Cloning and characterization of a hyaluronate lyase EsHyl8 from Escherichia sp. A99

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2024-07-10 DOI:10.1016/j.pep.2024.106551

Xiuli Cui , Zheng Fu , Hainan Wang , Wengong Yu , Feng Han

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引用次数: 0

Abstract

Hyaluronidase, an enzyme that degrades hyaluronic acid (HA), is utilized in clinical settings to facilitate drug diffusion, manage extravasation, and address injection-related complications linked to HA-based fillers. In this study, a novel hyaluronate lyase EsHyl8 was cloned, expressed, and characterized from Escherichia sp. A99 of human intestinal origin. This lyase belongs to polysaccharide lyase (PL) family 8, and showed specific activity towards HA. EsHyl8 exhibited optimal degradation at 40 °C and pH 6.0. EsHyl8 exhibited a high activity of 376.32 U/mg among hyaluronidases of human gut microorganisms. EsHyl8 was stable at 37 °C and remained about 70 % of activity after incubation at 37 °C for 24 h, demonstrating excellent thermostability. The activity of EsHyl8 was inhibited by Zn²⁺, Cu²⁺, Fe³⁺, and SDS. EsHyl8 was an endo-type enzyme whose end-product was unsaturated disaccharide. This study enhances our understanding of hyaluronidases from human gut microorganisms.

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来自 Escherichia sp. A99 的透明质酸裂解酶 EsHyl8 的克隆和表征

透明质酸酶是一种降解透明质酸（HA）的酶，在临床上用于促进药物扩散、控制外渗以及解决与基于透明质酸的填充剂相关的注射并发症。在这项研究中，克隆、表达并鉴定了一种新型透明质酸裂解酶 EsHyl8，该酶来自人类肠道源性大肠杆菌 A99。这种裂解酶属于多糖裂解酶（PL）家族 8，对 HA 具有特异性活性。EsHyl8 在 40°C 和 pH 值为 6.0 的条件下表现出最佳降解能力。在人类肠道微生物的透明质酸酶中，EsHyl8 表现出 376.32 U/mg 的高活性。EsHyl8 在 37 摄氏度下稳定，在 37 摄氏度下培养 24 小时后仍能保持约 70% 的活性，显示出极佳的热稳定性。Zn2+、Cu2+、Fe3+和SDS均可抑制EsHyl8的活性。EsHyl8 是一种内切型酶，其最终产物是不饱和二糖。这项研究加深了我们对人类肠道微生物透明质酸酶的了解。

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.