Paper chromatography of amino acids and other organic compounds in selected solvents

Edwards, Cecile H.; Gadsden, Evelyn L.; Carter, Lolla P.; Edwards, Gerald A.

doi:10.1016/s0021-9673(01)86279-8

Cited by 13 publications

(8 citation statements)

References 6 publications

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“…Chromatography. Amino acids were separated by paper chromatography by using the solvent systems of Wolfe (36) for two-dimensional chromatograms and the butanol-propionic acid-water system of Edwards et al (17) for separations in one dimension. Radioactive areas on paper chromatograms were located by preparing radioautographs or by using a windowless, gas-flow, strip scanner.…”

Section: Methodsmentioning

confidence: 99%

Biosynthesis of α-Ketoglutarate by the Reductive Carboxylation of Succinate inBacteroides ruminicola

Allison

Robinson

1970

J Bacteriol

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Experiments with growing cells and with cell-free extracts of Bacteroides ruminicola indicate that this anaerobic bacterium can synthesize α-ketoglutarate by a reductive carboxylation of succinate. When the organism was grown in medium containing succinate- 1,4 - 14 C , most of the radioactivity in cells was in the protein fraction and most of the 14 C in protein was in the glutamic acid family of amino acids (glutamate, proline, and arginine). When unlabeled succinate was added to culture medium containing glucose- U - 14 C , incorporation of radioactivity into the glutamic acid family of amino acids was greatly reduced. This supports the concept that succinate is an intermediate in synthesis of α-ketoglutarate. Cell-free extracts of the organism incubated with succinate- 1,4 - 14 C incorporated 14 C into amino acids and most of this was found in glutamate. The cofactors which stimulate glutamate synthesis from succinate by extracts from these cells appear to be similar to the factors that have been demonstrated with extracts from photosynthetic bacteria. The position of label in glutamate synthesized from succinate- 1,4 - 14 C , the probable absence of isocitric dehydrogenase, and studies with labeled citrate and with inhibitors of citric acid cycle enzymes support the concept of a reductive carboxylation of succinate as the only, or at least a major, mechanism for synthesis of α-ketoglutarate in this organism. This appears to be the first evidence for a net synthesis of α-ketoglutarate by this reaction in a nonphotosynthetic heterotrophic organism.

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Section: Methodsmentioning

confidence: 99%

Biosynthesis of α-Ketoglutarate by the Reductive Carboxylation of Succinate inBacteroides ruminicola

Allison

Robinson

1970

J Bacteriol

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“…solvent on Whatman no. 1 paper (Edwards, Gadsden, Carter & Edwards, 1959) revealed the presence of only glutamate and aspartate.…”

Section: Experimental and Resultsmentioning

confidence: 97%

“…The crystalline material was identified as L( + )glutamic acid hydrochloride by paper chromatography (Edwards et al 1959), paper electrophoresis (Frahn & Mills, 1964), melting point (169-171.5o, uncorrected, authentic material m.p. 170-172°, uncorrected), mixed melting point (168-171.5°, un- Table 5.…”

Section: Experimental and Resultsmentioning

confidence: 99%

Metabolism of propionate by sheep liver. Stimulation of the mitochondrial rate by factors from the cell sap

Smith

Osborne-White

Russell

1965

Biochemical Journal

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1. The rate of metabolism of propionate by aged sheep-liver mitochondria in the presence of oxygen + carbon dioxide (95:5) was 5.0 (+/- s.e.m. 0.8) mumoles/mg. of mitochondrial N/hr. 2. When aged in the presence of the mitochondrial supernatant the rate was increased. Mitochondria from 0.33g. of liver, when combined with the corresponding mitochondrial supernatant from 0.08g. of liver, metabolized propionate at a rate of 11.4 (+/- s.e.m. 1.2) mumoles/mg. of mitochondrial N/hr. This rate is comparable with rates previously obtained with aged nuclear-free homogenates. 3. Two factors in the mitochondrial supernatant were detected, which when combined reproduced the effect of the fresh supernatant and prevented loss of activity on aging. One of these was non-diffusible and was recovered by fractionation of the dialysed mitochondrial supernatant with ammonium sulphate. The second factor was present in an ultrafiltrate of fresh mitochondrial supernatant and in boiled mitochondrial supernatant; it was isolated and identified as l(+)-glutamate. 4. The effect of the non-diffusible factor was due to protection of the mitochondria from the aging process, whereas glutamate served both in this capacity and as a direct stimulant of propionate metabolism at low concentration.

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“…Many investigators working with the chromatography of fatsoluble vitamins have impregnated their filter papers with alumina [A1s(S04) ], coated them with vaseline or paraffin, or treated them in other ways (5, 27). There was no attempt to treat the papers specially, as we wanted to secure R¡ values for the vitamins using the same procedures as were applied in the two-dimensional chroma- -.80 tography of other compounds in our experiments (14). Our failure to obtain discrete spots may have been due…”

Section: Resultsmentioning

confidence: 99%

Paper Chromatography of Certain Vitamins in Phenol and Butanol-Propionic Acid-Water Solvents

Gadsden¹,

Edwards²,

Edwards³

1960

Anal. Chem.

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Sitmmia,ry. Phloridzin, the main phenolic glucoside in apple leaves, has I)een founid to uinidergo transform-nation (lulringi chromatograp)hy. WhlleIn chromatographed repeatedfly in ammoniacal solvents, at least 2 new derivatives appeared. One of these vas identified as phloretic acid. When bioassayed in the presence of indole-3-a,cetic acid this substance l)ehaved as though it promoted the destruction of the auxin. Comparative bioassay with naphthaleneacetic acid suggested that phloretic acid acts on indoleacetic acid desAtruction via stimulation of indoleacetic acid oxidase. However, at low concentration and ini presence of a swall am,ount of phloridzin it also showed a synergistic effect with indole-acetic acid. A substance with the same characteristics was obtained directly from apple leaves, which are known to contain phloridzin when the extracts wvere chromatographed only once in the same (alkaline) solvent. While lnot completely confirmed, this suggests that phloretic acid is niormally present in apple leaves, where it may affect growth there by promoting indoleacetic acid oxidation Phlori(lzini, the miiain phenolic glucoside in apple (6. 16, 22), is present as a growth inhilbitor in a relationship with flower bud formation in trees of 2 apple varieties (3, 4). Its appearance in the leaves seemed to be connected with the cessation of long-slhoot growth onI the trees. The strong inhibitorv effect of 1)hloridzin on1 the growth of plant tissues has been explained by Stenli(d as a stimulation of indlole-3-acetic aci(d oxidase action (5, 18). Onl the other hand, phloridzin has been shlowIn by Marre (10, 11) to (lecrease markedly the respiration of Elodea leaves buit to (louble their phloto-synthetic rate. The additional. interesting feature of this glucoside is its lability (12). The instability of phloridzin is so great that it un(lergoes tranisformation even during chromatographv in alkaline solvent. Solvents con-taiiming amm-nonia have been frequently used to develop chromiiatograms of natural auxins and inhibitors because of 'better separation of these substances on the lpal)er (1. 9, 19). It is, however, a (lisa(lvantage that the hilgh pH favors the traansforimlatioin of somiie of the chrornatographed compileounids. Indole-pyruivic .ci(l uIIdergoes coml)lex chanlges in alkaliine solvenlts (7), anl(indoleacetyl-glucosidle is comnlpletely lby-(drolvze(d (23).

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Paper chromatography of amino acids and other organic compounds in selected solvents

Cited by 13 publications

References 6 publications

Biosynthesis of α-Ketoglutarate by the Reductive Carboxylation of Succinate inBacteroides ruminicola

Biosynthesis of α-Ketoglutarate by the Reductive Carboxylation of Succinate inBacteroides ruminicola

Metabolism of propionate by sheep liver. Stimulation of the mitochondrial rate by factors from the cell sap

Paper Chromatography of Certain Vitamins in Phenol and Butanol-Propionic Acid-Water Solvents

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