[High density fed-batch culture of Escherichia coli DH5 alpha/pDH-B2m with DO feed-back control of nutrient feeding]

Sheng Wu Gong Cheng Xue Bao. 2002 Nov;18(6):718-23.
[Article in Chinese]

Abstract

Optimization of cultivation condition of recombinant E. coli DH5 alpha/pDH-B2m and the condition suitable for expression of recombinant mature peptide of human bone morphogenetic protein-2 was carried out in 500 mL shaking flasks and then transferred to NBS Bioflo IV, a 20 L DO feed-back fed-batch culture system, to obtain rhBMP-2. The results indicate that keeping dissolved oxygen at 40% and controlling nutrient feeding rate with DO feed back strategy can obtain theoretically 3.59 g recombinant mature peptide of hBMP-2 per liter of broth, the final cell density OD600 reaches 57(22.8 g dry cell weight/L), and the expression of rhBMP-2 amounts to 30% of the total protein in E. coli.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Proteins / biosynthesis
  • Bone Morphogenetic Proteins / genetics*
  • Culture Media
  • Escherichia coli / genetics*
  • Fermentation
  • Glycerol / pharmacology
  • Recombinant Proteins / biosynthesis*
  • Time Factors
  • Transforming Growth Factor beta*

Substances

  • BMP2 protein, human
  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Proteins
  • Culture Media
  • Recombinant Proteins
  • Transforming Growth Factor beta
  • Glycerol