
Vol 35 (12) ◽  
pp. 1259-1266 ◽  
Author(s):  
W. B. McConnell

Radioactive wheat seeds, obtained by injecting acetate-C14 into the stems of the parent plants, were germinated in the absence of light and nutrient and the fate of the carbon-14 was observed. Carbon respired as carbon dioxide had a higher specific activity than any of the major seed components except protein. Variations were found in the patterns by which material was transferred from the kernel to new tissue as reflected in a comparison of the activity of various components. Glutamic acid was the most active compound isolated either from the original seeds or from the new tissues. This observation, together with similarities noted in the intramolecular distribution of carbon-14 in glutamic acid of new tissue and seed residues, indicated that glutamic acid was reutilized for the biosynthesis of seedling protein. Changes in the labelling of glutamic acid during transfer to new tissue are qualitatively in accord with the idea that at least some of the amino acid is used after re-entry into the tricarboxylic acid cycle.


1963 ◽  
Vol 41 (3) ◽  
pp. 737-743 ◽  
Author(s):  
H. Reisener ◽  
A. J. Finlayson ◽  
W. B. McConnell ◽  
G. A. Ledingham

When uredospores of wheat stem rust were shaken for 3 hours with phosphate buffer (pH 6.2) containing propionate-1-C14, -2-C14, or -3-C14, about 55% of the carbon-14 was removed from the solution. With propionate-1-C14, most of the carbon-14 taken up was released as carbon dioxide-C14, whereas about 20% and 31% of propionate carbon 2 and carbon 3, respectively, was incorporated into the spores. The specific activity of a fraction consisting of the free amino acids of a hot-alcohol and hot-water extract of the spores increased markedly with increase in the position number of propionate in which the carbon-14 was located. A similar relation was observed for other fractions such as soluble carbohydrates, ether-soluble material, organic acids, and insoluble residue from spores. The most active amino acids isolated were glutamic acid, γ-aminobutyric acid, and alanine. Partial degradations showed that with propionate-2-C14 the carboxyl groups of glutamic acid were especially radioactive, whereas with propionate-3-C14 the internal carbons were most radioactive.It is concluded that propionate metabolism in the rust spores involved conversion of carbon 1 to carbon dioxide, and utilization of carbons 2 and 3 as acetate with carbon 2 behaving as the carboxyl carbon.


2021 ◽  
Author(s):  
E-Bin Gao ◽  
Penglin Ye ◽  
Haiyan Qiu ◽  
Junhua Wu ◽  
Huayou Chen

Abstract Background: The outstanding ability of directly assimilating carbon dioxide and sunlight to produce biofuels and chemicals impels photosynthetic cyanobacteria to become attractive organisms for the solution to the global warming crises and the world energy growth. The cyanobacteria-based method for ethanol production has been increasingly regarded as alternatives to food biomass-based fermentation and traditional petroleum-based production. Therefore, we engineered the model cyanobacterium Synechocystis sp. PCC 6803 to synthesize ethanol and optimized the biosynthetic pathways for improving ethanol production under photoautotrophic conditions.Results: In this study, we successfully achieved the photosynthetic production of ethanol from atmospheric carbon dioxide by an engineered mutant Synechocystis sp. PCC 6803 with over-expressing the heterologous genes encoding Zymomonas mobilis pyruvate decarboxylase (PDC) and Escherichia coli NADPH-dependent alcohol dehydrogenase (YqhD). The engineered strain was further optimized by an alternative engineering approach to improve cell growth, and increase the intracellular supply of the precursor pyruvate for ethanol production under photoautotrophic conditions. This approach includes blocking phosphoenolpyruvate synthetic pathway from pyruvate, removing glycogen storage, and shunting carbon metabolic flux of tricarboxylic acid cycle. Through redirecting and optimizing the metabolic carbon flux of Synechocystis, a high ethanol-producing efficiency was achieved (248 mg L-1 day-1) under photoautotrophic conditions with atmospheric CO2 as the sole carbon source. Conclusions: The engineered strain SYN009 (∆slr0301/pdc-yqhD, ∆slr1176/maeB) would become a valuable biosystem for photosynthetic production of ethanol and for expanding our knowledge of exploiting cyanobacteria to produce value chemicals directly from atmospheric CO2.


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