Electrified Nanoconfined Biocatalysis with Rapid Cofactor Recycling
Electrified Nanoconfined Biocatalysis with Rapid Cofactor Recycling
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DOI：
10.1002/cctc.201901245
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发表时间：
2019-10-30
期刊：
影响因子：
4.5
通讯作者：
Armstrong, Fraser A.
中科院分区：
文献类型：
--
作者：
Megarity, Clare F.;Siritanaratkul, Bhavin;Armstrong, Fraser A.
在活细胞中，催化反应链（级联反应）的整体速率通过将酶纳米限制在微小的封闭体积中而大大增强：以这种方式呈现，相互依赖的催化剂高度集中，中间体和辅因子必须扩散的距离（活性位点到活性位点）可能很小。在利用这一原理的并行技术中，酶级联被供电、放大和真实的实时监测，因为它们协同工作，被纳米限制在导电金属氧化物电极的孔内。该技术，绰号电化学叶，通过利用由黄素酶，铁氧还蛋白NADP（+）还原酶（FNR）催化的烟酰胺腺嘌呤二核苷酸（NADP（H））再循环来模拟叶绿体生物合成。吸附在纳米孔的内壁上，FNR在电极和NADP（H）之间快速转移电子。这种活性与氧化还原酶偶联，氧化还原酶也被纳米限制在孔内，其使辅因子交联并选择性地合成所需产物或感测分析物。催化剂和辅因子的使用是非常有效的。该技术简单、廉价、适应性强，可扩展到微观水平（诊断）和宏观水平（有机合成）。尽管天然FNR对NADP（H）具有特异性，但该技术可以通过基因工程扩展以包括NAD（H）作为再循环辅因子。
In living cells, the overall rates of catalytic reaction chains (cascades) are massively enhanced by nanoconfinement of enzymes in tiny enclosed volumes: presented in such a way, interdependent catalysts are highly concentrated, and distances (active site-to-active site) across which intermediates and cofactors must diffuse, may be tiny. In a parallel technology exploiting this principle, enzyme cascades are powered, amplified, and monitored in real time as they work in concert, being nanoconfined within the pores of an electrically conductive metal oxide electrode. The technology, nicknamed the electrochemical leaf, mimics chloroplast biosynthesis by exploiting nicotinamide adenine dinucleotide (NADP(H)) recycling catalysed by the flavoenzyme, ferredoxin NADP(+) reductase (FNR). Adsorbed on the inner walls of the nanopores, FNR rapidly transfers electrons between the electrode and NADP(H). This activity is coupled to an oxidoreductase enzyme, also nanoconfined within the pores, which recycles the cofactor and selectively synthesises a desired product or senses an analyte. Use of catalyst and cofactor is very efficient. The technology is simple, inexpensive and adaptable, and scalable to both microscopic levels (diagnostics) and macroscopic levels (organic synthesis). Whereas native FNR is specific for NADP(H), the technology can be extended by genetic engineering to include NAD(H) as recycling cofactor.