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

Microbial Cell Factories

Research

Yong Y Peng¹  Jerome A Werkmeister¹  Geoff J Dumsday¹  Linda Howell¹  Violet Stoichevska¹  John A M Ramshaw¹ 

[1] CSIRO Materials Science and Engineering, Bayview Avenue, VIC 3168, Clayton, Australia;

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

DOI  :  10.1186/1475-2859-11-146

 received in 2012-09-01, accepted in 2012-10-29,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundCollagen 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.ResultsProduction 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.ConclusionsThese 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.

【 授权许可】

Unknown   
© Peng et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.