Genetically engineered proteins with two active sites for enhanced biocatalysis and synergistic chemo- and biocatalysis
Genetically engineered proteins with two active sites for enhanced biocatalysis and synergistic chemo- and biocatalysis
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DOI：
10.1038/s41929-019-0394-4
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发表时间：
2020-03-01
期刊：
影响因子：
37.8
通讯作者：
Ferrer, Manuel
中科院分区：
文献类型：
--
作者：
Alonso, Sandra;Santiago, Gerard;Ferrer, Manuel
酶工程正在开辟新的化学领域。在这里，作者报道了被改造成含有两个生物活性位点的酶--还表明一个位点可以转化为金属络合物催化剂--并证明了这种双位点在一系列催化过程中的效用。酶工程不仅允许从头创造催化已知生物反应的活性位点，其速率接近扩散极限，而且还产生了进行新的自然反应的非生物场所。然而，将多个活性位点工程化到单个蛋白质支架中的催化优势尚未建立。在这里，我们报告了具有生物和/或非生物来源的两个活性位点的蛋白质，用于改善天然和非天然催化。该方法提高了含两个生物位点的酯酶的催化性能，如酶效率、底物范围、立体选择性和最佳温度窗口。然后，其中一个活性位点被变态为用于氧化和Friedel-Crafts烷基化反应的金属络合物化学催化位点，从而促进单个蛋白质中的协同化学和生物催化。乙酸1-萘酯转化为1，4-萘醌（转化率约为100%）。100%）和巴豆酸乙烯酯和苯转化为3-苯基丁酸（>= 83%; e. e. >99.9%）。
Enzyme engineering is opening up new chemistries. Here, the authors report enzymes engineered to contain two biological active sites - also showing that one site can be converted to a metal-complex catalyst - and demonstrate the utility of such dual sites in a range of catalytic processes.Enzyme engineering has allowed not only the de novo creation of active sites catalysing known biological reactions with rates close to diffusion limits, but also the generation of abiological sites performing new-to-nature reactions. However, the catalytic advantages of engineering multiple active sites into a single protein scaffold are yet to be established. Here, we report on proteins with two active sites of biological and/or abiological origin, for improved natural and non-natural catalysis. The approach increased the catalytic properties, such as enzyme efficiency, substrate scope, stereoselectivity and optimal temperature window, of an esterase containing two biological sites. Then, one of the active sites was metamorphosed into a metal-complex chemocatalytic site for oxidation and Friedel-Crafts alkylation reactions, facilitating synergistic chemo- and biocatalysis in a single protein. The transformations of 1-naphthyl acetate into 1,4-naphthoquinone (conversion approx. 100%) and vinyl crotonate and benzene into 3-phenylbutyric acid (>= 83%; e.e. >99.9%) were achieved in one pot with this artificial multifunctional metalloenzyme.