Estimation of <scp>l</scp>-malate and fumarate by malic decarboxylase of <i>Lactobacillus arabinosus</i>

Nossal, Peter M

doi:10.1042/bj0500349

Cited by 48 publications

(9 citation statements)

References 11 publications

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“…The preceding results demonstrate that the bacterial method of Nossal (19) and the enzymic method of Hatch (10) Figure 1. [14C1/14C4]malate (35%/65%) was isolated from the products of dark 14CO2 fixation by Kalanchoe plantlets and its label distribution characterized by the bacterial method.…”

Section: Methodsmentioning

confidence: 83%

Malate Synthesis by Dark Carbon Dioxide Fixation in Leaves

Levi¹,

Perchorowicz²,

Gibbs³

1978

Plant Physiol.

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Te rates of dark CO2 fixation and the label distribution in malate following dark '4CO2 fixation in a C4 plant (maize), a C-3 plant (sunflower), and two Crassulacean acid metabolsm plants (BryopbyHum calycinum and KaIlancho diagrematianum leaves and plantiets) are compared. Within the first 30 minutes of dark 1"CO2 fixaton, leaves of maize, B. calycinum, and sunflower, and K. diagremondanum plantlets fix CO2 at rates of 1.4, 3.4, 0.23, and 1.0 Fmoles of C02/mg of chlkopbyfll hour, respectively. Net C02 fixathi stops within 3 hours in maize and sunflower, but Crassulaceans continue fixing CO2 for the duration of the 23-hour experiment.A bacterial procedure using Lactobacilus plantaum ATCC No. 8014 and one using malc enzyme to remove the P-carboxyl The characteristics of photosynthetic C02 fixation in C-3 and C4 plants have been extensively compared with Crassulacean acid metabolism (CAM) in recent years. In contrast, the relationship between dark C02 fixation in C-3 and C4 plants and the dark fixation in CAM plants has received little attention. In general, most investigations on dark C02 fixation have centered about CAM plants. In the characterization of CAM metabolism, rates of C02 uptake (3,4,22) and the label distribution in malate (5-7) have received much emphasis. Degradation of malate by Lctobacillusplantarum was first used by Utter (25) The purpose of this paper is to compare dark C02 fixation in a C-3 plant (sunflower), a C4 plant (maize), and two CAM plants (Kalanchoe diagremontianum and Bryophyllum calycinum) with respect to the rates of dark C02 fixation over a 23-hr period and the distribution of isotope in malate made during this period. In addition, dark C02 fixation in mature and immature (taken from plantlets) CAM leaves is compared. A preliminary report of this research has been published (17). MATERIALS AND METHODSPlant Material. K. diagremontianum and maize (Zea mays, var.Early Fortune) were grown in growth chambers with a regime of 12 hr light at 25 to 27 C followed by 12 hr dark at 18 to 20 C, under cool white fluorescent lamps supplemented with tungsten lamps. B. calycinum and sunflower (Helianthus annuus) were grown in the greenhouse under natural lighting. All plants were maintained on Hoagland solution (1 1).Dark "4CO2 Fixation. Dark C02 fixation was under green safelights. For 14CO2 fixation periods of less than 1 hr, sunflower leaves, Bryophyllum or Kalanchoe leaf discs (1-to 2-cm diameter), the youngest expanded leaf of maize seedlings or derooted Kalanchoe plantlets were tied together with sewing thread and floated on 10 ml of distilled H20 in 50-ml sealed flasks shaken in a water bath maintained at 25 C. Longer exposures (3 hr or more) to 14CO2 were performed in a darkroom at room temperature. '4C02 (5 ml, 2 IAmol/ml, 10 tCi/4tmol) was added by syringe through a serum cap which sealed the flask. The initial concentration of C02 in each flask was about 0.4%. "CO2 fixation was terminated by lifting the leaves by their threads and dropping them into boiling 80%o (v/v)

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Section: Methodsmentioning

confidence: 83%

Malate Synthesis by Dark Carbon Dioxide Fixation in Leaves

Levi¹,

Perchorowicz²,

Gibbs³

1978

Plant Physiol.

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“…After killing, the leaves and leaf disks were exhaustively extracted with 80 The malic acid (COOH-CHOH-CH2-COOH) isolated was degraded by a variety of procedures. Carbon-4 was removed as CO, by Lactobacillus arabi- nosus in a modification of iNossal's method (19 (4 seconds). Furthermore, a similar distribution of label in malate was found after all other fixation periods.…”

Section: ) /8-carboxylase Activity Has Been Demonstratedmentioning

confidence: 99%

Malate Synthesis in Crassulacean Leaves. I. The Distribution of C¹⁴ in Malate of Leaves Exposed to C¹⁴O₂ in the Dark.

1958

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“…The age of culture had some effect on the course of "CO2 release from malic acid-"C (17). Cells of L. arabinosus harvested after 12, 24.…”

Section: Resultsmentioning

confidence: 99%

“…These authors used the L. arabinosus technique of degrading labeled malic acid. The degradative application of L. arabinosus was developed from the analytical applications described by Blanchard et al (4) and Nossal (16,17). Thus Bradbeer (5), Aronoff (1), and Avadhani (2) (18,19,21,27).…”

Section: Resultsmentioning

confidence: 99%

“…In all experiments, except those of Varner and Burrell (26), the labeling of the /3-carboxyl of malic acid was determined by means of malic acid-adapted Lactobacilltus arabinosus. This technique was developed from the observations of Blanchard et al (4) and was originally used as a manometric assay for malic acid by Nossal (16,17). The application of this technique to the degradation of labeled malic acid demands that the only significant fate of labeled malic acid fed to the organism is complete /3-decarboxylation via the endogenous NAD-malic enzyme.…”

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confidence: 99%

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Dark Fixation of CO₂ by Crassulacean Plants

Sutton¹,

Osmond²

1972

Plant Physiol.

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(11,27), poses the question of regulated carbon flow through the glycolytic and pentose phosphate pathways. As currently envisaged, the P-enolpyruvate consumed in dark CO2 fixation is derived from intermediates of the pentose phosphate pathway, rather than from glycolysis (25). The evidence for these events hinges on the observed distribution of "4C within malic acid-14C isolated from tissues showing CAM2. After short and long term "4CO2 fixation in the dark, Bradbeer et al. (6) reported that the distribution of label between the C-4 and C-1 carboxyl carbons of malic acid approximated to a constant ratio of 2: 1. Varner and Burrell (26) had previously reported an asymmetric distribution of label between the carboxyls of malic acid in Bryo-phyllutn calycinum and the approximate 2: 1 labeling pattern has been described in other succulent tissues in several subsequent reports (3, 9). Bradbeer et al. (6) interpreted this labeling pattern in terms of two carboxylation events, the first via ribu-'Supported by Commonwealth Postgraduate Research Award. 2 Abbreviation: CAM: Crassulacean acid metabolism. lose diphosphate carboxylase and the second via P-enolpyruvate carboxylase, as shown in Figure 1. Very little further support for this hypothesis has been advanced (21, 23).Labeling patterns in malic acid formed following preillumination and in dark fixation malic acid which approach 2:1 have been reported in other tissues (14,22). In all experiments, except those of Varner and Burrell (26), the labeling of the /3-carboxyl of malic acid was determined by means of malic acid-adapted Lactobacilltus arabinosus. This technique was developed from the observations of Blanchard et al. (4) and was originally used as a manometric assay for malic acid by Nossal (16,17). The application of this technique to the degradation of labeled malic acid demands that the only significant fate of labeled malic acid fed to the organism is complete /3-decarboxylation via the endogenous NAD-malic enzyme. The results presented here demonstrate that this is not always true. Using a more satisfactory in vitro degradation procedure, it is shown that labeled malic acid isolated from succulent tissues after dark '4CO2 fixation does not show a constant 2:1 labeling pattern. A preliminary report of these findings has been presented elsewhere (24).MATERIALS AND METHODS Growth of Plant Material. Stocks of Bryophyllum pinnatumn (Lamk.) Oken (B. calycinum Salisb.), Bryophyllum tubiflorum Harv., Kalanchoe daigremontiana Hamet et Perrier, and Sedum guatamalense Hemsl. were maintained in the glasshouse by vegetative propagation. The plants were grown in a sandy clay, adequately watered. Experimental plants were kept in a controlled environment chamber, 12 hr (25 C) light and 12 hr (17 C) dark. The humidity was kept constant at 65% relative humidity.Plants were used when about 3 months old. The phyllodes in the sixth to eighth whorl from the top of B. tubiflorum, the third pair of opposite leaves of K. daigremontiana, the second pair of opposite leaves of B. p...

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Estimation of l-malate and fumarate by malic decarboxylase of Lactobacillus arabinosus

Cited by 48 publications

References 11 publications

Malate Synthesis by Dark Carbon Dioxide Fixation in Leaves

Malate Synthesis by Dark Carbon Dioxide Fixation in Leaves

Malate Synthesis in Crassulacean Leaves. I. The Distribution of C¹⁴ in Malate of Leaves Exposed to C¹⁴O₂ in the Dark.

Dark Fixation of CO₂ by Crassulacean Plants

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Estimation of l-malate and fumarate by malic decarboxylase of Lactobacillus arabinosus

Cited by 48 publications

References 11 publications

Malate Synthesis by Dark Carbon Dioxide Fixation in Leaves

Malate Synthesis by Dark Carbon Dioxide Fixation in Leaves

Malate Synthesis in Crassulacean Leaves. I. The Distribution of C14 in Malate of Leaves Exposed to C14O2 in the Dark.

Dark Fixation of CO2 by Crassulacean Plants

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Malate Synthesis in Crassulacean Leaves. I. The Distribution of C¹⁴ in Malate of Leaves Exposed to C¹⁴O₂ in the Dark.

Dark Fixation of CO₂ by Crassulacean Plants