Mutants of Escherichia coli K12 unable to grow on non-fermentable carbon substrates

Daniel, Jacques; Roisin, Marie‐Paule; Burstein, Claude; Képès, Adam

doi:10.1016/0005-2728(75)90011-0

Cited by 23 publications

(9 citation statements)

References 25 publications

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“…Finally, neither intact or washed membranes from mutant N144 could be reactivated by the addition of parental F1, an observation which suggested that this mutation had produced a catalytically defective F1 that was more firmly bound to the membrane than the wild-type enzyme (301). The MDA1, MDA2, and MDA3 mutants isolated by Kepes and his colleagues exhibited ATPase-binding and reactivation properties which suggested very close similarities with the uncA, unc-405, and N144 mutants, respectively (93).…”

Section: Mutants Defective In Energy Transductionmentioning

confidence: 99%

Bacterial respiration.

Haddock¹,

Jones²

1977

Bacteriological Reviews

392

205

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Section: Mutants Defective In Energy Transductionmentioning

confidence: 99%

Bacterial respiration.

Haddock¹,

Jones²

1977

Bacteriological Reviews

392

205

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“…The mutants MDA (BFI ATPase negative) and MDB which is BF1 ATPase positive (dicyclohexylcarbodiimide-resistant) are devoid of membranebound ATPase [14] and present no respiratory stimulation under the experimental conditions here described.…”

Section: Effect Of Different Mutations On the Respiratory Controlmentioning

confidence: 99%

“…Two other types of mutants, MDAz and MDA3, which have an altered BFI ATPase still bound to their membrane but inactive [14] also are surprisingly unable to exhibit respiratory stimulation by CCCP. This loss of respiratory control is in correlation with the loss of the oxygen-driven transhydrogenase [15].…”

Section: Effect Of Different Mutations On the Respiratory Controlmentioning

confidence: 99%

Respiratory Control in Escherichia coli K 12

Burstein

Tiankova

Képès

1979

European Journal of Biochemistry

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Contrary to the general view point that respiratory control does not exist in Escherichia coli, we have been able to demonstrate this phenomenon in this micro-organism. E. coli cells growing aerobically on an oxidizable carbon source respire at a high rate, which cannot be increased by the addition of uncouplers of oxidative phosphorylation. Upon removal of the carbon source, the rate of respiration decreases progressively to reach, after 10 min, less than 1/20 of its previous value. Under these conditions the addition of an uncoupler increases the residual respiration by a factor of 3 to 9. This increase is due to the release of respiratory control. Release of respiratory control cannot be observed upon addition of ADP which does not penetrate across the membrane. Respiratory control can also be shown in the presence of non-limiting respiratory substrate provided that it is not a carbon source. This was shown by measuring the rate of oxidation of ascorbate and phenazine methosulfate.NADH oxidase activity of membrane vesicles obtained by high pressure disintegration was increased by a factor of about 1.5 upon the addition of uncouplers and also upon addition of ADP plus phosphate. In contrast, membrane vesicles depleted from BF1 ATPase (a soluble bacterial coupling factor presenting ATPase activity) did not exhibit respiratory control. It could be restored either by rebinding of BF1 ATPase or by addition of dicyclohexylcarbodiimide, an ATPase inhibitor.The limited respiratory rate of mitochondria during so called 'state 4' which can be increased by uncouplers or by ADP plus phosphate is attributed to respiratory control [l].The lack of respiratory stimulation by the same agents on respiring Escherichia coli led several investigators [2--41 to state that there was no respiratory control in E. coli. It appears that growing E. coli cells respire at maximal rate, presumably because ATP utilising synthetic processes continuously liberate ADP plus phosphate. The increase in respiration of starved cells observed upon addition of an actively transported lactose analogue, methyl p-Dthiogalactoside was interpreted also as a release of respiratory control by the utilization of metabolic energy [5]. In this paper, we describe conditions which allow the demonstration of respiratory control in intact E. coli without added carbon source by its release upon addition of uncouplers. That this is not due to limiting oxidizable substrate can be shown by adding excess ascorbate and phenazine methosulfate.Ahhreviations. CCCP, carbonylcyanide-p-trichloromethoxyphenylhydrazone; BF1 ATPase: soluble bacterial coupling factor F1 presenting ATPase activity.Addition of ADP, a non-penetrating solute, does not cause the same respiratory increase. We also describe a less pronounced increase of respiration, with NADH as the reducing substrate in a cell-free membrane preparation which responds to the addition of uncouplers as well as of ADP plus Pi. MATERIALS AND METHODSGrowth of Bacteria E. coli K12 strain 3300 constitutive for the lactose oper...

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“…In contrast, nitrogen-fixing Rhizobium bacteroids apparently cannot utilize glucose (Burris and Wilson 1939;Tuzimura and Megura 1960;de Vries and Hordijk 1978), but can use TCA-cycle intermediates such as succinate and fumarate which are supplied by the plant. Sufficient ATP cannot be derived from such compounds by substrate level phosphorylation (Harold 1972;Daniel et al 1975). In addition, the bacteroids are constantly supplied with oxygen by leghaemoglobin (Appleby et al 1975).…”

Section: Discussionmentioning

confidence: 99%

“…Growth of NmRSuc-mutants of R. tri/olii on various carbon sources Growth was assessed on plates of TM medium with the various compounds as sole carbon source, present at 20 mM (except for malate and D-lactate which were present at 2 mM, since 20 mM inhibits the growth of strain Tl). Plants were incubated aerobically for 5 days at 30 o e. Growth response: -, no growth; +, + +, + + +, + + + + ; increasing degrees of growth from poor to very good, assessed both in regions of confluent growth and on size of isolated colonies (Cox and Gibson 1974;Daniel et al 1975;Thipayathasana 1975), but can grow on glucose or on glycerol, since energy can be derived from these compounds by substrate level phosphorylation (Daniel et al 1975). Since all R. trifolii NmRSuc-strains had proved to be completely unable to use succinate, they were tested for growth in TM media with other TCA cycle intermediates and related compounds as sole carbon source.…”

Section: Properties 0/ Nmflsuc -Mutantsmentioning

confidence: 99%

Bacterial and Bacteroid Properties of Mutants of Rhizobium Trifolii Strain T1 Uncoupled in Oxidative Phosphorylation

Skotnicki¹,

Rolfe²

1979

Aust. Jnl. Of Bio. Sci.

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Using the neomycin-resistance selection technique, mutants uncoupled in oxidative phosphorylation were isolated in R. tri/olii strain Tl. These mutants had no detectable ATP-synthesizing activity and between 8 and 20% of the ATP-degrading activity of their parent strain in the bacterial form. The mutants formed nitrogen-fixing nodules on red, white, and subterranean clovers, and bacteroids isolated from white clover nodules had a high level of ATPase activity of about 70% of that found in bacteroids of their parental strain. The possibility that a new ATPase complex is synthesized when the bacteria differentiate to form bacteroids within the nodule is discussed.

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Mutants of Escherichia coli K12 unable to grow on non-fermentable carbon substrates

Cited by 23 publications

References 25 publications

Bacterial respiration.

Bacterial respiration.

Respiratory Control in Escherichia coli K 12

Bacterial and Bacteroid Properties of Mutants of Rhizobium Trifolii Strain T1 Uncoupled in Oxidative Phosphorylation

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