Quantitation of heparosan with heparin lyase III and spectrophotometry

Haichan Huang; Yingying Zhao; Shencong Lv; Weihong Zhong; Fuming Zhang; Robert J. Linhardt

doi:10.1016/j.ab.2013.10.029

Quantitation of heparosan with heparin lyase III and spectrophotometry

Haichan Huang, Yingying Zhao, Shencong Lv, Weihong Zhong, Fuming Zhang, Robert J. Linhardt

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

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.

Original language	English
Pages (from-to)	46-48
Number of pages	3
Journal	Analytical Biochemistry
Volume	447
Issue number	1
DOIs	https://doi.org/10.1016/j.ab.2013.10.029
State	Published - 15 Feb 2014
Externally published	Yes

Keywords

Enzymatic method
Escherichia coli K5
Heparin lyase III
Heparosan
Quantitative method

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10.1016/j.ab.2013.10.029

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title = "Quantitation of heparosan with heparin lyase III and spectrophotometry",

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",

author = "Haichan Huang and Yingying Zhao and Shencong Lv and Weihong Zhong and Fuming Zhang and Linhardt, {Robert J.}",

note = "Funding Information: This study was financially supported by the Natural Science Foundation of China ( 21076195 ), the Science Technology Department of Zhejiang Province ( 2011C24007 ), CSC Scholarship, and the Specialized Research Fund for the Doctoral Program of Higher Education ( 20113317110002 ). The authors thank Jian Liu (University of North Carolina at Chapel Hill) for supplying the E. coli K5 and E. coli BL21 (carrying the plasmid containing the HepC gene) used in this study. ",

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AU - Huang, Haichan

AU - Zhao, Yingying

AU - Lv, Shencong

AU - Zhong, Weihong

AU - Zhang, Fuming

AU - Linhardt, Robert J.

N1 - Funding Information: This study was financially supported by the Natural Science Foundation of China ( 21076195 ), the Science Technology Department of Zhejiang Province ( 2011C24007 ), CSC Scholarship, and the Specialized Research Fund for the Doctoral Program of Higher Education ( 20113317110002 ). The authors thank Jian Liu (University of North Carolina at Chapel Hill) for supplying the E. coli K5 and E. coli BL21 (carrying the plasmid containing the HepC gene) used in this study.

PY - 2014/2/15

Y1 - 2014/2/15

N2 - 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.

AB - 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.

KW - Enzymatic method

KW - Escherichia coli K5

KW - Heparin lyase III

KW - Heparosan

KW - Quantitative method

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JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 1

ER -

Quantitation of heparosan with heparin lyase III and spectrophotometry

Abstract

Keywords

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