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.
出处
《食品工业科技》
CAS
CSCD
北大核心
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)
关键词
RhaD
醛缩酶
一锅四酶法
磷酸二羟基丙酮
稀有糖
RhaD
aldolase
one- pot four-enzyme approach
dinydroxyacetone phosphate
rare sugar