Lewis B, Tallman M, McGuinness E
Givaudan Fragrances Corp., Teaneck, NJ 07666, USA.
Analyst. 2001 Jun;126(6):855-60. doi: 10.1039/b008596k.
NADP(H)-specific Bakers yeast glucose 6-phosphate dehydrogenase (BYG6PDH) was paired, in turn, with each of three different source glutamate dehydrogenases (GDHs): NAD(P)-specific bovine liver (BLGDH), NADP-specific Candida utilis (CUGDH) and NADP-specific Proteus sp. (PSGDH) to constitute three enzyme cycling systems; (i) BYG6PDH/BLGDH; (ii) BYG6PDH/CUGDH; and (iii) BYG6PDH/PSGDH. When incorporated into an enzymatic cycling/amplification system for NAD kinase and run on a centrifugal fast analyzer (CFA), the microbial source enzyme CUGDH gave rise to a seven-fold greater amplification rate [21.5 x 10(3) cycles-1 (cph)] relative to that realized (3 x 10(3) cph) using the BYG6PDH/BLGDH cycling pair previously reported. Either of these cycling systems can be used as a flexible and general-purpose module for robotic amplification and data collection of NADP(H) linked enzymes as a user's requirements dictate. Although the BYG6PDH/PSGDH cycling pair produced a respectable cycling rate (14.4 x 10(3) cph), for reasons discussed the PSGDH enzyme was not considered a suitable replacement for BLGDH in an NADP(H) cycling system.
将烟酰胺腺嘌呤二核苷酸磷酸（还原型）[NADP(H)]特异性的面包酵母葡萄糖6-磷酸脱氢酶（BYG6PDH）依次与三种不同来源的谷氨酸脱氢酶（GDH）配对：烟酰胺腺嘌呤二核苷酸（磷酸）[NAD(P)]特异性的牛肝谷氨酸脱氢酶（BLGDH）、NADP特异性的产朊假丝酵母谷氨酸脱氢酶（CUGDH）和NADP特异性的变形杆菌属谷氨酸脱氢酶（PSGDH），以构成三个酶循环系统；（i）BYG6PDH/BLGDH；（ii）BYG6PDH/CUGDH；以及（iii）BYG6PDH/PSGDH。当将这些系统整合到用于NAD激酶的酶循环/放大系统中，并在离心式快速分析仪（CFA）上运行时，相对于先前报道的使用BYG6PDH/BLGDH循环对所实现的放大率[3×10³循环/小时（cph）]，微生物来源的酶CUGDH产生了高7倍的放大率[21.5×10³循环/小时（cph）]。根据用户需求，这些循环系统中的任何一个都可用作灵活的通用模块，用于NADP(H)连接酶的机器人放大和数据收集。尽管BYG6PDH/PSGDH循环对产生了可观的循环速率（14.4×10³ cph），但由于所讨论的原因，在NADP(H)循环系统中，PSGDH酶不被认为是BLGDH的合适替代品。
