An improved synthetic growth medium for <i>Halobacterium cutirubrum</i>

Grey, Vijaylaxmi; Fitt, Peter S.

doi:10.1139/m76-068

Cited by 42 publications

(20 citation statements)

References 2 publications

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“…Cultures were grown at 37°C with shaking for 4 days to stationary phase. Cells used as inoculum were harvested by centrifugation (10,000 ϫ g for 2 min using a Beckman-Coulter [Fullerton, Calif.] Microfuge 18 centrifuge) and washed once with chemically defined Grey and Fitt medium, pH 6.6 (13). Cells were diluted 100-fold and used to inoculate the defined medium containing the appropriate corrinoid supplements.…”

Section: Methodsmentioning

confidence: 99%

ThecobYGene of the ArchaeonHalobacteriumsp. Strain NRC-1 Is Required for De Novo Cobamide Synthesis

Woodson

Peck²,

Krebs

et al. 2003

J Bacteriol

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Genetic and nutritional analyses of mutants of the extremely halophilic archaeon Halobacterium sp. strain NRC-1 showed that open reading frame (ORF) Vng1581C encodes a protein with nucleoside triphosphate: adenosylcobinamide-phosphate nucleotidyltransferase enzyme activity. This activity was previously associated with the cobY gene of the methanogenic archaeon Methanobacterium thermoautotrophicum strain ⌬H, but no evidence was obtained to demonstrate the direct involvement of this protein in cobamide biosynthesis in archaea. Computer analysis of the Halobacterium sp. strain NRC-1 ORF Vng1581C gene and the cobY gene of M. thermoautotrophicum strain ⌬H showed the primary amino acid sequence of the proteins encoded by these two genes to be 35% identical and 48% similar. A strain of Halobacterium sp. strain NRC-1 carrying a null allele of the cobY gene was auxotrophic for cobinamide-GDP, a known intermediate of the late steps of cobamide biosynthesis. The auxotrophic requirement for cobinamide-GDP was corrected when a wild-type allele of cobY was introduced into the mutant strain, demonstrating that the lack of cobY function was solely responsible for the observed block in cobamide biosynthesis in this archaeon. The data also show that Halobacterium sp. strain NRC-1 possesses a high-affinity transport system for corrinoids and that this archaeon can synthesize cobamides de novo under aerobic growth conditions. To the best of our knowledge this is the first genetic and nutritional analysis of cobalamin biosynthetic mutants in archaea.

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Section: Methodsmentioning

confidence: 99%

ThecobYGene of the ArchaeonHalobacteriumsp. Strain NRC-1 Is Required for De Novo Cobamide Synthesis

Woodson

Peck²,

Krebs

et al. 2003

J Bacteriol

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“…Unless specified otherwise, strains were grown at 37 O C in peptone medium (Lanyi e t al., 1979) in the dark with vigorous agitation. Defined medium was prepared as described by Grey & Fitt (1976); basal salts buffer was prepared as for peptone medium except for the omission of Oxoid peptone and the inclusion of 10 mM MES, pH 6.8. Behavioural assays.…”

Section: Methodsmentioning

confidence: 99%

Aerotaxis in Halobacterium salinarium is methylation-dependent

Lindbeck

Goulbourne²,

Johnson³

et al. 1995

Microbiology

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The behavioural response to a gradient of oxygen (aerotaxis) has been characterized in the archaeon, Halobecterium salinarium. When the gas surrounding a drop of H. saharium strain S9-P culture was changed abruptly from 10% (vh) 0, to 100% N , , the bacteria transiently increased the frequency of reversing before they adapted and resumed random swimming. When the gas was returned to 10% 0, the bacteria responded by swimming smoothly for approximately 45 s. Aerotaxis was strongest when respiration in H. salinarium was highest and when bacteriorhodopsin and halorhodopsin were not contributing to the proton motive force. Starvation for methionine of the auxotrophic H . salinarium essentially abolished the step-down aerotactic response. Methanol production from demethylation of methyl-accepting chemotaxis proteins was transiently increased in H . salinarium S9-P by a step down or step up in oxygen concentration, as observed in methylationdependent chemotaxis in H. salinarium. The taxislnegative and methyltransferase-deficient mutant, H . salinarium strain Pho72 did not exhibit changes in methanol release in response to aerotaxis or chemotaxis stimuli. This is the first report of an aerotactic response that is dependent on methylation of methyl-accepting chemotaxis proteins. Aerotaxis in Escherichia coli and Salmonella typhimurium is independent of transducer methylation.

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“…H. cutirubrum (NRC 34001) and H. halobium (NRC 34003) were grown aerobically in the dark at 37°C in 100-ml volumes or in 50-ml volumes in experiments with '3C-labeled lysine. The synthetic growth medium described by Grey and Fitt (12) was used, except in amino acid biosynthesis experiments, where the amino acid being studied was omitted from the medium. Sodium citrate, originally present in the medium (12) For '4C labeling, 5 ,uCi of each substrate was added to 100 ml of medium containing the full complement of amino acids, glycerol (100 mg/100 ml), and CH3COONa.…”

Section: Methodsmentioning

confidence: 99%

Mevalonic acid is partially synthesized from amino acids in Halobacterium cutirubrum: a 13C nuclear magnetic resonance study

Ekiel¹,

Sprott²,

Smith³

1986

J Bacteriol

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13C nuclear magnetic resonance revealed an unusual pathway for the biosynthesis of lipids in Halobacterium cutirubrum and H. halobium. Mevalonic acid was not synthesized from three acetyl-coenzyme A molecules, as has been suggested previously, and the branch-methyl and methine carbons in phytanyl chains were derived from neither acetate nor glycerol. Instead, they were supplied by the degradation of amino acids, in particular of lysine. Presumably, two different types of two-carbon fragments were used simultaneously by halobacteria for the biosynthesis of mevalonate. The labeling pattern of squalene supported the above conclusions. Based on these data, a general scheme is proposed to account for the contribution of lysine-to-lipid biosynthesis.Our previous papers on the application of the 13C-nuclear magnetic resonance (NMR) technique to study the metabolic pathways of methanogenic bacteria (7,8,10) showed several significant differences in biosynthesis when compared with the biosynthesis of other amino acids by bacteria and eucaryotes. Similar studies for comparison with halobacteria are lacking. Halobacteria are archaebacteria also, but with an aerobic rather than anaerobic mode of metabolism.Halobacteria require quite complicated media for growth, and, in contrast to the aceticlastic methanogens, acetate can be used for growth only when combined with several additional carbon sources.

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An improved synthetic growth medium for Halobacterium cutirubrum

Abstract: A synthetic medium is described in which Halobacterium cutirubrum grows as well as in complex media.

Cited by 42 publications

References 2 publications

ThecobYGene of the ArchaeonHalobacteriumsp. Strain NRC-1 Is Required for De Novo Cobamide Synthesis

ThecobYGene of the ArchaeonHalobacteriumsp. Strain NRC-1 Is Required for De Novo Cobamide Synthesis

Aerotaxis in Halobacterium salinarium is methylation-dependent

Mevalonic acid is partially synthesized from amino acids in Halobacterium cutirubrum: a 13C nuclear magnetic resonance study

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