title: Expanding the Catalytic Promiscuity of Heparinase III from Pedobacter heparinus.
abstract: Glycosaminoglycans (GAG) lyases are useful biocatalysts for the preparation of oligosaccharides, but their substrate spectra are limited to the same family. Thus, the degradation activity across families of GAG lyases is advantageous and desirable for various applications. In this study, residue Lys130 at the substrate entrance of monomeric heparinase III from Pedobacter heparinus ATCC 13125 was replaced by cysteine, and the resulting mutant K130C showed novel catalytic activity in degrading hyaluronic acid without affecting its native activity toward heparin and heparan sulfate. The broadened catalytic promiscuity by mutant K130C was the result of dimerization through a disulfide bond to expand the substrate binding pocket. This bifunctional enzyme is potentially valuable in the degradation of different types of GAGs.
authors.affiliation: State Key Laboratory of Natural Medicines and Laboratory of Chemical Biology, China Pharmaceutical University, 24 Tongjia St., Nanjing, Jiangsu Province, 210009, P. R. China.
authors.forename: Yayun
authors.initials: Y
authors.lastname: Gu
authors.affiliation: State Key Laboratory of Natural Medicines and Laboratory of Chemical Biology, China Pharmaceutical University, 24 Tongjia St., Nanjing, Jiangsu Province, 210009, P. R. China.
authors.forename: Meiling
authors.initials: M
authors.lastname: Lu
authors.affiliation: State Key Laboratory of Natural Medicines and Laboratory of Chemical Biology, China Pharmaceutical University, 24 Tongjia St., Nanjing, Jiangsu Province, 210009, P. R. China.
authors.forename: Zongqiang
authors.initials: Z
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Natural Medicines and Laboratory of Chemical Biology, China Pharmaceutical University, 24 Tongjia St., Nanjing, Jiangsu Province, 210009, P. R. China.
authors.forename: Xuri
authors.initials: X
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Natural Medicines and Laboratory of Chemical Biology, China Pharmaceutical University, 24 Tongjia St., Nanjing, Jiangsu Province, 210009, P. R. China.
authors.forename: Yijun
authors.identifier: http://orcid.org/0000-0002-4920-152X
authors.initials: Y
authors.lastname: Chen
mesh_terms: D055162:Biocatalysis; D019281:Dimerization; D004220:Disulfides / Q000737:chemistry; D006025:Glycosaminoglycans / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D060485:Pedobacter / Q000201:enzymology*; D011133:Polysaccharide-Lyases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity
chemical_list: D004220:Disulfides; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D011133:Polysaccharide-Lyases; C021349:heparitinsulfate lyase
keywords: Pedobacter heparinus; catalytic promiscuity; dimerization; disulfide linkage; heparinase III