title: Production of a low molecular weight heparin using recombinant glycuronidase [corrected].
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.
authors.affiliation: College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou 310032, China.
authors.forename: Guowei
authors.initials: G
authors.lastname: Su
authors.affiliation: College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou 310032, China.
authors.forename: Linhong
authors.initials: L
authors.lastname: Li
authors.affiliation: College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou 310032, China.
authors.forename: Haichan
authors.initials: H
authors.lastname: Huang
authors.affiliation: College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou 310032, China. Electronic address: whzhong@zjut.edu.cn.
authors.forename: Weihong
authors.initials: W
authors.lastname: Zhong
authors.affiliation: Department of Biological Engineering, Zhejiang Gongshang University, Hangzhou 310018, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Yu
authors.affiliation: Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Fuming
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Department of Biological Science, Center for Biotechnology and Interdisciplinary Studies, 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.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Linhardt
mesh_terms: D003001:Cloning, Molecular; D004187:Disaccharides / Q000378:metabolism; D005966:Glucuronidase / Q000378:metabolism*; D006495:Heparin, Low-Molecular-Weight / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism; D007218:Industrial Microbiology; D060485:Pedobacter / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D004187:Disaccharides; D005966:Glucuronidase; D006495:Heparin, Low-Molecular-Weight; D006497:Heparan Sulfate; D011994:Recombinant Proteins; C560363:heparosan
keywords: Depolymerization; Heparan sulfate; Heparin; Heparosan; Pedobacter heparinus; Δ4,5 unsaturated glycuronidase