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Related Concept Videos

The Significance of Membrane Transport01:44

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The transport of solutes across the cell membrane is essential for metabolic processes, like maintaining cell size and volume, generating the action potential, exchanging nutrients and gases, etc. Membrane transport can be either passive or active. It can be simple diffusion, facilitated, or mediated transport aided by transport proteins such as transporters and channels.
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Transporters are essential membrane transport proteins with functions related to cell nutrition, homeostasis, communication, etc. Approximately 7% of all genes in the human genome code for transporters or transporter-related proteins.
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Among the three main modes of HGT—transformation, conjugation, and transduction—transduction is unique in that it is mediated by bacteriophages, or bacterial viruses.Transduction occurs in two ways. Generalized transduction occurs during the lytic cycle of a bacteriophage infection. In this process, bacteriophages infect bacterial cells, replicate within them, and ultimately cause cell lysis, releasing newly assembled virions. Occasionally, random fragments of the bacterial genome...
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Enzymes like flippase, floppase, and scramblase transfer phospholipids from one layer to another in the membrane, thereby affecting membrane asymmetry.
Flippase
Eukaryotic flippases are type-IV P-type ATPases or P4-ATPases belonging to P-type ATPase family proteins that are membrane-bound pumps involved in the ATP-mediated transport of ions and molecules across the membrane. Flippases flip specific phospholipids from the outer to the inner leaflet of a membrane. All P4-ATPases have one...
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Carrier-mediated transport is a pivotal process in drug absorption, particularly for lipid-insoluble drugs, and encompasses facilitated diffusion and active transport. Facilitated diffusion allows drugs to move along their concentration gradient without energy expenditure, while active transport utilizes ATP to drive drug movement against this gradient.
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The plasma membrane, a critical structure in cellular biology, houses an array of transporters, or carrier proteins, interspersed within its lipid bilayer. These proteins play a crucial role in solute transport through facilitated diffusion, a form of passive diffusion that uses transporters to move the molecules across the membrane.
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Procedure for Adaptive Laboratory Evolution of Microorganisms Using a Chemostat
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Membrane transporter identification and modulation via adaptive laboratory evolution.

Mohammad S Radi1, Jesus E SalcedoSora2, Se Hyeuk Kim1

  • 1Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet 2800 Kgs, Lyngby, Denmark.

Metabolic Engineering
|May 22, 2022
PubMed
Summary
This summary is machine-generated.

Adaptive laboratory evolution identified novel membrane transporters and mutations for amino acid transport in Escherichia coli. This method efficiently discovers and engineers transporters for biotechnological applications.

Keywords:
Adaptive laboratory evolutionAmino acidsEscherichia coliMembrane transporters

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Membrane transport proteins are crucial for cellular functions and biotechnological applications.
  • A significant portion of membrane transporter families are poorly characterized, hindering their application.
  • Identifying and understanding transporter function is essential for strain engineering.

Purpose of the Study:

  • To identify novel membrane transporters and specific mutations that modulate their activity using adaptive laboratory evolution.
  • To characterize the functional roles of identified transporters in amino acid uptake.
  • To demonstrate the utility of identified transporters in microbial production systems.

Main Methods:

  • Adaptive laboratory evolution of Escherichia coli under increasing concentrations of L-histidine, L-phenylalanine, L-threonine, and L-methionine.
  • Whole genome sequencing to identify mutations in evolved strains.
  • Reverse engineering of mutations and functional validation using flow cytometry assays.
  • Overexpression of identified transporters in production strains.

Main Results:

  • Evolved strains exhibited tolerance to elevated amino acid concentrations.
  • Key mutations were identified in transporters leuE, yddG, yedA, brnQ, and rhtC.
  • Novel functional roles for yedA (methionine transport) and brnQ (threonine transport) were established.
  • Overexpression of yddG enhanced L-phenylalanine production.

Conclusions:

  • Adaptive laboratory evolution is a powerful tool for discovering and characterizing membrane transporters.
  • Specific mutations can effectively modulate transporter activity for improved substrate uptake or efflux.
  • The identified transporters and engineering strategies hold promise for biotechnological applications and strain development.