Protein secretion biotechnology using Streptomyces lividans: large-scale production of functional trimeric tumor necrosis factor alpha (Journal article)

Pozidis, C./ Lammertyn, E./ Politou, A. S./ Anne, J./ Tsiftsoglou, A. S./ Sianidis, G./ Economou, A.


We evaluated the feasibility of large-scale production of biopharmaceuticals expressed as heterologous polypeptides from the Gram-positive bacterium Streptomyces lividans. As a model protein we used murine tumor necrosis factor alpha (mTNFalpha). mTNFalpha fused C-terminally to the secretory signal peptide of the subtilisin-inhibitor protein from Streptomyces venezuelae. Under appropriate fermentation conditions, significant amounts of mature mTNFalpha (80-120 mg/L) can be recovered from spent growth media. Efficient downstream processing allowing rapid purification of mTNFalpha from culture supernatants was developed. Importantly, the protein is recovered from the spent growth medium in its native trimeric state as judged by biophysical analysis. Further, mTNFalpha secreted by S. lividans is significantly more active in an in vitro apoptosis tissue culture assay than a corresponding polypeptide produced in Escherichia coli. This pilot study provides the first validation of S. lividans protein secretion as an alternative bioprocess for large-scale production of oligomeric proteins of potential therapeutic value.
Institution and School/Department of submitter: Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών Υγείας. Τμήμα Ιατρικής
Keywords: Bioreactors,Cell Culture Techniques/*methods,Culture Media/pharmacology,Feasibility Studies,Fermentation,Glucose/pharmacology,Pilot Projects,Polymers/metabolism,Quality Control,Recombinant Proteins/biosynthesis/secretion,Streptomyces/drug effects/*metabolism,Tumor Necrosis Factor-alpha/biosynthesis/chemistry/isolation &,purification/*secretion
URI: http://olympias.lib.uoi.gr/jspui/handle/123456789/18953
ISSN: 0006-3592
Link: http://www.ncbi.nlm.nih.gov/pubmed/11460252
http://onlinelibrary.wiley.com/store/10.1002/1097-0290(20010320)72:6<611::AID-BIT1026>3.0.CO;2-0/asset/26_ftp.pdf?v=1&t=h0dhin0k&s=79311a00ec2b3431d643e7afd06abf6432fe8bbd
Appears in Collections:Άρθρα σε επιστημονικά περιοδικά ( Ανοικτά)

Files in This Item:
File Description SizeFormat 
Pozidis-2001-Protein secretion bi.pdf354.74 kBAdobe PDFView/Open    Request a copy



 Please use this identifier to cite or link to this item:
http://olympias.lib.uoi.gr/jspui/handle/123456789/18953
  This item is a favorite for 0 people.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.