Determination of the maximum specific uptake capacities for glucose and oxygen in glucose‐limited fed‐batch cultivations of <i>Escherichia coli</i>

Lin, Hongying; Mathiszik, B.; Xu, Bo; Enfors, Sven‐Olof; Neubauer, Peter

doi:10.1002/bit.1068

Cited by 85 publications

(95 citation statements)

References 33 publications

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“…The average cellular respiration rate is calculated by normalizing OUR to population size. Most studies of this nature determine respiration rates while cells are growing (9)(10)(11)(12)(13)(14). In the rare cases where respiration was measured after cells exited exponential-phase growth (15,16), respiration rates dropped significantly-presumably as the cells became increasingly nutrient limited.…”

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Oxygen Consumption Rates of Bacteria under Nutrient-Limited Conditions

Riedel

Berelson

Nealson

et al. 2013

Appl Environ Microbiol

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bMany environments on Earth experience nutrient limitation and as a result have nongrowing or very slowly growing bacterial populations. To better understand bacterial respiration under environmentally relevant conditions, the effect of nutrient limitation on respiration rates of heterotrophic bacteria was measured. The oxygen consumption and population density of batch cultures of Escherichia coli K-12, Shewanella oneidensis MR-1, and Marinobacter aquaeolei VT8 were tracked for up to 200 days. The oxygen consumption per CFU (Q O2 ) declined by more than 2 orders of magnitude for all three strains as they transitioned from nutrient-abundant log-phase growth to the nutrient-limited early stationary phase. The large reduction in Q O2 from growth to stationary phase suggests that nutrient availability is an important factor in considering environmental respiration rates. Following the death phase, during the long-term stationary phase (LTSP), Q O2 values of the surviving population increased with time and more cells were respiring than formed colonies. Within the respiring population, a subpopulation of highly respiring cells increased in abundance with time. Apparently, as cells enter LTSP, there is a viable but not culturable population whose bulk community and per cell respiration rates are dynamic. This result has a bearing on how minimal energy requirements are met, especially in nutrient-limited environments. The minimal Q O2 rates support the extension of Kleiber's law to the mass of a bacterium (100-fg range). Heterotrophic bacterial respiration constitutes 50% to 90% of ocean community respiration (1) and plays a critical role in the recycling of organic carbon in all natural environments (2). Despite this importance, respiration remains poorly quantified in ocean models of metabolism, gas exchange, and carbon mass balances (3). This uncertainty in heterotrophy makes it difficult to predict when areas of the ocean will be sources or sinks of CO 2 (4). Microbial community ecosystem behavior is ultimately a function of auto-and heterotrophic relationships; hence, understanding the biogeochemical contribution of heterotrophic bacteria at the single-cell level is necessary for making predictions of how ecosystems will respond to global change (5). Extracting this information is difficult. Efforts are being made with single-cell observatories designed to trap cells in microwells and monitor their consumption of oxygen over time using optical techniques (6, 7). Field-based approaches involve modeling oxygen pore water profiles in marine sediments and dividing oxygen consumption rate by cell numbers (8). A more frequently employed method for analyzing cellular respiration rates is to determine bulk oxygen uptake (consumption) rates (OUR), using either batch or continuous cultures with large populations of microbes. The average cellular respiration rate is calculated by normalizing OUR to population size. Most studies of this nature determine respiration rates while cells are growing (9-14). In the rare cases wher...

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confidence: 99%

Oxygen Consumption Rates of Bacteria under Nutrient-Limited Conditions

Riedel

Berelson

Nealson

et al. 2013

Appl Environ Microbiol

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“…In the literature, several specific fermentation strategies are mentioned to reduce acetate production [1,[16][17][18][19][20]. levels were developed.…”

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Minimizing acetate formation in E. coli fermentations

Mey

Maeseneire

Soetaert

et al. 2007

J Ind Microbiol Biotechnol

199

144

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Escherichia coli remains the best established production organisms in industrial biotechnology. However, during aerobic fermentation runs at high growth rates, considerable amounts of acetate are accumulated as by-product. This by-product has negative effects on growth and protein production. Over the last 20 years, substantial research efforts have been spent to reduce acetate accumulation during aerobic growth of E. coli on glucose. From the onset it was clear that this quest should not be a simple nor uncomplicated one. Simple deletion of the acetate pathway, reduced the acetate accumulation, but instead other by-products were formed. This minireview gives a clear outline of these research efforts and the outcome of them, including bioprocess level approaches and genetic approaches. Recently, the latter seems to have some promising results

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“…Many data in literature showed that E. coli grows in salt-based chemically defined media [19,39] as long as an organic carbon source is provided [22], as well as in rich complex organic media. Nevertheless, higher complexities in medium composition can lead to a lower reproducibility of the cell metabolic response and, therefore, lower the possibility of a well-controlled process [31].…”

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confidence: 99%

“…However, much work has to be done in order to establish the instant and profile addition of the growth-limiting carbon, often glucose [19,32] or glycerol [38].…”

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Application of a Fed-Batch Bioprocess for the Heterologous Production of hSCOMT in Escherichia coli

Passarinha

Bonifácio²,

Queiroz³

2009

J.Microbiol.Biotechnol

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In this paper, a fed-batch cultivation process in recombinant Escherichia coli BL21(DE3) bacteria, for the production of human soluble catechol-O-methyltransferase (hSCOMT), is presented. For the first time, a straightforward model is applied in a recombinant hSCOMT expression system and distinguishes an initial cell growth phase from a protein production phase upon induction. Specifically, the kinetic model predicts biomass, substrate, and product concentrations in the culture over time and was identified from a series of fed-batch experiments designed by testing several feed profiles. The main advantage of this model is that its parameters can be identified more reliably from distinct fed-batch strategies, such as glycerol pulses and exponential followed by constant substrate additions. Interestingly, with the limited amount of data available, the proposed model accomplishes satisfactorily the experimental results obtained for the three state variables, and no exhaustive process knowledge is required. The comparison of the measurement data obtained in a validation experiment with the model predictions showed the great extrapolation capability of the model presented, which could provide new complementary information for the COMT production system.

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Determination of the maximum specific uptake capacities for glucose and oxygen in glucose‐limited fed‐batch cultivations of Escherichia coli

Cited by 85 publications

References 33 publications

Oxygen Consumption Rates of Bacteria under Nutrient-Limited Conditions

Oxygen Consumption Rates of Bacteria under Nutrient-Limited Conditions

Minimizing acetate formation in E. coli fermentations

Application of a Fed-Batch Bioprocess for the Heterologous Production of hSCOMT in Escherichia coli

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