Department of Life Science, National Institute of Technology, Rourkela 769 008, Odisha, India.
Bioresour Technol. 2012 Nov;123:686-9. doi: 10.1016/j.biortech.2012.07.045. Epub 2012 Jul 23.
A high porosity carbon cloth with immobilized FAD was employed as working electrode in electrochemical NADH-regeneration procedure. Carbon cloth was oxidized with hot acids to create surface carboxyl group and then coupled by adenine amino group of FAD with carbodiimide in the presence of N-hydroxysulfosuccinimide. The bioelectrocatalytic NADH-regeneration was coupled to the conversion of achiral substrate pyruvate into chiral product l-lactate by l-lactate dehydrogenase (l-LDH) within the same reactor. The conversion was completed at 96h in bioreactor with FAD-modified carbon cloth, resulting in about 6mM of l-lactate from 10mM of pyruvate. While with bare carbon cloth, the yield at 120h was around 5mM. Immobilized FAD on the surface of carbon cloth electrode facilitated it to carry electrons from electrode to electron transfer enzymes; thereby NADH-regeneration was accelerated to drive the enzymatic reaction efficiently.
一种具有固定化 FAD 的高孔隙率碳布被用作电化学 NADH 再生过程中的工作电极。碳布先用热酸氧化，在 N-羟基琥珀酰亚胺存在的情况下，通过 FAD 的腺嘌呤氨基与碳二亚胺偶联，在碳布表面产生羧基基团。生物电化学 NADH 再生与同反应器内乳酸脱氢酶（l-LDH）将手性底物丙酮酸转化为手性产物 l-乳酸的反应偶联。在生物反应器中使用 FAD 修饰的碳布，在 96 小时内完成转化，从 10mM 丙酮酸中得到约 6mM 的 l-乳酸。而使用裸碳布，在 120 小时时的产量约为 5mM。固定在碳布电极表面的 FAD 有助于其将电子从电极传递给电子转移酶；从而加速 NADH 再生，有效地驱动酶反应。
