ARTP诱变及高效筛选高产葡萄糖氧化酶菌株

Screening Mutation of High Glucose Oxidase Activity Using Atmospheric and Room Temperature Plasmas

摘要:

以黑曲霉为出发菌株，采用常压室温等离子体（ARTP）注入技术对前培养后的出发菌株进行诱变，并结合改进后的邻联茴香胺高效平板显色圈进行初筛及摇瓶复筛。获得一株产葡萄糖氧化酶高产菌株，其产量由出发菌的45.6 U/mL提高到85.3 U/mL。通过对发酵关键条件碳酸钙和吐温-80添加量进行优化，结果显示酶活提高到125.6 U/mL，较原始菌株提高了175.4%。经6代传代培养，产葡萄糖氧化酶性能稳定。

Abstract:

Pre-cultured Aspergillus niger was treated using ARTP for the development of high glucose oxidase strain. Improved O-dianisidine color-developing circle method was used for preliminary screening and the mutant strains were re-screened by liquid state fermentation in flasks. Mutant strain T-6's enzyme activity increased from 45.6 U/mL to 85.3 U/mL and was selected for its high glucose oxidase-producing ability. Glucose oxidase activity reached 125.6 U/mL which was 175.4% higher than that of the original strain when the critical fermentation conditions，calcium carbonate and Tween-80 were optimized. The genetic expression of glucose oxidase was showed to be stable when the strain was cultured for 6 generations.