High cell density cultivation of Escherichia coli K4 in a microfiltration bioreactor: a step towards improvement of chondroitin precursor production
Open Access
- 16 February 2011
- journal article
- Published by Springer Science and Business Media LLC in Microbial Cell Factories
- Vol. 10 (1), 10
- https://doi.org/10.1186/1475-2859-10-10
Abstract
The bacteria Escherichia coli K4 produces a capsular polysaccharide (K4 CPS) whose backbone is similar to the non sulphated chondroitin chain. The chondroitin sulphate is one of the major components of the extra-cellular matrix of the vertebrate connective tissues and a high value molecule, widely employed as active principle in the treatment of osteoarthritis. It is usually obtained by extraction from animal tissues, but the risk of virus contaminations, as well as the scarceness of raw material, makes this productive process unsafe and unable to satisfy the growing market demand. In previous studies a new biotechnological process to produce chondroitin from Escherichia coli K4 capsular polysaccharide was investigated and a 1.4 g·L-1 K4 CPS concentration was reached using fed-batch fermentation techniques. In this work, on the trail of these results, we exploited new fermentation strategies to further improve the capsular polysaccharide production. The inhibitory effect of acetate on the bacterial cells growth and K4 CPS production was studied in shake flask conditions, while a new approach, that combined the optimization of the feeding profiles, the improvement of aeration conditions and the use of a microfiltration bioreactor, was investigated in three different types of fermentation processes. High polysaccharide concentrations (4.73 ± 0.2 g·L-1), with corresponding average yields (0.13 ± 0.006 gK4 CPS·gcdw -1), were obtained; the increase of K4 CPS titre, compared to batch and fed-batch results, was of 16-fold and 3.3-fold respectively, while average yield was almost 3.5 and 1.4 fold higher. The increase of capsular polysaccharide titre confirmed the validity of the proposed fermentation strategy and opened the way to the use of the microfiltration bioreactor for the biotechnological production of chondroitin.Keywords
This publication has 24 references indexed in Scilit:
- High cell density cultivation and recombinant protein production with Escherichia coli in a rocking-motion-type bioreactorMicrobial Cell Factories, 2010
- High-performance liquid chromatography-mass spectrometry for mapping and sequencing glycosaminoglycan-derived oligosaccharidesNature Protocols, 2010
- High‐performance CE of Escherichia coli K4 cell surface polysaccharidesElectrophoresis, 2009
- Optimization of medium components for high-molecular-weight hyaluronic acid production by Streptococcus sp. ID9102 via a statistical approachJournal of Industrial Microbiology & Biotechnology, 2009
- Minimizing acetate formation in E. coli fermentationsJournal of Industrial Microbiology & Biotechnology, 2007
- Hyaluronic acid: a natural biopolymer with a broad range of biomedical and industrial applicationsBiotechnology Letters, 2006
- Exopolysaccharides production in Lactobacillus bulgaricus and Lactobacillus casei exploiting microfiltrationJournal of Industrial Microbiology & Biotechnology, 2006
- Enhanced production of human epidermal growth factor by a recombinant Escherichia coli integrated with in situ exchange of acetic acid by macroporous ion-exchange resinJournal of Bioscience and Bioengineering, 2005
- Growing E. coli to high cell density—A historical perspective on method developmentBiotechnology Advances, 2005
- Effective production of a thermostable α-glucosidase fromSulfolobus solfataricus inEscherichia coli exploiting a microfiltration bioreactorBiotechnology & Bioengineering, 2000