海洋高产尿酸氧化酶菌株筛选鉴定及酶学性质研究

doi:10.13560/j.cnki.biotech.bull.1985.2018-1057

摘要/Abstract

摘要： 尿酸氧化酶广泛用于常规临床分析和临床药物,在医学治疗和诊断中的重要性日益增加。为得到高产尿酸氧化酶的优良菌株,以大连黄海海泥、海水为材料,依次分别采用透明圈法、酶偶联分光光度法进行初筛和复筛,获一株高产尿酸氧化酶菌株Z7,根据菌株Z7的16S rDNA基因序列和形态学、生理生化特征,该菌株被鉴定为苛求芽孢杆菌（Bacillus fastidiosus）。酶学性质研究表明：尿酸氧化酶蛋白的分子量约为33.1 kD;最适作用温度是25℃,酶活高达673.7 U/mg;最适作用pH为8.0,在 pH 8-9表现出较强的稳定性;Fe³⁺、Ca²⁺对尿酸氧化酶具有激活作用。Ag⁺、Hg²⁺对该酶抑制性较强,使其几乎丧失活性。该菌株产酶活性及稳定性良好,可为尿酸氧化酶的工业化生产奠定理论基础,并具有潜在开发价值。

关键词: 苛求芽孢杆菌, 鉴定, 尿酸氧化酶, 分离纯化, 酶学性质

Abstract: Uricase is used in routine clinical analysis and clinical medicine widely. Its importance in medical treatment and diagnosis is increasing. A high uricase-producting strain Z7 was screened from the Dalian Huanghai Sea mud and seawater by transparent circle method and enzyme coupling spectrophotometry,and identified as Bacillus fastidiosus by 16S rDNA phylogenetic analysis,morphology and physiological and biochemical characteristics. Results showed that the molecular weight of the protein was about 33 kD. The optimum temperature for uricase was 25℃,and specific activity was 673.7 U/mg . The optimum pH for uricase was 8.0,and it was stable in the pH range of 8.0 to 9.0. Its relative enzyme activity could still maintain more than 90% after 1 h’s incubation. The activation of Fe³⁺and Ca²⁺ on uricase was stimulative effects,while Ag⁺,Hg²⁺ showed strong inhibitory effects. Owing to the excellent chill enzyme activity and stability,Z7 can lay a theoretical foundation for the industrialized production of uricase and has potential development value.

Key words: Bacillus fastidious, identification, uricase, isolation and purification, enzymatic properties

张庆芳, 逄飞, 于爽, 肖景惠, 窦少华, 迟乃玉. 海洋高产尿酸氧化酶菌株筛选鉴定及酶学性质研究[J]. 生物技术通报, 2019, 35(7): 61-69.

ZHANG Qing-fang, PANG Fei, YU Shuang, XIAO Jing-hui, DOU Shao-hua, CHI Nai-yu. Screening and Identification of High Uricase-producting Strain from Marine and the Enzymatic Properties[J]. Biotechnology Bulletin, 2019, 35(7): 61-69.

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