玉米黄烷酮-3-羟化酶原核表达条件对香橙素生成量的影响  

Effect of prokaryotic expression conditions of Zea mays flavanone-3-hydroxylase on production of aromadendrin

作　　者：黄旭[1,2] 王振 刘娟[1,2] 肖妃垚 田苗苗[1,2] 郭佳婧[2] 单杨 HUANG Xu;WANG Zhen;LIU Juan;XIAO Feiyao;TIAN Miaomiao;GUO Jiajing;SHAN Yang(Longping Branch Graduate School,Hunan University,Changsha 410082,China;Institute of Agricultural Product Processing,Hunan Academy of Agricultural Sciences/Hunan Province Key Lab of Fruits&Vegetables Storage,Processing,Quality and Safety/Hunan Province International Joint Lab on Fruits&Vegetables Processing,Quality and Safety,Changsha 410125,China)

机构地区：[1]湖南大学隆平分院,湖南长沙410082 [2]湖南省农业科学院农产品加工研究所/果蔬贮藏加工与质量安全湖南省重点实验室/湖南省果蔬加工与质量安全国际科技创新合作基地,湖南长沙410125

出　　处：《食品与发酵工业》2022年第3期38-43,共6页Food and Fermentation Industries

基　　金：湖南省科技厅重点领域研发技术项目(2019NK2041);湖南省农业科技创新资金重点项目(2020CX47)。

摘　　要：香橙素是类黄酮代谢途径中重要的中间化合物,黄烷酮-3-羟化酶(flavanone-3-hydroxylase,F3H)能以柚皮素为底物催化C;位接羟基从而生成香橙素。为实现香橙素绿色发展,该研究利用微生物法,对来自玉米的Zmf3h构建大肠杆菌原核表达系统后诱导ZmF3H蛋白表达。通过发酵实验验证ZmF3H在胞内可溶性表达并成功催化生成香橙素。为提高香橙素产量,对蛋白表达阶段的菌体密度、异丙基-β-D-硫代半乳糖苷(isopropyl-β-D-thiogalactoside,IPTG)诱导剂浓度、诱导时间和诱导温度等条件进行优化实验。单因素试验结果表明各实验组分别在菌种密度OD;=0.8~1.0,IPTG终浓度为0.6 mmol/L,诱导温度在23℃,诱导8 h时可明显提高香橙素生成量,综合各单因素优化条件进行实验验证后发现香橙素生成量为(32.27±1.98)mg/L,与初始条件相比提高了60.39%。Aromadendrin is an important intermediate compound in flavonoid metabolism pathway.Flavanone-3-hydroxylase(F3 H)catalyzes hydroxyl grafting of naringin at C;site.Microbial method was used to achieve the green development of aromadendrin.ZmF3 H protein was expressed by Escherichia coli containing Zmf3h from Zea mays.Fermentation experiments confirmed that ZmF3 H was soluble in intracellular and successfully catalyzed the production of aromadendrin.The results showed that the production of aromadendrin was significantly increased when the final IPTG concentration was 0.6 mmol/L,the induction temperature was 23℃,the induction time was 8 h,and the cell density was between 0.8 to 1.0.The production of aromadendrin was(32.27±1.98)mg/L after optimization which was 60.39%higher than that of initial conditions.

关 键 词：香橙素 黄烷酮-3-羟化酶 原核表达 表达条件优化 

分 类 号：TS209[轻工技术与工程—食品科学]