Format

Send to

Choose Destination
Anal Biochem. 2014 Feb 15;447:46-8. doi: 10.1016/j.ab.2013.10.029. Epub 2013 Oct 30.

Quantitation of heparosan with heparin lyase III and spectrophotometry.

Author information

1
College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou 310032, People's Republic of China.
2
College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou 310032, People's Republic of China. Electronic address: whzhong@zjut.edu.cn.
3
Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
4
Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Department of Biology, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Departments of Chemistry and Chemical Biology and Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.

Abstract

Heparosan is Escherichia coli K5 capsule polysaccharide, which is the key precursor for preparing bioengineered heparin. A rapid and effective quantitative method for detecting heparosan is important in the large-scale production of heparosan. Heparin lyase III (Hep III) effectively catalyzes the heparosan depolymerization, forming unsaturated disaccharides that are measurable using a spectrophotometer at 232 nm. We report a new method for the quantitative detection of heparosan with heparin lyase III and spectrophotometry that is safer and more specific than the traditional carbazole assay. In an optimized detection system, heparosan at a minimum concentration of 0.60 g/L in fermentation broth can be detected.

KEYWORDS:

Enzymatic method; Escherichia coli K5; Heparin lyase III; Heparosan; Quantitative method

PMID:
24184357
DOI:
10.1016/j.ab.2013.10.029
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center