从 Vibrio sp. H240 中克隆、表达和鉴定一种糖胺聚糖裂解酶。
Cloning, expression, and characterization of a glycosaminoglycan lyase from Vibrio sp. H240.
机构信息
School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China; Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, 1 Wenhai Road, Qingdao 266237, PR China; Provincial Key Laboratory of Glycoscience and Glycotechnology, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China; Key Laboratory of Marine Drugs, Ministry of Education, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China.
School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China; Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, 1 Wenhai Road, Qingdao 266237, PR China; Provincial Key Laboratory of Glycoscience and Glycotechnology, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China; Key Laboratory of Marine Drugs, Ministry of Education, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China.
出版信息
Enzyme Microb Technol. 2022 Mar;154:109952. doi: 10.1016/j.enzmictec.2021.109952. Epub 2021 Nov 24.
Glycosaminoglycan lyase is an effective tool for the functional studies of glycosaminoglycans and for the preparation of oligosaccharides. In this study, a new glycosaminoglycan lyase HCLaseV with a molecular weight of 90 kDa was cloned, expressed, and characterized from Vibrio sp. H240. The lyase belonged to the polysaccharide lyase (PL)- 8 family. HCLaseV showed enzyme activities toward chondroitin sulfate A, chondroitin sulfate B, chondroitin sulfate C, and hyaluronic acid. HCLaseV exhibited the highest activity against HA at pH 7.0 and 40 °C. HCLaseV was an endo-type enzyme whose degradation end-product was unsaturated disaccharides. Ca inhibited the activity of HCLaseV to a certain extent, which was different from most of the enzymes in the PL-8 family. Mutagenesis studies showed that the Ca inhibition might be related to the Asn residue. The sequence homology was evaluated by mutagenesis studies, and the catalytic residues in HCLaseV were presumed to be His, Trp, and Tyr. These characteristics are helpful for further basic research and application.
糖胺聚糖裂解酶是研究糖胺聚糖功能和寡糖制备的有效工具。本研究从弧菌属 H240 中克隆、表达并鉴定了一种分子量为 90 kDa 的新型糖胺聚糖裂解酶 HCLaseV。该裂解酶属于多糖裂解酶（PL）-8 家族。HCLaseV 对硫酸软骨素 A、硫酸软骨素 B、硫酸软骨素 C 和透明质酸均具有酶活性。HCLaseV 在 pH 7.0 和 40°C 时对 HA 的活性最高。HCLaseV 是一种内酶，其降解末端产物为不饱和二糖。Ca 对 HCLaseV 的活性有一定的抑制作用，这与 PL-8 家族中的大多数酶不同。突变研究表明，Ca 抑制可能与 Asn 残基有关。通过突变研究评估了序列同源性，推测 HCLaseV 的催化残基为 His、Trp 和 Tyr。这些特性有助于进一步的基础研究和应用。
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