聂绮倩¹, 莫日坚¹, 徐德峰¹, 孙力军¹, 王雅玲¹, 叶日英¹
广东海洋大学 食品科技学院, 广东省水产品加工与安全重点实验室, 广东省海洋食品工程技术研究中心,水产品深加工广东普通高等学校重点实验室

摘要:
前期筛选获得一株海洋源Bacillus sp. CAMT22370高产葡萄糖氧化酶菌株, 本研究进一步通过离子交换及分子筛层析使Bacillus sp. CAMT22370葡萄糖氧化酶纯化了2.12倍, 酶活得率32.37%, 比活力达到8.47 U/mg, 分子质量约为50 kDa。酶学特性研究表明, 此酶在低温下有较好的催化活性, 最适温度为30℃, 超过30℃时酶活迅速下降, 45℃保温2 h活力降至75%左右; 最适pH为6.0, 在5.0以下和7.0以上时酶活迅速降低, 9.0时损失35%; Ca²⁺和Mn²⁺有明显激活作用, Na⁺和Zn²⁺有轻微激活作用, K⁺和Mg²⁺对酶活无显著影响, 而Fe²⁺、Fe³⁺和Cu²⁺对酶活有明显抑制作用, 酶动力学参数Vmax及Km值分别为26.85(μmol/(L·min))和109.26(μmol/(L·min)), 以上酶学特性与现有酶制剂存在明显不同,研究结果为该葡萄糖氧化酶的开发利用奠定了初步基础。
关键词:Bacillus sp. CAMT22370葡萄糖氧化酶分离酶学特性
DOI：10.11759/hykx20171017001
分类号:
基金项目:广东省科技厅项目(2013B021100016, 2014B020205006); 农业部东海与远洋渔业资源开发利用重点实验室开放课题(2014K03); 广东海洋大学大学生创新课题(CXXL2014042)

Purification and characterization of the glucose oxidase from a marine strain Bacillus sp. CAMT22370
NIE Qi-qian,MO Ri-jian,XU De-feng,SUN Li-jun,WANG Ya-ling,YE Ri-ying
Abstract:
At previous study, a marine strain Bacillus sp. CAMT22370 with high activity of glucose oxidase was screened and in this research the purification and characterization of glucose oxidase were conducted with ion-exchange and gel filtration chromatography. The result showed that the glucose oxidase was purified 2.12 fold with the recovery yield of 32.37%, the specific enzyme activity of 8.47 U/mg and molecular weight of 50 kDa. Furthermore, the characterization of glucose oxidase from Bacillus sp. CAMT22370 indicated that desirable catalyzing ability was observed at low temperature with the optimal temperature ure at 30℃ and the activity sharply decreased when the temperature was above 30℃. Especially, 25% of the GOD activity lost when the enzyme was maintained at temperature of 45℃ for 2h. The optimal pH for catalyzing was found at 6.0 and the activity dropped rapidly when the pH value was below 5.0 or above 7.0 and the activity loss of 35% was observed at pH of 9.0. The metal ions of Ca² and Mn²⁺ obviously enhanced the enzyme activity and the evident inhibition on activity was observed by metal ions of Fe²⁺, Fe³⁺ and Cu²⁺. In addition, Na⁺ and Zn²⁺ slightly enhanced the activity while the ions of K⁺ and Mg²⁺ had no effect on the enzyme activity. The dynamics of Vmax and Km were 26.85μmol/(L·min) and 109.26 μmol/(L·min), respectively. This research demonstrated that there was evident difference in enzymatic characteristics between Bacillus sp. CAMT22370 and the reported data and set a preliminary base for developing and utilizing this glucose oxidase.
Key words:Bacillus sp. CAMT22370glucose oxidasepurificationenzymatic characteristics





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