Cloning, sequence analysis, and expression in Escherichia coli of the gene encoding the Candida utilis urate oxidase (uricase).
Cloning, sequence analysis, and expression in Escherichia coli of the gene encoding the Candida utilis urate oxidase (uricase).
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DOI：
10.1093/oxfordjournals.jbchem.a021514
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发表时间：
1996-11
影响因子：
2.7
通讯作者：
Yasuji Koyama;Toshio Ichikawa;E. Nakano
中科院分区：
文献类型：
--
作者：
Yasuji Koyama;Toshio Ichikawa;E. Nakano
基于溴化氰裂解尿酸酶氨基酸序列，利用寡核苷酸探针从念珠菌中克隆出一个尿酸氧化酶基因。尿酸酶基因包含909个碱基对，编码的蛋白预计质量为34193 Da。念珠菌的尿酸酶与黄曲霉的尿酸酶相似（氨基酸序列匹配度为49%）。产自念珠菌的尿酸酶有4个半胱氨酸，其中一个胱氨酸参与酶活性。该酶在大肠杆菌细胞中以可溶性和功能性形式表达到约占细胞总蛋白20%的水平。
A urate oxidase (uricase) gene was cloned from Candida utilis with an oligonucleotide probe based on the amino acid sequence of cyanogen bromide-cleaved uricase. The uricase gene contains 909 base pairs and encodes a protein with a predicted mass of 34,193 Da. Candida uricase was similar (49% match in amino acid sequence) to the uricase from Aspergillus flavus. The uricase from Candida utilis has four cysteines and one of them, Cys168, participates in the enzyme activity. This enzyme was expressed to a level of about 20% of total cellular protein in an Escherichia coli cell as a soluble and functional form.