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Related Concept Videos

Fates of Pyruvate01:20

Fates of Pyruvate

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Pyruvate is the end product of glycolysis, where glucose is oxidized to pyruvate, simultaneously reducing NAD+ to NADH. Two molecules of ATP are also produced by substrate-level phosphorylation.
In aerobic organisms, pyruvate is metabolized via the citric acid cycle to produce reduced coenzymes NADH and FADH2. These coenzymes are then oxidized in the electron transport chain to produce ATP and, in the process, regenerate the NAD+ and FAD. As seen in some cell types and organisms, fermentation...
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Fermentation01:29

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Most eukaryotic organisms require oxygen to survive and function adequately. Such organisms produce large amounts of energy during aerobic respiration by metabolizing glucose and oxygen into carbon dioxide and water. However, most eukaryotes can generate some energy in the absence of oxygen by anaerobic metabolism.
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Updated: Jun 27, 2025

Medium Preparation for the Cultivation of Microorganisms under Strictly Anaerobic/Anoxic Conditions
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2-Stage microfermentations.

Shuai Li1, Zhixia Ye2, Eirik A Moreb2

  • 1Department of Chemistry, Duke University, Durham, NC, USA.

Metabolic Engineering Communications
|April 26, 2024
PubMed
Summary
This summary is machine-generated.

Engineered cell factories offer diverse product possibilities. New protocols standardize evaluating these microbial production strains using a two-stage fermentation process, simplifying strain engineering and scale-up.

Keywords:
2-StageDynamic controlE. coliHigh throughputMetabolic engineeringMicrobial strain evaluationMicrofermentationPhosphateProtein expression

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Area of Science:

  • Synthetic Biology
  • Metabolic Engineering
  • Biotechnology

Background:

  • Advances in DNA synthesis and genome editing enable rapid generation of numerous engineered cell factory variants.
  • Increased strain variant generation necessitates standardized, high-throughput evaluation methods.
  • Previous work established engineered *E. coli* strains and a 2-stage production process decoupling growth and production.

Purpose of the Study:

  • To detail the development of protocols for evaluating engineered *E. coli* strains in 2-stage microfermentations.
  • To provide standardized methods for assessing cell factory performance.
  • To adapt these protocols for diverse products, hosts, and triggers.

Main Methods:

  • Development of detailed protocols for 2-stage microfermentations.
  • Utilizing phosphate depletion as a trigger to halt growth and induce heterologous expression.
  • Focus on decoupling cellular growth (stage 1) from product formation in stationary phase (stage 2).

Main Results:

  • Established robust protocols for evaluating engineered *E. coli* strains in microfermentation.
  • Demonstrated adaptability of the 2-stage process for various protein and small molecule products.
  • Provided a framework for adapting methods to different cellular hosts and trigger mechanisms.

Conclusions:

  • The developed microfermentation protocols standardize and simplify the evaluation of engineered cell factories.
  • This approach facilitates higher-throughput assessment of strain variants.
  • The methodology is adaptable, supporting broader applications in synthetic biology and bioproduction.