Double enzyme-catalyzed microreactors using capillary electrophoresis.
Double enzyme-catalyzed microreactors using capillary electrophoresis.
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使用毛细管电泳的双酶催化微反应器。
DOI：
10.1002/elps.1150190310
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Gomez,FA
中科院分区：
文献类型：
--
作者：
Zhao,DS;Gomez,FA
这项工作评估了双酶催化微反应器的概念，使用毛细管电泳（CE）。在电泳条件下在毛细管中迁移，将底物和两种酶分别注入缓冲液中并进行反应。随后通过CE测定反应程度和产物比率。使用两个模型系统证明了该概念：分别通过己糖激酶（HK，EC www.example.com）和腺苷三磷酸双磷酸酶（APY，EC www.example.com）将三磷酸腺苷（ATP）转化为二磷酸腺苷（ADP）和一磷酸腺苷（AMP）2.7.1.13.6.1.5，分别在葡萄糖转化为葡萄糖-6-磷酸和无机磷酸盐中，以及还原型烟酰胺腺嘌呤二核苷酸（NADH）的转化，分别通过乳酸脱氢酶（LDH，EC 1.1.1.27）和葡萄糖-6-磷酸脱氢酶（G6 PDH，EC 1.1.1.49）转化为烟酰胺腺嘌呤二核苷酸（NAD）并回到NADH，分别将丙酮酸转化为乳酸和将葡萄糖-6-磷酸（glc-6-P）转化为6-磷酸葡萄糖酸。这些程序说明了使用的毛细管作为一个双微反应器和电泳条件下的反应产物的定量容易。
This work evaluates the concept of a double enzyme‐catalyzed microreactor using capillary electrophoresis (CE). Migrating in a capillary under electrophoresis conditions, plugs of substrate and two enzymes are injected separately in buffer and allowed to react. Extent of reaction and product ratios were subsequently determined by CE. This concept is demonstrated using two model systems: the conversion of adenosine triphosphate (ATP) to adenosine diphosphate (ADP) and adenosine monophosphate (AMP) by hexokinase (HK, EC 2.7.1.1) and apyrase (APY, EC 3.6.1.5), respectively, in the conversion of glucose to glucose‐6‐phosphate and inorganic phosphate, respectively, and the conversion of nicotinamide adenine dinucleotide, reduced form (NADH), to nicotinamide adenine dinucleotide (NAD) and back to NADH by lactate dehydrogenase (LDH, EC 1.1.1.27) and glucose‐6‐phosphate dehydrogenase (G6PDH, EC 1.1.1.49), respectively, in the conversion of pyruvate to lactate and glucose‐6‐phosphate (glc‐6‐P) to 6‐phosphogluconate, respectively. These procedures illustrate the use of the capillary as a double microreactor and the ease of quantitation of reaction products under conditions of electrophoresis.