Production of recombinant protein G through high-density fermentation of engineered bacteria as well as purification

  • Authors:
    • Hu‑Cheng Zhang
    • Jun Yang
    • Guo‑Wei Yang
    • Xiao‑Jie Wang
    • Hai‑Tao Fan
  • View Affiliations

  • Published online on: April 27, 2015     https://doi.org/10.3892/mmr.2015.3688
  • Pages: 3132-3138
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Abstract

Recombinant Streptococcus Protein G (PG) is a cell wall protein, which, when combined with mammal immunoglobulin, is used in separating antibody technology. High‑density fermentation technologies using an engineered recombinant PG‑producing bacteria as well as PG separation and purification technologies have a direct impact on the availability and application of PG. Through primary and secondary seed cultivation, a recombinant E. coli strain was subjected to high‑density fermentation with controlled feed supplement concentration under stimulation with isopropyl β‑D‑1‑thiogalactopyranoside. The present study investigated the effect of factors including inoculum size, oxygen levels, pH and the cultivating method on the fermentation process, as well as the effect of the separation and purification technologies, including ultrasonication, nickel column affinity chromatography, Sephadex G‑25 gel filtration chromatography and diethylaminoethanol‑sepharose fast flow ion exchange chromatography on the yield and purity of PG. The efficiency of extraction was detected using SDS‑PAGE. High‑density fermentation yielded 80‑150 g/l of bacteria and 1 g PG was obtained from one liter broth. The present study delivered a highly efficient novel method via which PG can be obtained at a high concentration and a purity >95%.
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August-2015
Volume 12 Issue 2

Print ISSN: 1791-2997
Online ISSN:1791-3004

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Spandidos Publications style
Zhang HC, Yang J, Yang GW, Wang XJ and Fan HT: Production of recombinant protein G through high-density fermentation of engineered bacteria as well as purification. Mol Med Rep 12: 3132-3138, 2015
APA
Zhang, H., Yang, J., Yang, G., Wang, X., & Fan, H. (2015). Production of recombinant protein G through high-density fermentation of engineered bacteria as well as purification. Molecular Medicine Reports, 12, 3132-3138. https://doi.org/10.3892/mmr.2015.3688
MLA
Zhang, H., Yang, J., Yang, G., Wang, X., Fan, H."Production of recombinant protein G through high-density fermentation of engineered bacteria as well as purification". Molecular Medicine Reports 12.2 (2015): 3132-3138.
Chicago
Zhang, H., Yang, J., Yang, G., Wang, X., Fan, H."Production of recombinant protein G through high-density fermentation of engineered bacteria as well as purification". Molecular Medicine Reports 12, no. 2 (2015): 3132-3138. https://doi.org/10.3892/mmr.2015.3688