2013
DOI: 10.1016/j.mimet.2013.02.006
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An accurate method for estimation of the intracellular aqueous volume of Escherichia coli cells

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Cited by 16 publications
(17 citation statements)
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“…1. First, wabG, which encodes a glucosyltransferase that plays a key role in the synthesis of the outer core LPS, was removed to reduce the strain's pathogenicity (29), and the resulting strain was named KMK-01. LPS, the outer component of the bacterial surface, is considered an important pathogenic determinant in K. pneumoniae.…”
Section: Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…1. First, wabG, which encodes a glucosyltransferase that plays a key role in the synthesis of the outer core LPS, was removed to reduce the strain's pathogenicity (29), and the resulting strain was named KMK-01. LPS, the outer component of the bacterial surface, is considered an important pathogenic determinant in K. pneumoniae.…”
Section: Resultsmentioning
confidence: 99%
“…To construct the wabG deletion mutant, K. pneumoniae KCTC 2242 containing pRedET was cultured at 30°C, and 10% L-arabinose in deionized water was added to a final concentration of 24 mM when the optical density at 600 nm (OD 600 ) reached 0.2. A kanamycin resistance gene flanked with Flp recombination target (FRT) sites was amplified from pKD4 with the primers wabG-FKF-fw and to 1.9 l/mg (29). The experiment was repeated three times independently.…”
Section: Methodsmentioning
confidence: 99%
“…However, one uncertainty affecting the data has to be considered: For the calculation of intracellular metabolite concentrations the intracellular aqueous volume of an E. coli cell is required. Here, a volume of 1.9 mL g -1 determined by Wang et al (2013) for E. coli BL21(DE3) was used. Other references, however, describe higher values of 2.15-3.2 mL g -1 , with the values highly varying depending on the overall cell fitness, the carbon source, the growth rate and the type of medium (Bennett et al 2008;Park et al 2011;Volkmer and Heinemann 2011).…”
Section: Discussionmentioning
confidence: 99%
“…In order to calculate the intracellular concentration, cell dry weights were determined in reactors operated under identical conditions as the sampled reactors. With an assumed intracellular aqueous volume of 1.9 mL g -1 for E. coli BL21(DE3), as proposed by Wang et al (2013), the total cellular volume V X in one sample was calculated. By considering the dilution of the aqueous reaction volume by the triethanolamine buffer (20 mL), the intracellular concentration c A,intra was estimated:…”
Section: Quantification Of Intracellular Udp-glucose and Udpmentioning
confidence: 99%
“…E. coli dry cell weight (DCW) was weight by converting OD 600 value with a coefficient of 0.275 g DCW /(L × OD 600 ), which was determined by freezer drying the E. coli cells according to our recently report [ 36 ]. As the DHA production was performed at 0.1 M potassium phosphate buffer (pH 9.0) due to GldA activity has higher activity as mentioned above, all enzymatic assays were performed at consistent pH of 9.0.…”
Section: Methodsmentioning
confidence: 99%