Production of a low molecular weight heparin production using recombinant glycuronidase

Guowei Su; Linhong Li; Haichan Huang; Weihong Zhong; Ping Yu; Fuming Zhang; Robert J. Linhardt

doi:10.1016/j.carbpol.2015.08.001

Production of a low molecular weight heparin production using recombinant glycuronidase

Guowei Su, Linhong Li, Haichan Huang, Weihong Zhong, Ping Yu, Fuming Zhang, Robert J. Linhardt

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

6 Scopus citations

Abstract

The Δ4,5 unsaturated uronate (4-deoxy-α-L-threo-hex-4-eno-pyranosyluronic acid) residue is produced through the depolymerization of heparin, heparosan, and heparan sulfate with heparin lyases. The recovery of unsaturated uronate containing products is necessary to prepare low molecular weight heparin (LMWH) from heparin or heparosan. In this study, the gene of Δ4,5 and Δ4,5^Δ20 unsaturated glycuronidase (EC# 3.2.1.56) from Pedobacter heparinus (formerly Flavobacterium heparinum) was cloned into pMAL-c2x plasmid. Its fusion protein with MBP was expressed in Escherichia coli TB1. After purification, Δ4,5 unsaturated glycuronidase was evaluated. The Δ4,5^Δ20 glycuronidase showed excellent activity on the unsaturated bonds of the different depolymerized products from Hep I, Hep II, and Hep III on heparin, heparosan, and heparan sulfate.

Original language	English
Pages (from-to)	151-157
Number of pages	7
Journal	Carbohydrate Polymers
Volume	134
DOIs	https://doi.org/10.1016/j.carbpol.2015.08.001
State	Published - 17 Aug 2015
Externally published	Yes

Keywords

Depolymerizationa
Heparan sulfate
Heparin
Heparosan
Pedobacter heparinus
Δ4,5 unsaturated glycuronidase

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10.1016/j.carbpol.2015.08.001

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title = "Production of a low molecular weight heparin production using recombinant glycuronidase",

abstract = "The Δ4,5 unsaturated uronate (4-deoxy-α-L-threo-hex-4-eno-pyranosyluronic acid) residue is produced through the depolymerization of heparin, heparosan, and heparan sulfate with heparin lyases. The recovery of unsaturated uronate containing products is necessary to prepare low molecular weight heparin (LMWH) from heparin or heparosan. In this study, the gene of Δ4,5 and Δ4,5Δ20 unsaturated glycuronidase (EC# 3.2.1.56) from Pedobacter heparinus (formerly Flavobacterium heparinum) was cloned into pMAL-c2x plasmid. Its fusion protein with MBP was expressed in Escherichia coli TB1. After purification, Δ4,5 unsaturated glycuronidase was evaluated. The Δ4,5Δ20 glycuronidase showed excellent activity on the unsaturated bonds of the different depolymerized products from Hep I, Hep II, and Hep III on heparin, heparosan, and heparan sulfate.",

keywords = "Depolymerizationa, Heparan sulfate, Heparin, Heparosan, Pedobacter heparinus, Δ4,5 unsaturated glycuronidase",

author = "Guowei Su and Linhong Li and Haichan Huang and Weihong Zhong and Ping Yu and Fuming Zhang and Linhardt, {Robert J.}",

year = "2015",

month = aug,

day = "17",

doi = "10.1016/j.carbpol.2015.08.001",

language = "English",

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AU - Su, Guowei

AU - Li, Linhong

AU - Huang, Haichan

AU - Zhong, Weihong

AU - Yu, Ping

AU - Zhang, Fuming

AU - Linhardt, Robert J.

PY - 2015/8/17

Y1 - 2015/8/17

N2 - The Δ4,5 unsaturated uronate (4-deoxy-α-L-threo-hex-4-eno-pyranosyluronic acid) residue is produced through the depolymerization of heparin, heparosan, and heparan sulfate with heparin lyases. The recovery of unsaturated uronate containing products is necessary to prepare low molecular weight heparin (LMWH) from heparin or heparosan. In this study, the gene of Δ4,5 and Δ4,5Δ20 unsaturated glycuronidase (EC# 3.2.1.56) from Pedobacter heparinus (formerly Flavobacterium heparinum) was cloned into pMAL-c2x plasmid. Its fusion protein with MBP was expressed in Escherichia coli TB1. After purification, Δ4,5 unsaturated glycuronidase was evaluated. The Δ4,5Δ20 glycuronidase showed excellent activity on the unsaturated bonds of the different depolymerized products from Hep I, Hep II, and Hep III on heparin, heparosan, and heparan sulfate.

AB - The Δ4,5 unsaturated uronate (4-deoxy-α-L-threo-hex-4-eno-pyranosyluronic acid) residue is produced through the depolymerization of heparin, heparosan, and heparan sulfate with heparin lyases. The recovery of unsaturated uronate containing products is necessary to prepare low molecular weight heparin (LMWH) from heparin or heparosan. In this study, the gene of Δ4,5 and Δ4,5Δ20 unsaturated glycuronidase (EC# 3.2.1.56) from Pedobacter heparinus (formerly Flavobacterium heparinum) was cloned into pMAL-c2x plasmid. Its fusion protein with MBP was expressed in Escherichia coli TB1. After purification, Δ4,5 unsaturated glycuronidase was evaluated. The Δ4,5Δ20 glycuronidase showed excellent activity on the unsaturated bonds of the different depolymerized products from Hep I, Hep II, and Hep III on heparin, heparosan, and heparan sulfate.

KW - Depolymerizationa

KW - Heparan sulfate

KW - Heparin

KW - Heparosan

KW - Pedobacter heparinus

KW - Δ4,5 unsaturated glycuronidase

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DO - 10.1016/j.carbpol.2015.08.001

M3 - Article

C2 - 26428111

AN - SCOPUS:84939237306

SN - 0144-8617

VL - 134

SP - 151

EP - 157

JO - Carbohydrate Polymers

JF - Carbohydrate Polymers

ER -

Production of a low molecular weight heparin production using recombinant glycuronidase

Abstract

Keywords

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