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Optimization of culture on the overproduction of TRAIL in high-cell-density culture by recombinant Escherichia coli

Abstract

Different nutrient-feeding cultures were carried out in producing recombinant protein of truncated tumor necrosis factor related apoptosis-inducing ligand (TRAIL) (114–281 amino acids of TRAIL) in Escherichia coli strain C600/pBV-TRAIL. The effects of preinduction specific growth rate, postinduction carbon source (glucose and glycerol), and feeding strategies were investigated. The higher preinduction specific growth rate (μ=0.22 h−1) contributed to the increase in the TRAIL production, at which TRAIL was accumulated in bacterial cells as 7.2% of total cellular protein, corresponding to 1.99 g l−1 in contrast with 5.1% (1.29 g l−1) at preinduction specific growth rate (μ=0.1 h−1) during high-cell-density culture. Glycerol was superior to glucose as the postinduction carbon source for TRAIL production. Under similar culture conditions, the final concentration of TRAIL was produced 1.59-fold more when glycerol was used as postinduction carbon source than when glucose was used. At the same time, the results showed that it is efficient to adopt the pH-stat feeding strategy at postinduction for the overproduction of TRAIL. The TRAIL production was increased up to 4.51 g l−1, approximately 16.1% of total cellular protein. The mechanisms behind the preinduction specific growth rate effect on the expression level may be ascribed to the leakage secretion of acetate.

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Acknowledgements

This work was supported by a grant from the Ministry of Science and Technology (Key Project of the National High Technology Research and Development Program of China (863 Program): Nos. 2002AA2Z345A and 2004AA2Z3801).

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Correspondence to Ya-Ling Shen or Dong-Zhi Wei.

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Luo, Q., Shen, Y., Wei, D. et al. Optimization of culture on the overproduction of TRAIL in high-cell-density culture by recombinant Escherichia coli . Appl Microbiol Biotechnol 71, 184–191 (2006). https://doi.org/10.1007/s00253-005-0131-6

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Keywords

  • Specific Growth Rate
  • Feeding Strategy
  • Acetate Concentration
  • Total Cellular Protein
  • Exponential Feeding