首页|基于RhaD醛缩酶的“一锅四酶法”合成D-山梨糖和D-阿洛酮糖

基于RhaD醛缩酶的“一锅四酶法”合成D-山梨糖和D-阿洛酮糖

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  • 国家科技期刊平台
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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.
One-pot four-enzyme synthesis of D-sorbose and D-psicose based on aldolase RhaD
Rare sugars D-sorbose and D-psicose were simultaneously synthesized with L-rhamnulose 1-phosphate aldolase(RhaD)at low cost.To avoid direct use of dihydroxyacetone phosphate (DHAP),a rather expensive and unstable substrate,a one-pot four-enzyme strategy 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 detected 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 1H 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.This approach could provide the theory basis for the synthesis of more other rare sugars and their derivatives.

RhaDaldolaseone-pot four-enzyme approachdihydroxyacetone phosphaterare sugar

高雅慧、金则成、钱超、李子杰、中西秀树、高晓冬

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江南大学食品学院,江苏无锡214122

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

RhaD 醛缩酶 一锅四酶法 磷酸二羟基丙酮 稀有糖

国家自然科学基金教育部科学技术研究重大项目江南大学自主科研计划项目江南大学工业生物技术教育部重点实验室开放课题2013年江南大学"国家大学生创新创业训练计划"

21302069313027JUSRP1003KLIB-KF201206201310295040

2013

食品工业科技
北京一轻研究院

食品工业科技

CSTPCDCSCD北大核心
影响因子：0.842
ISSN：1002-0306
年,卷(期)：2013.34(21)
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