Highly Efficient Flavin–Adenine Dinucleotide Glucose Dehydrogenase Fused to a Minimal Cytochrome C Domain,Journal of the American Chemical Society

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Highly Efficient Flavin–Adenine Dinucleotide Glucose Dehydrogenase Fused to a Minimal Cytochrome C Domain
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2017-09-21 00:00:00 , DOI: 10.1021/jacs.7b07011
Itay Algov ₁ , Jennifer Grushka ₁ , Raz Zarivach _{1,

2} , Lital Alfonta ₁

Affiliation

Flavin–adenine dinucleotide (FAD) dependent glucose dehydrogenase (GDH) is a thermostable, oxygen insensitive redox enzyme used in bioelectrochemical applications. The FAD cofactor of the enzyme is buried within the proteinaceous matrix of the enzyme, which makes it almost unreachable for a direct communication with an electrode. In this study, FAD dependent glucose dehydrogenase was fused to a natural minimal cytochrome domain in its c-terminus to achieve direct electron transfer. We introduce a fusion enzyme that can communicate with an electrode directly, without the use of a mediator molecule. The new fusion enzyme, with its direct electron transfer abilities displays superior activity to that of the native enzyme, with a k_cat that is ca. 3 times higher than that of the native enzyme, a k_cat/K_M that is more than 3 times higher than that of GDH and 5 to 7 times higher catalytic currents with an onset potential of ca. (−) 0.15 V vs Ag/AgCl, affording higher glucose sensing selectivity. Taking these parameters into consideration, the fusion enzyme presented can serve as a good candidate for blood glucose monitoring and for other glucose based bioelectrochemical systems.

中文翻译：

高效的黄素-腺嘌呤二核苷酸葡萄糖脱氢酶融合到最小的细胞色素C结构域。

黄素-腺嘌呤二核苷酸（FAD）依赖性葡萄糖脱氢酶（GDH）是一种生物化学应用中使用的对热稳定，对氧不敏感的氧化还原酶。酶的FAD辅助因子埋在酶的蛋白质基质中，这使其几乎无法与电极直接通讯。在这项研究中，将FAD依赖性葡萄糖脱氢酶融合到其c末端的天然最小细胞色素结构域中，以实现直接电子转移。我们介绍一种无需使用介体分子即可直接与电极通讯的融合酶。这种新的融合酶具有直接的电子转移能力，其活性要优于天然酶，其中的k _cat为约。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。比天然酶高3倍k _cat / K _M比GDH高3倍以上，催化电流高5到7倍，起始电势约为。（-）相对于Ag / AgCl为0.15 V，提供了更高的葡萄糖感测选择性。考虑到这些参数，提出的融合酶可以作为血糖监测和其他基于葡萄糖的生物电化学系统的良好候选者。

更新日期：2017-09-21

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