title: Towards scalable production of a collagen-like protein from Streptococcus pyogenes for biomedical applications.
abstract: BACKGROUND
  Collagen has proved valuable as biomedical materials for a range of clinical applications, particularly in wound healing. It is normally produced from animal sources, such as from bovines, but concerns have emerged over transmission of diseases. Recombinant collagens would be preferable, but are difficult to produce. Recently, studies have shown that 'collagens' from bacteria, including Streptococcus pyogenes, can be produced in the laboratory as recombinant products, and that these are biocompatible. In the present study we have established that examples of bacterial collagens can be produced in a bioreactor with high yields providing proof of manufacture of this important group of proteins.
  
  
  RESULTS
  Production trials in shake flask cultures gave low yields of recombinant product, < 1 g/L. Increased yields, of around 1 g/L, were obtained when the shake flask process was transferred to a stirred tank bioreactor, and the yield was further enhanced to around 10 g/L by implementation of a high cell density fed-batch process and the use of suitably formulated fully defined media. Similar yields were obtained with 2 different constructs, one containing an introduced heparin binding domain. The best yields, of up to 19 g/L were obtained using this high cell density strategy, with an extended 24 h production time.
  
  
  CONCLUSIONS
  These data have shown that recombinant bacterial collagen from S. pyogenes, can be produced in sufficient yield by a scalable microbial production process to give commercially acceptable yields for broad use in biomedical applications.
authors.affiliation: CSIRO Materials Science and Engineering, Bayview Avenue, Clayton, VIC 3168, Australia.
authors.forename: Yong Y
authors.initials: YY
authors.lastname: Peng
authors.forename: Linda
authors.initials: L
authors.lastname: Howell
authors.forename: Violet
authors.initials: V
authors.lastname: Stoichevska
authors.forename: Jerome A
authors.initials: JA
authors.lastname: Werkmeister
authors.forename: Geoff J
authors.initials: GJ
authors.lastname: Dumsday
authors.forename: John A M
authors.initials: JA
authors.lastname: Ramshaw
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001672:Biocompatible Materials / Q000378:metabolism; D019149:Bioreactors / Q000382:microbiology; D003094:Collagen / Q000235:genetics / Q000378:metabolism*; D003470:Culture Media / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013297:Streptococcus pyogenes / Q000378:metabolism*; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D001672:Biocompatible Materials; D003470:Culture Media; D011994:Recombinant Proteins; D003094:Collagen