使用毛细管电泳的柱上酶催化微反应：定量研究。

On-column enzyme-catalyzed microreactions using capillary electrophoresis: quantitative studies.

作者信息

Zhang Ying, Kaddis John, Silverio Catherine, Zurita Cecilia, Gomez Frank A

机构信息

Department of Chemistry and Biochemistry, California State University, Los Angeles 90032-8202, USA.

出版信息

J Capill Electrophor Microchip Technol. 2002;7(1-2):1-9.

PMID:11936761

Abstract

Capillary electrophoresis and on-column enzyme-catalyzed microreactor techniques were used to quantitate the reaction projects resulting from three model systems: i) the conversion of nicotinamide adenine dinucleotide (NAD) to nicotinamide adenine dinucleotide, reduced form (NADH) in the oxidation of glucose-6-phosphate (glc-6-p) to 6-phosphogluconate by glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49); ii) 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; iii) the conversion of fructose-1,6-bisphosphate to dihydroxyacetone phosphate and glyceraldehyde-3-phosphate by fructose-biphosphate aldolase (ALD, EC 4.1.2.13). Single and double microreactor techniques employing direct or indirect detection were used to follow the conversion of substrate to product(s). In addition, electrophoresis conditions including voltage, enzyme concentration, and mixing time of the reaction, were correlated to product distribution profiles.

摘要

采用毛细管电泳和柱上酶催化微反应器技术对三个模型系统产生的反应产物进行定量分析

i）在葡萄糖-6-磷酸脱氢酶（G6PDH，EC 1.1.1.49）催化葡萄糖-6-磷酸（glc-6-p）氧化为6-磷酸葡萄糖酸的过程中，烟酰胺腺嘌呤二核苷酸（NAD）转化为还原型烟酰胺腺嘌呤二核苷酸（NADH）；ii）在葡萄糖分别转化为葡萄糖-6-磷酸和无机磷酸的过程中，己糖激酶（HK，EC 2.7.1.1）和腺苷三磷酸双磷酸酶（APY，EC 3.6.1.5）分别将三磷酸腺苷（ATP）转化为二磷酸腺苷（ADP）和一磷酸腺苷（AMP）；iii）果糖-1,6-二磷酸醛缩酶（ALD，EC 4.1.2.13）将果糖-1,6-二磷酸转化为二羟基丙酮磷酸和3-磷酸甘油醛。采用直接或间接检测的单微反应器和双微反应器技术跟踪底物向产物的转化。此外，将电泳条件（包括电压、酶浓度和反应混合时间）与产物分布图谱相关联。