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Related Concept Videos

Fermentation01:29

Fermentation

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.
Fermentation is a type of metabolic process that occurs in the absence of oxygen, where organic molecules such as glucose are broken down to produce energy. During this process, the...
Microbial Fermentation01:23

Microbial Fermentation

Fermentation is a crucial anaerobic metabolic process that enables microbes to derive energy from sugar without relying on oxygen or an electron transport chain. This process is fundamental to various biological and industrial applications and is classified based on the metabolic products generated.Role of Pyruvate in FermentationPyruvate and its derivatives serve as key electron acceptors in fermentative pathways. The oxidation of NADH to regenerate NAD+ is essential for the continuation of...
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Bacterial Growth Curve

The bacterial growth curve is a fundamental concept in microbiology that describes the dynamics of bacterial population growth in a closed system with controlled environmental conditions, such as temperature and nutrient availability. This curve is divided into four distinct phases: lag, log (exponential), stationary, and death phases, each reflecting a unique stage of bacterial adaptation and growth. During the lag phase, bacteria acclimate to their surroundings by synthesizing essential...
Operon Model01:23

Operon Model

The operon model represents a fundamental mechanism of gene regulation in prokaryotes, enabling coordinated expression of genes involved in related metabolic or functional pathways. Operons consist of structural genes, a promoter, and an operator, with transcription regulated by repressors, activators, and small effector molecules.Structure and Function of OperonsAn operon is a cluster of structural genes transcribed together under the control of a single promoter. The promoter region...
Stringent Response in E. coli01:23

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Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...
Bioreactor Controls-II01:18

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In aerobic fermentations, oxygen is vital for microbial growth and metabolite production. Since air comprises only about 20% oxygen and the gas is poorly soluble in water—just 9 ppm at 20°C—supplying sufficient oxygen becomes a critical challenge, especially in high-demand processes like yeast growth or citric acid production. Even a fully saturated broth may offer only a few seconds of oxygen availability.To address this, sterile or scrubbed air is introduced into the fermentor via a sparger...

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Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae
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Metabolic oscillations in an E. coli fermentation.

D C Andersen1, J Swartz, T Ryll

  • 1Cell Culture & Fermentation Research & Development, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA. andersen@gene.com

Biotechnology and Bioengineering
|September 6, 2001
PubMed
Summary
This summary is machine-generated.

Recombinant E. coli fermentations showed reproducible oxygen uptake oscillations, slowing growth and delaying protein production. These metabolic oscillations were resolved by isoleucine feeding or genetic repair, indicating aberrant isoleucine biosynthesis regulation.

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Area of Science:

  • Microbial Physiology
  • Metabolic Engineering
  • Biotechnology

Background:

  • Recombinant Escherichia coli (E. coli) fermentations are crucial for producing valuable proteins.
  • Unexplained physiological behaviors can impede efficient recombinant protein production.

Purpose of the Study:

  • To investigate the cause of reproducible oxygen uptake oscillations observed during recombinant E. coli fermentations.
  • To identify the metabolic underpinnings of these oscillations and develop solutions.

Main Methods:

  • Controlled fermentation experiments in 10-L and 1,000-L fermentors.
  • Analysis of intracellular metabolite pools (amino acids, ATP, GTP).
  • Testing nutritional (isoleucine feeding) and genetic (ilvG repair) interventions.

Main Results:

  • Consistent oxygen uptake oscillations occurred across different scales and feed strategies.
  • Oscillations correlated with impaired protein synthesis and growth.
  • Metabolite analysis implicated aberrant isoleucine biosynthesis regulation and starvation events.
  • Isoleucine feeding and genetic repair of ilvG mutation eliminated oscillations.

Conclusions:

  • The observed oscillations stem from a metabolic defect in isoleucine biosynthesis regulation in recombinant E. coli.
  • These findings highlight the complex physiological challenges in large-scale recombinant microbial fermentations.
  • Solutions involve metabolic intervention or genetic correction of E. coli K-12 strains.