The malate dehydrogenase of Ralstonia eutropha and functionality of the C3/C4 metabolism in a Tn5-induced mdh mutant

Brämer, Christian O

doi:10.1016/s0378-1097(02)00745-0

Cited by 8 publications

(23 citation statements)

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“…We decided to use trioleate as a representative triacylglycerol, as trioleate is a uniform, defined carbon source, as opposed to plant oils, which may contain contaminating compounds that could add unwanted complexity to the analysis of the microarray data. We monitored the concentration of NH 4 Cl in the cultures so that samples from each culture were taken at approximately the same phase of growth. To represent the logarithmic growth phase, we took samples when cultures had utilized approximately half of the NH 4 Cl in the medium (ϳ250 g/ml).…”

Section: Resultsmentioning

confidence: 99%

“…We monitored the concentration of NH 4 Cl in the cultures so that samples from each culture were taken at approximately the same phase of growth. To represent the logarithmic growth phase, we took samples when cultures had utilized approximately half of the NH 4 Cl in the medium (ϳ250 g/ml). Samples were also taken ϳ2 h after all NH 4 Cl in the culture was depleted, representing the PHB production phase.…”

Section: Resultsmentioning

confidence: 99%

“…Four individual colonies of R. eutropha H16 grown on a tryptic soy agar (TSA) plate were inoculated into 5 ml of dextrose-free tryptic soy broth (TSB; Becton Dickinson, Sparks, MD) and grown for 24 h. Aliquots of 0.5 ml of overnight culture were inoculated into 250-ml shake flasks containing 50 ml of minimal medium, modified from reference 36, containing 0.1% NH 4 Cl and either 2% (wt/vol) fructose or 1% (wt/vol) trioleate, emulsified with 0.3% (wt/vol) gum arabic. These cultures were grown for 24 h. Overnight cultures were inoculated to an initial optical density at 600 nm (OD 600 ) of 0.1 into 250-ml shake flasks containing 50 ml of minimal medium containing 0.05% NH 4 Cl and either 2% (wt/vol) fructose or 1% (wt/vol) trioleate, emulsified with 0.3% (wt/vol) gum arabic. Cultures were grown for 12 h. Cultures for sampling were inoculated to an initial OD 600 of 0.05 in 250-ml shake flasks containing 50 ml of minimal medium with 0.05% NH 4 Cl and either 2% (wt/vol) fructose or 1% (wt/vol) trioleate, emulsified with 0.3% (wt/vol) gum arabic.…”

Section: Methodsmentioning

confidence: 99%

“…These cultures were grown for 24 h. Overnight cultures were inoculated to an initial optical density at 600 nm (OD 600 ) of 0.1 into 250-ml shake flasks containing 50 ml of minimal medium containing 0.05% NH 4 Cl and either 2% (wt/vol) fructose or 1% (wt/vol) trioleate, emulsified with 0.3% (wt/vol) gum arabic. Cultures were grown for 12 h. Cultures for sampling were inoculated to an initial OD 600 of 0.05 in 250-ml shake flasks containing 50 ml of minimal medium with 0.05% NH 4 Cl and either 2% (wt/vol) fructose or 1% (wt/vol) trioleate, emulsified with 0.3% (wt/vol) gum arabic. All flask cultures were grown at 30°C with agitation (200 rpm).…”

Section: Methodsmentioning

confidence: 99%

“…Overnight cultures were washed and diluted 1:10 in sterile saline. Aliquots of 50 l of a 1:10 dilution of overnight culture were inoculated into 250-ml shake flasks containing 50 ml of minimal medium, modified from reference 36, containing 0.1% NH 4 Cl and 1% (wt/vol) palm oil, CPKO, or oleic acid, emulsified with 0.3% (wt/vol) gum arabic. These cultures were grown for up to 72 h at 30°C with agitation (200 rpm).…”

Section: Construction Of Complementation Plasmids and Introduction Inmentioning

confidence: 99%

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Elucidation of β-Oxidation Pathways in Ralstonia eutropha H16 by Examination of Global Gene Expression

Brigham¹,

Budde²,

Holder³

et al. 2010

J Bacteriol

110

109

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Ralstonia eutropha H16 is capable of growth and polyhydroxyalkanoate production on plant oils and fatty acids. However, little is known about the triacylglycerol and fatty acid degradation pathways of this bacterium. We compare whole-cell gene expression levels of R. eutropha H16 during growth and polyhydroxyalkanoate production on trioleate and fructose. Trioleate is a triacylglycerol that serves as a model for plant oils. Among the genes of note, two potential fatty acid ␤-oxidation operons and two putative lipase genes were shown to be upregulated in trioleate cultures. The genes of the glyoxylate bypass also exhibit increased expression during growth on trioleate. We observed that single ␤-oxidation operon deletion mutants of R. eutropha could grow using palm oil or crude palm kernel oil as the sole carbon source, regardless of which operon was present in the genome, but a double mutant was unable to grow under these conditions. A lipase deletion mutant did not exhibit a growth defect in emulsified oil cultures but did exhibit a phenotype in cultures containing nonemulsified oil. Mutants of the glyoxylate shunt gene for isocitrate lyase were able to grow in the presence of oils, while a malate synthase (aceB) deletion mutant grew more slowly than wild type. Gene expression under polyhydroxyalkanoate storage conditions was also examined. Many findings of this analysis confirm results from previous studies by our group and others. This work represents the first examination of global gene expression involving triacylglycerol and fatty acid catabolism genes in R. eutropha.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Construction Of Complementation Plasmids and Introduction Inmentioning

confidence: 99%

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Elucidation of β-Oxidation Pathways in Ralstonia eutropha H16 by Examination of Global Gene Expression

Brigham¹,

Budde²,

Holder³

et al. 2010

J Bacteriol

110

109

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CC Bond-Forming Reactions of IrIII-Alkenyls and Nitriles or Aldehydes: Generation of Reactive Hydride- and Alkyl-Alkylidene Compounds and Observation of a Reversible 1,2-H Shift in Stable Hydride–IrIII Alkylidene Complexes

et al. 2002

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Reaction kinetics of polyureas with acetylated diamine chain extenders

Song

Liu

2011

J of Applied Polymer Sci

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Diacetyl hexamethylene-diamine (MHAD) and diacetyl m-phenylenediamine (Mm-PDA) were synthesized by acetylating hexamethyene-diamin (HAD) and mphenylenediamine (m-PDA) with glacial acetic acid in presence of phosphoric acid, in order to retard the high reactivity of polyurea. Chemical structures of modified diamine chain extenders were confirmed through FTIR, the determination of the nitrogen content, High resolution ESI-Fourier transform ion cyclotron resonance (ESI-FTICR) mass spectrometry and 1 H-NMR analysis. Polyureas were synthesized via two-step solution process by reaction of 4,4 0 -diphenyl diisocyanate (MDI), amine terminated polyether (Jeffamine D-2000) with different chain extenders. The influence of modified diamine chain extenders on gel time of polyurea was investigated. The gel time was prolonged from 11s to 52s and 13s to 65s respectively. The reaction kinetics of synthesizing polyureas were studied by FTIR. The results showed that extending reaction with MHAD was second order kinetic mechanism, its apparent activation energy was 43.99 kJÁmol À1 ; extending reaction with Mm-PDA reaction was first order, its activation energy was 58.95 kJÁmol À1 .

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The malate dehydrogenase of Ralstonia eutropha and functionality of the C3/C4 metabolism in a Tn5-induced mdh mutant

Cited by 8 publications

References 0 publications

Elucidation of β-Oxidation Pathways in Ralstonia eutropha H16 by Examination of Global Gene Expression

Elucidation of β-Oxidation Pathways in Ralstonia eutropha H16 by Examination of Global Gene Expression

CC Bond-Forming Reactions of IrIII-Alkenyls and Nitriles or Aldehydes: Generation of Reactive Hydride- and Alkyl-Alkylidene Compounds and Observation of a Reversible 1,2-H Shift in Stable Hydride–IrIII Alkylidene Complexes

Reaction kinetics of polyureas with acetylated diamine chain extenders

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