High-level production of soluble tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL) in high-density cultivation of recombinant Escherichia coli using a combined feeding strategy

Biotechnol Lett. 2004 Jun;26(12):981-4. doi: 10.1023/b:bile.0000030043.84226.ac.

Abstract

Recombinant Escherichia coli strain C600/pBV-TRAIL (encoding for 114-281 amino acids of TRAIL's soluble fragment) produced a recombinant human tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL). Using a combined strategy of exponential feeding and pH-stat feeding, high concentrations of biomass (65 g dry wt l(-1)) and active soluble TRAIL (1.4 g l(-1)) were obtained within 30 h. The accumulation of acetate, which usually occurs during the process of high-density culture of Escherichia coli and especially in the induction stage of protein synthesis, was avoided.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis Regulatory Proteins
  • Bioreactors / microbiology*
  • Cell Culture Techniques / methods*
  • Cell Proliferation
  • Cloning, Molecular / methods
  • Escherichia coli / physiology*
  • Gene Expression Regulation, Bacterial / physiology
  • Glucose / metabolism
  • Humans
  • Membrane Glycoproteins / biosynthesis*
  • Membrane Glycoproteins / chemistry
  • Membrane Glycoproteins / genetics*
  • Membrane Glycoproteins / isolation & purification
  • Nitrogen / metabolism
  • Protein Engineering / methods*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • TNF-Related Apoptosis-Inducing Ligand
  • Tumor Necrosis Factor-alpha / biosynthesis*
  • Tumor Necrosis Factor-alpha / chemistry
  • Tumor Necrosis Factor-alpha / genetics*
  • Tumor Necrosis Factor-alpha / isolation & purification

Substances

  • Apoptosis Regulatory Proteins
  • Membrane Glycoproteins
  • Recombinant Proteins
  • TNF-Related Apoptosis-Inducing Ligand
  • TNFSF10 protein, human
  • Tumor Necrosis Factor-alpha
  • Glucose
  • Nitrogen