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首页> 外文期刊>Chemistry: A European journal >Fructose 1, 6-Bisphosphate Aldolase from Staphylococcus carnosus: Overexpression, Structure Prediction, Stereoselectivity, and Application in the Synthesis of Bicyclic Sugars
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Fructose 1, 6-Bisphosphate Aldolase from Staphylococcus carnosus: Overexpression, Structure Prediction, Stereoselectivity, and Application in the Synthesis of Bicyclic Sugars

机译：葡萄球菌的果糖1，6-二磷酸醛缩醛醛糖酶：过表达，结构预测，立体选择性及其在双环糖合成中的应用

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摘要

The gene for the fructose 1, 6-bisphosphate aldolase from Staphylococcus carnosus (FruA_(sca)) was subcloned for overexpression in Escherichia coli using the expression vector pKK223-3. An efficient, single-step purification by DEAE ion-exchange chromatography furnished the recombinant enzyme ready for synthetic applications. Sequence analysis indicated that FruA_(sca) shares the overall #alpha#/#beta#-barrel structure and most of the active site residues with the structurally well-defined FruA catalyst from rabbit muscle which signaled its functional equivalence for synthetic applications. A preparative study with generic aliphatic and hydroxylated aldehydes indeed confirmed a high level of stereoselectivity for both newly created asymmetric centers, and suggested a kinetic enantioselectivity for anionically charged 3-hydroxyaldehydes. In fact, the monomeric FruA_(sca) was found to tolerate even the presence of highly reactive glutardialdehyde derivatives, which otherwise rapidly denature the rabbit muscle enzyme, and to allow their stereoselective conversion to bicyclic sugars.
机译：使用表达载体pKK223-3，亚克隆来自葡萄球菌的果糖1、6-二磷酸醛缩酶的基因（FruA_（sca）），使其在大肠杆菌中过表达。通过DEAE离子交换色谱法进行的高效，一步纯化可为重组酶提供合成应用准备。序列分析表明，FruA_（sca）与来自兔肌肉的结构明确的FruA催化剂共享整体的#alpha＃/＃beta＃-桶状结构和大多数活性位点残基，这表明其功能等同于合成应用。使用通用的脂肪族和羟基化醛类进行的制备研究确实证实了两个新创建的不对称中心都具有较高的立体选择性，并提出了带阴离子电荷的3-羟基醛类的动力学对映选择性。实际上，发现单体FruA_（sca）甚至可以耐受高反应性戊二醛衍生物的存在，否则它们会迅速使兔肌肉酶变性，并使其立体选择性转化为双环糖。

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