中文摘要：

10-羟基-2-癸烯酸（10-HDA）是蜂王浆中的主要脂肪酸成分,具有抗菌、抗癌、延缓衰老等多种生理活性,但目前关于10-HDA生物合成的分子机制还不清楚。通过克隆蜜蜂NADPH-细胞色素P450还原酶（EC 1.6.2.4,NADPH-cytochrome P450 reductase,CPR）,在大肠杆菌中异源表达,并对其酶学特性进行分析。结果表明重组菌经IPTG诱导后表达蛋白的分子量与预期一致,为86.29 kDa,Ni-NTA亲和纯化后测得其比活性为77.33（EU of CPR）/μg。酶学性质分析结果表明蜜蜂CPR酶最适温度与pH分别为40℃和8.0,并对一些金属离子及有机溶剂具有不同程度的耐受性。其对底物细胞色素C的动力学参数Km和kcat分别为76μM和268/min。以上研究为探究CPR在10-HDA生物合成途径中的功能奠定理论基础。

英文摘要：

10-Hydroxy-2-decenoic acid（ 10-HDA）,a major fatty acid component from royal jelly,is known to have various biologic activities in antibacteria,antitumour and lifespan-extending. However,it is unclear about the underlying molecular mechanism of its biosynthesis. A NADPH-cytochrome P450 reductase（ EC 1. 6. 2. 4,NADPH-cytochrome P450 reductase,CPR） gene from Apis mellifera was cloned and expressed in Escherichia coli. Subsequently,the enzymatic properties of CPR were analyzed. Results showed that the target protein was successfully expressed in E. coli after induced by IPTG,and its molecular weight was 86. 29 kDa. The CPR protein was then purified by Ni-NTA protein affinity chromatography and its specific activity was 77. 33（ EU of CPR）/μg. The purified CPR protein displayed the maximum activity at pH 8. 0 and 40℃. Its activity was not significantly affected by several metal ions except Ni^2 ＋,Zn^2 ＋and Co^2 ＋which caused a strong inhibition. The Km and kcat,towards cytochrome C at 30℃,were 76μM and 268/min,respectively. It lay a foundation for researching the function of CPR in 10-HDA biosynthesis pathway.