Engineering Thermostability in Artificial Metalloenzymes to Increase Catalytic Activity
Engineering Thermostability in Artificial Metalloenzymes to Increase Catalytic Activity
复制标题
DOI：
10.1021/acscatal.0c05413
复制
发表时间：
2021-03-08
期刊：
影响因子：
12.9
通讯作者：
Kamer, Paul C. J.
中科院分区：
文献类型：
--
作者：
Doble, Megan, V;Obrecht, Lorenz;Kamer, Paul C. J.
蛋白质工程已广泛用于改善酶的工业应用，并通过提高酶的热稳定性和溶剂耐受性来拓宽酶的工作条件。在这里，我们表明，蛋白质工程可用于增加人工金属酶的热稳定性。热稳定变体的人类固醇载体蛋白2L，修改绑定金属催化剂，通过合理的设计，使用结构数据和3DM数据库创建。对这些变体进行了测试，以确定增强蛋白质支架稳定性的突变，并观察到许多修饰的金属酶的熔化温度显著增加。承受更高反应温度的能力导致1-辛烯加氢裂化的活性增加，转化数提高了五倍以上，而对直链醛的选择性保持高达80%。
Protein engineering has shown widespread use in improving the industrial application of enzymes and broadening the conditions they are able to operate under by increasing their thermostability and solvent tolerance. Here, we show that protein engineering can be used to increase the thermostability of an artificial metalloenzyme. Thermostable variants of the human steroid carrier protein 2L, modified to bind a metal catalyst, were created by rational design using structural data and a 3DM database. These variants were tested to identify mutations that enhanced the stability of the protein scaffold, and a significant increase in melting temperature was observed with a number of modified metalloenzymes. The ability to withstand higher reaction temperatures resulted in an increased activity in the hydroformylation of 1-octene, with more than fivefold improvement in turnover number, whereas the selectivity for linear aldehyde remained high up to 80%.