title: Cloning and characterization of a novel chondroitin sulfate/dermatan sulfate 4-O-endosulfatase from a marine bacterium.
abstract: Sulfatases are potentially useful tools for structure-function studies of glycosaminoglycans (GAGs). To date, various GAG exosulfatases have been identified in eukaryotes and prokaryotes. However, endosulfatases that act on GAGs have rarely been reported. Recently, a novel HA and CS lyase (HCLase) was identified for the first time from a marine bacterium (Han, W., Wang, W., Zhao, M., Sugahara, K., and Li, F. (2014) J. Biol. Chem. 289, 27886-27898). In this study, a putative sulfatase gene, closely linked to the hclase gene in the genome, was recombinantly expressed and characterized in detail. The recombinant protein showed a specific N-acetylgalactosamine-4-O-sulfatase activity that removes 4-O-sulfate from both disaccharides and polysaccharides of chondroitin sulfate (CS)/dermatan sulfate (DS), suggesting that this sulfatase represents a novel endosulfatase. The novel endosulfatase exhibited maximal reaction rate in a phosphate buffer (pH 8.0) at 30 °C and effectively removed 17-65% of 4-O-sulfates from various CS and DS and thus significantly inhibited the interactions of CS and DS with a positively supercharged fluorescent protein. Moreover, this endosulfatase significantly promoted the digestion of CS by HCLase, suggesting that it enhances the digestion of CS/DS by the bacterium. Therefore, this endosulfatase is a potential tool for use in CS/DS-related studies and applications.
authors: From the National Glycoengineering Research Center and State Key Laboratory of Microbial Technology, Shandong University, 27 South Shanda Road, Jinan 250100, China and.
authors: Wenshuang
authors: W
authors: Wang
authors: From the National Glycoengineering Research Center and State Key Laboratory of Microbial Technology, Shandong University, 27 South Shanda Road, Jinan 250100, China and.
authors: Wenjun
authors: W
authors: Han
authors: From the National Glycoengineering Research Center and State Key Laboratory of Microbial Technology, Shandong University, 27 South Shanda Road, Jinan 250100, China and.
authors: Xingya
authors: X
authors: Cai
authors: From the National Glycoengineering Research Center and State Key Laboratory of Microbial Technology, Shandong University, 27 South Shanda Road, Jinan 250100, China and.
authors: Xiaoyu
authors: X
authors: Zheng
authors: the Proteoglycan Signaling and Therapeutics Research Group, Faculty of Advanced Life Science, Hokkaido University Graduate School of Life Science, Sapporo 001-0021, Japan.
authors: Kazuyuki
authors: K
authors: Sugahara
authors: From the National Glycoengineering Research Center and State Key Laboratory of Microbial Technology, Shandong University, 27 South Shanda Road, Jinan 250100, China and fuchuanli@sdu.edu.cn.
authors: Fuchuan
authors: F
authors: Li
mesh_terms: D001419:Bacteria / Q000201:enzymology*; D002809:Chondroitin Sulfates / Q000378:metabolism*; D002810:Chondroitinases and Chondroitin Lyases / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D008386:Marine Biology
chemical_list: D002809:Chondroitin Sulfates; D002810:Chondroitinases and Chondroitin Lyases
keywords: Chondroitin Sulfate; Dermatan Sulfate; Endosulfatase; Glycosaminoglycan; Marine Bacterium; Oligosaccharide; Polysaccharide; Sulfated Polysaccharide
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