诊断用耐热尿酸氧化酶菌株的筛选及条件优化  

Screening of the diagnostic and thermostable urate oxidase-producing strain and optimization of the fermentation conditions

作　　者：张玉然[1] 纪楠楠[1] 

机构地区：[1]济宁医学院生物科学学院,山东日照276826

出　　处：《中国生化药物杂志》2016年第9期154-157,共4页Chinese Journal of Biochemical Pharmaceutics

基　　金：山东省医药卫生科技发展计划项目(2015WS0416);济宁医学院博士科研启动基金项目(JY2015BS12);国家级大学生创新创业训练计划项目(201510443008)

摘　　要：目的分离筛选及鉴定产耐热尿酸氧化酶的菌株,优化发酵条件并研究酶学性质。方法利用透明圈法从高温酒曲中筛选产耐热尿酸氧化酶的菌株,扩增菌株16S rDNA序列并构建系统发育进化树进行鉴定,优化菌株产尿酸氧化酶的发酵条件并研究该酶的酶学性质。结果筛选获得一株产耐热尿酸氧化酶的菌株,经分子鉴定命名为Bacillus subtilis ZX-6。于优化后的最佳发酵产酶条件下(30℃、初始p H 8.1、4 m L/L玉米浆)尿酸氧化酶水平达135.9 U/L,比优化前提高了133.7%。该酶最适反应温度及p H分别为45℃和7.6,于37℃保温48 h后,其剩余酶活仍为17.2%,具备良好的热稳定性。结论产耐热尿酸氧化酶菌株筛选及发酵条件优化的成功为进一步的研究奠定了良好的基础。Objective To screen a thermostable urate oxidase-producing strain,optimize the fermentation conditions and study the enzymatic properties. Methods A urate oxidase-producing strain was screened from high temperature starter based on transparent circle method. Its 16 S r DNA sequence was then amplified and analyzed. Meanwhile,the phylogenetic trees were built. Optimization of the fermentation conditions from this strain was carried out. The enzymatic properties of urate oxidase were studied. Results A urate oxidase-producing strain,named Bacillus subtilis ZX-6 by molecular identification,was obtained. The production of urate oxidase under the optimized conditons（135. 9 U/L） was 133. 7% higher than before. The optimum reaction temperature and p H were 45 ℃ and 7. 6 respectively. The residual activity of urate oxidase at 37 ℃ for 48 h was still 17. 2%. Conclusion The successful screening of a thermostable urate oxidase-producing strain and optimization of the fermentation conditions will lay a foundation for the further research.

关 键 词：尿酸氧化酶 BACILLUS SUBTILIS ZX-6 筛选 鉴定 条件优化 酶学性质 

分 类 号：Q554[生物学—生物化学]