为获得产低温葡萄糖氧化酶(glucose oxidase,GOD)的细菌,提高其产酶能力,研究GOD酶学性质,通过平板显色法从海泥样品中筛选低温GOD生产菌株,经形态特征、生理生化鉴定及分子生物学鉴定确定菌株分类。经诱变育种获得高产GOD菌株,并对所产GOD进行酶学性质研究。研究获得1株编号8-III的高产低温GOD细菌,鉴定为柠檬酸杆菌属(Citrobacters sp. 8-III),产GOD酶活力为0.75 U/mL。经诱变得到可稳定遗传的突变菌株8-III-a44,产GOD酶活力为1.77 U/mL,为原始菌株的2.36倍。最适pH为6.5,最适作用温度为15 ℃,在0~35 ℃有较高酶活力,为低温酶。并且添加K+、Ni2+对GOD活性有明显促进作用。该研究为其在海产品保鲜、饲料防腐方面的应用奠定基础。
This study aimed to obtain low-temperature glucose oxidase (GOD) producing bacteria, improve their enzyme-producing ability and study the properties of GOD. The low-temperature GOD producing strains were screened from sea sediments and identified. The strain with high yield of GOD was obtained by mutation breeding, and the properties of such GOD were studied. A strain 8-III with high yield of cold-active GOD was identified as Citrobacters sp. 8-III, and the activity of the GOD was 0.75 U/mL. Moreover, a mutant strain 8-III-a44 with high stability was obtained, the produced GOD activity was 1.77 U/mL, which was 2.36 times of that of strain 8-III. Furthermore, the optimal temperature and pH for the GOD produced by 8-III-a44 strain were 15 ℃ and 6.5, respectively. Besides, the enzyme was stable at 0-35 ?C, which indicated that it was a low-temperature enzyme. In addition, K+ and Ni2+ significantly promoted its activity. In conclusion, this study expands the source of GOD and lays a foundation for its applications in seafoods and feeds preservation.
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