| 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) |
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| Purification and characterization of the glucose oxidase from a marine strain Bacillus sp. CAMT22370 |
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NIE Qi-qian,MO Ri-jian,XU De-feng,SUN Li-jun,WANG Ya-ling,YE Ri-ying
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| 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. CAMT22370 glucose oxidase purification enzymatic characteristics |
