Email updates

Keep up to date with the latest news and content from Microbial Cell Factories and BioMed Central.

Open Access Research

Towards scalable production of a collagen-like protein from Streptococcus pyogenes for biomedical applications

Yong Y Peng, Linda Howell, Violet Stoichevska, Jerome A Werkmeister, Geoff J Dumsday and John A M Ramshaw*

Author Affiliations

CSIRO Materials Science and Engineering, Bayview Avenue, Clayton, VIC 3168, Australia

For all author emails, please log on.

Microbial Cell Factories 2012, 11:146  doi:10.1186/1475-2859-11-146

Published: 5 November 2012

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.

Keywords:
Bacterial collagen; Recombinant protein production; Bioreactor; High cell density; pCold vector; Defined medium; Fed-batch process