Studies on the lipid content and phosphate requirement of glucose- and acetate-grown <i>Escherichia coli</i>

Damoglou, A.P.; Dawes, E. A.

doi:10.1042/bj1100775

Cited by 32 publications

(20 citation statements)

References 7 publications

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“…In contrast, little or no change in lipid content of carbon-limited cultures was found for E. coli (Damoglou & Dawes, 1968;Calcott & Petty, 1980) or P . aeruginosa (Gilbert & Brown, 1978).…”

Section: Effects Of Growth Ratementioning

confidence: 73%

Phenotypic variation of lipid composition in Burkholderia cepacia: a response to increased growth temperature is a greater content of 2-hydroxy acids in phosphatidylethanolamine and ornithine amide lipid

1998

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Burkho/deria cepacia produces an unusual range of polar lipids, which includes two forms each of phosphatidylethanolamine (PE) and ornithine amide lipid (OL), differing in the presence or absence of 2-hydroxy fatty acids. By using chemostat cultures in chemically defined media, variations in the lipid content and the proportions of individual lipids have been studied as a function of (a) growth temperature, (b) growth rate and (c) growth-limiting nutrient (carbon, magnesium, phosphorus or oxygen). Total cellular lipid in carbon-limited cultures was lowest at high growth temperatures and low growth rates. Increases in growth temperature over the range 2540 O C led t o increases in the proportions of molecular species of PE and OL containing 2-hydroxy acids, without changing the PE:OL ratio. Growth temperature did not alter the balance between neutral and acidic lipids, but the contribution of phosphatidylglycerol to the latter increased with rising growth temperature and growth rate. Pigmentation of cells and the presence of flagella were also temperature-dependent. Change in growth rate also affected the PE :OL ratio and the extent to which monoenoic acids were replaced by their cyclopropane derivatives. Whereas similar lipid profiles were found for carbon-, magnesiumand oxygen-limited cultures, ornithine amides were the only polar lipids detected in phosphorus-limited cells.

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Section: Effects Of Growth Ratementioning

confidence: 73%

Phenotypic variation of lipid composition in Burkholderia cepacia: a response to increased growth temperature is a greater content of 2-hydroxy acids in phosphatidylethanolamine and ornithine amide lipid

1998

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“…S1). This spectral pattern reflects a substitution of C-H X by C-D X in newly synthesized macromolecules in the presence of heavy water, and it is safe to assume that an important fraction of the D incorporation is found in lipids, which contribute roughly 25-30% to total cellular C-H (45,46). Lipids from autotrophic and heterotrophic organisms show a predominance of water-derived protons (or D + in the presence of heavy water) (33,35,47) as protons from water are transferred to NAD(P) during its reduction, and these protons are then incorporated into lipids via the known fatty acid biosynthesis pathways (31)(32)(33).…”

Section: Resultsmentioning

confidence: 99%

Tracking heavy water (D ₂ O) incorporation for identifying and sorting active microbial cells

Berry

Mader

Lee

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

369

512

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Microbial communities are essential to the function of virtually all ecosystems and eukaryotes, including humans. However, it is still a major challenge to identify microbial cells active under natural conditions in complex systems. In this study, we developed a new method to identify and sort active microbes on the single-cell level in complex samples using stable isotope probing with heavy water (D 2 O) combined with Raman microspectroscopy. Incorporation of D 2 O-derived D into the biomass of autotrophic and heterotrophic bacteria and archaea could be unambiguously detected via C-D signature peaks in single-cell Raman spectra, and the obtained labeling pattern was confirmed by nanoscaleresolution secondary ion MS. In fast-growing Escherichia coli cells, label detection was already possible after 20 min. For functional analyses of microbial communities, the detection of D incorporation from D 2 O in individual microbial cells via Raman microspectroscopy can be directly combined with FISH for the identification of active microbes. Applying this approach to mouse cecal microbiota revealed that the host-compound foragers Akkermansia muciniphila and Bacteroides acidifaciens exhibited distinctive response patterns to amendments of mucin and sugars. By Ramanbased cell sorting of active (deuterated) cells with optical tweezers and subsequent multiple displacement amplification and DNA sequencing, novel cecal microbes stimulated by mucin and/ or glucosamine were identified, demonstrating the potential of the nondestructive D 2 O-Raman approach for targeted sorting of microbial cells with defined functional properties for singlecell genomics.ecophysiology | single-cell microbiology | carbohydrate utilization | nitrifier | Raman microspectroscopy M icroorganisms play a vital role in many environments. They mediate global biogeochemical cycles, catalyze biotechnological processes, and contribute to health and disease in the human body. The in situ study of microbial activity in natural and engineered ecosystems is therefore of great interest. For this purpose, several elegant methods have been established that use either transcriptional or translational activity of community members (i.e., metatranscriptomics, metaproteomics) (1-3) or the incorporation of isotopically labeled substrates into biomolecules (4-10) to infer the ecophysiology of microbes in such systems. However, these bulk techniques do not offer sufficient spatial resolution to study microbial activities at the micrometer scale. Therefore, important information can be overlooked because microbial communities are frequently spatially structured (e.g., biofilms) (11) and contain populations with life cycles (12,13). Furthermore, even apparently identical cells in clonal populations can have strongly divergent activities (14).Consequently, microbial ecophysiology is ideally studied also at the level of the single cell, but only a restricted number of approaches exist for determining physiological properties of individual cells in a microbial community. For exa...

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“…type 6097s) with a Nuclear Enterprises type N.E. 5353 high voltage supply (Damoglou & Dawes, 1968). The current output of the tube was measured after conversion to voltage on a Yellow Springs Model 80 laboratory recorder (Yellow Springs Instrument Co. Inc., Yellow Springs, Ohio, U.S.A.).…”

Section: Methodsmentioning

confidence: 99%

Effect of Starvation on the Viability and Cellular Constituents of Zymomonas anaerobia and Zymomonas mobilis

Dawes

Large

1970

Journal of General Microbiology

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SUMMARYZymomonas anaerobia and 2. mobilis, grown on glucose + peptone + yeastextract medium, degraded no endocellular carbohydrate, DNA and protein even in prolonged starvation. No significant qualitative changes in protein content during starvation were detected by disc gel electrophoresis of crude extracts. Both organisms had a high content of RNA (22 % w/w) which was degraded on starvation. In 2. anaerobia RNA decreased linearly to 5 % of the dry weight in 125 hr. With 2. mobilis, half the RNA was degraded in the first 24 hr of starvation after which time the decline was much slower. MgC12 (33 mM) prevented RNA breakdown. During growth, the intracellular ATP concentration increased from 0.5 to 1.0 ,ug./mg. dry wt, but began to decrease exponentially in the last generation before growth ceased because of glucose exhaustion. Intracellular ATP content correlated with viability determined by slide culture. The addition of 33 mM-MgC1, to the starvation medium did not affect ATP content, but increased viability. On prolonged starvation (up to 7 days), populations whose viability had fallen to 3 % possessed unimpaired ability to produce ATP from glucose; only after even longer starvation periods was this ability impaired. I N T R O D U C T I O NDuring recent years considerable attention has been focused on the nature of the endogenous metabolism of micro-organisms (for reviews, see Dawes & Ribbons, 1962, 1964 although this work has been confined mainly to aerobic and facultatively anaerobic organisms. By investigating the changes in chemical composition which occur during starvation an insight can be afforded into the substrates utilized in endogenous metabolism and their relation to the viability of the organisms. Anaerobic bacteria, which obtain their energy by substrate-level phosphorylation reactions which are well charted, ought to be particularly amenable to investigation, especially in relation to the problem of 'maintenance energy' (Dawes & Ribbons, 1964 (NCIB 8938) was maintained in a similar manner. This organism grew with a doubling time of about 2 hr, compared with 3.5 hr for 2. anaerobia.Growth of larger quantities of either organism was achieved by transfer of an inoculum from the stock culture to 20 ml. sterile liquid medium. When this culture was gassing vigorously (after about 12 hr) it was used to inoculate one or more litres of medium in round flat-bottomed flasks or aspirators filled to the neck. Zymomonas anaerobia attained stationary phase in 20 hr at 30°, while 2. mobilis took 12 to 15 hr.For starvation experiments organisms were harvested aseptically in closed, sterile centrifuge bottles and the cultures were examined for contamination by phasecontrast microscopic examination of a wet smear.Measurement of bacterial concentration. This was performed turbidimetrically with a Unicam SP 600 spectrophotometer at 570 nm. Extinctions were related to population density by a dry wt calibration curve.Preparation of washed suspensions for starvation experiments. After the appropriate period of growth ...

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Studies on the lipid content and phosphate requirement of glucose- and acetate-grown Escherichia coli

Cited by 32 publications

References 7 publications

Phenotypic variation of lipid composition in Burkholderia cepacia: a response to increased growth temperature is a greater content of 2-hydroxy acids in phosphatidylethanolamine and ornithine amide lipid

Phenotypic variation of lipid composition in Burkholderia cepacia: a response to increased growth temperature is a greater content of 2-hydroxy acids in phosphatidylethanolamine and ornithine amide lipid

Tracking heavy water (D ₂ O) incorporation for identifying and sorting active microbial cells

Effect of Starvation on the Viability and Cellular Constituents of Zymomonas anaerobia and Zymomonas mobilis

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Studies on the lipid content and phosphate requirement of glucose- and acetate-grown Escherichia coli

Cited by 32 publications

References 7 publications

Phenotypic variation of lipid composition in Burkholderia cepacia: a response to increased growth temperature is a greater content of 2-hydroxy acids in phosphatidylethanolamine and ornithine amide lipid

Phenotypic variation of lipid composition in Burkholderia cepacia: a response to increased growth temperature is a greater content of 2-hydroxy acids in phosphatidylethanolamine and ornithine amide lipid

Tracking heavy water (D 2 O) incorporation for identifying and sorting active microbial cells

Effect of Starvation on the Viability and Cellular Constituents of Zymomonas anaerobia and Zymomonas mobilis

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Tracking heavy water (D ₂ O) incorporation for identifying and sorting active microbial cells