苍山节杆菌CSAJ-16中性尿囊的克隆、异源表达及鉴定。

Polish journal of microbiology Pub Date : 2023-09-20 eCollection Date: 2023-09-01 DOI:10.33073/pjm-2023-027
Xin Yan, Wei Hu, Yun-Guo Zhu, Qing-Qing Liu, Shuai Wang, Hong-Yan Liu, Dan Zhu, Zhi-Hua Lv, Lin-Hua Li, Yi-Rui Yin
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引用次数: 0

摘要

尿酸酶(或尿酸氧化酶)是一种参与嘌呤代谢的关键酶,通常用于治疗痛风、高尿酸血症和肿瘤溶解综合征等疾病。本研究从云南苍山土壤样品中分离到一株产尿酸酶菌株(CSAJ-16)。该菌株被鉴定为节杆菌CSAJ-16。基于基因序列比对,扩增并异源表达了节杆菌CSAJ-16的尿酸酶基因(命名为aruox)。重组尿酸酶(ArUOX)约为32kDa。ArUOX的最适pH和温度分别为pH 7和20°C。在37°C下孵育100分钟或在pH 6.0-8.6下孵育24小时后,ArUOX保持在50%以上的相对活性。此外,金属离子如K+、Mg2+、Ca2+、Ba2+和Pb2+可以显著提高ArUOX的活性(>200%)。这些酶性质表明ArUOX在药物酶和尿酸检测试剂盒中具有潜在的应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Cloning, Heterologous Expression, and Characterization of a Neutral Uricase from Arthrobacter sp. CSAJ-16 in Cangshan Mountain.

Uricase (or Urate oxidase), a key enzyme involved in purine metabolism, is commonly used in treating conditions such as gout, hyperuricemia, and tumor lysis syndrome. In this study, a uricase-producing strain (named CSAJ-16) was isolated from the soil sample of Cangshan Mountain, Yunnan Province, China. This strain was identified as Arthrobacter sp. CSAJ-16. Based on the gene sequence alignment, the uricase gene (named aruox) of Arthrobacter sp. CSAJ-16 was amplified and heterologously expressed. The recombinant uricase (ArUOX) was about 32 kDa. The optimal pH and temperature of ArUOX were pH 7 and 20°C, respectively. The ArUOX remained above 50% relative activity after incubation at 37°C for 100 min or at pH 6.0-8.6 for 24 h. Moreover, metal ions such as K+, Mg2+, Ca2+, Ba2+ and Pb2+ can significantly enhance the activity of ArUOX (> 200%). These enzymatic properties indicate that ArUOX has potential applications in pharmaceutical enzymes and uric acid detection kits.

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