中文摘要：

腺苷二磷酸葡萄糖焦磷酸化酶（AGPase）活力传统上采用32P同位素标记反应底物来测定,但因测定条件的限制而极大地影响了其应用。该研究依据荧光素酶催化荧光素生成发光氧化荧光素的原理,在优化基本反应体系、确立反应体系ATP浓度和荧光强度线性关系等基础上,初步建立了以荧光发光反应测定AGPase活力的新方法,并运用新方法测定了含有不同glgC基因拷贝数菌株的AGPase活力。测定结果显示,不同菌株AGPase活力随glgC拷贝数不同存在显著差异,且其变化趋势与理论预期一致,即新方法可用于AGPase活力的体外测定,且具有更加安全、灵敏、简便和成本低的特点。

英文摘要：

ADP-glucose pyrophosphorylase（AGPase）catalyzes the reaction for formation of ADP-glucose,which is the substrate of starch and glycogen biosynthesis in plant and in bacteria.The reaction is the key step in starch and glycogen biosynthesis.Traditionally,AGPase activity is assayed through quantifying changes of 32P radio activity before or after the reaction.The assay was limited by poor availability of experimental condition.In this paper we described a new method to assay AGPase activity,which was established based on the reaction catalized by luciferase to generate oxiluciferin.AGPase activities in three E.coli strains,containing different copies of glgC were evaluated with the new method.The results showed that AGPase activities shifted along with copy number of glgC,which was identical with expected theoretically.The results indicated that new method is able to assay AGPase activity instead of the traditional approach.The new method is safer,more sensible,simpler and cheaper.