基于RhaD醛缩酶的“一锅四酶法”合成D-山梨糖和D-阿洛酮糖  

One-pot four-enzyme synthesis of D-sorbose and D-psicose based on aldolase RhaD

作　　者：高雅慧[1] 金则成 钱超[2] 李子杰[2] 中西秀树[2] 高晓冬[2] 

机构地区：[1]江南大学食品学院,江苏无锡214122 [2]江南大学生物工程学院糖化学与生物技术教育部重点实验室,江苏无锡214122

出　　处：《食品工业科技》2013年第21期210-214,共5页Science and Technology of Food Industry

基　　金：国家自然科学基金项目(21302069);教育部科学技术研究重大项目(313027);江南大学自主科研计划项目基金(JUSRP1003);江南大学工业生物技术教育部重点实验室开放课题(KLIB-KF201206);2013年江南大学"国家大学生创新创业训练计划"(201310295040)

摘　　要：利用L-1-磷酸鼠李树胶糖醛缩酶(RhaD)以较低成本同时合成稀有糖D-山梨糖和D-阿洛酮糖。为避免直接使用价格昂贵且不稳定的底物磷酸二羟基丙酮(DHAP),以便宜的底物DL-3-磷酸甘油作为前体,采用基于RhaD和磷酸酶(AP或YqaB)的"一锅四酶法"对D-山梨糖和D-阿洛酮糖的合成进行了优化。产物D-山梨糖和D-阿洛酮糖用TLC、HPLC进行检测,并分别采用硅胶、P-2凝胶、Ca2+离子交换树脂对产物进行纯化以及1H NMR对产物进行鉴定。结果表明"一锅四酶法"以及分离纯化方法可高效地应用于D-山梨糖和D-阿洛酮糖的实际生产。该方法的建立将为其他稀有糖及其衍生物的合成提供理论依据。Rare sugars D-sorbose and D-psicose were simultaneously synthesized with L- rhamnulose 1-phosphate aldolase（RhaD）at low cost.To avoid direct use Df dihydroxyacetone phosphate（DHAP）, a rather expensive and unstable substrate, a one- pot four- enzyme slrarategy based on RhaD and phosphatase （AP or YqaB） was adopted to optimize the synthesis of D-sorbose and D-psicose with a cheap substrate DL-glycerol 3-phosphate as the starting material.The products were firstly eletected by TLC and HPLC,then purified by silica gel,P-2 gel filtration and Ca2＋ exchange resin.Lastly,the products after purification were identified with t H NMR. The results demonstrated that one-pot four-enzyme strategy and purification method could be efficiently applied for the practical production of D-sorbose and D-psicose.Thi,,i approach could provide the theory basis for the synthesis of more other rare sugars and their derivatives.

关 键 词：RhaD 醛缩酶 一锅四酶法 磷酸二羟基丙酮 稀有糖 

分 类 号：TS201.2[轻工技术与工程—食品科学]