Blood Tolerant Laccase by Directed Evolution
Blood Tolerant Laccase by Directed Evolution
复制标题
DOI：
10.1016/j.chembiol.2013.01.001
复制
发表时间：
2013-02-21
影响因子：
--
通讯作者：
Alcalde, Miguel
中科院分区：
文献类型：
--
作者：
Mate, Diana M.;Gonzalez-Perez, David;Alcalde, Miguel
高氧化还原电位漆酶是一种功能强大的生物催化剂，在生物技术中有着广泛的应用。我们已经将白腐真菌中的耐热漆酶转化为耐血漆酶。适应这种漆酶的健身人体血液的特定组成（高于中性pH值，高氯化物浓度）需要几代的定向进化的替代复杂的血液介质。我们进化的漆酶在人血浆和血液中进行了测试，显示出催化活性，同时在T1铜位点保持高氧化还原电位。在T1位点的第二配位区域引入的突变改变了pH活性曲线，并大大降低了氯化物的抑制作用。漆酶可以适应在极端条件下发挥作用的概念证明为植入式纳米生物装置，化学合成和解毒提供了一系列机会。
High-redox potential laccases are powerful biocatalysts with a wide range of applications in biotechnology. We have converted a thermostable laccase from a white-rot fungus into a blood tolerant laccase. Adapting the fitness of this laccase to the specific composition of human blood (above neutral pH, high chloride concentration) required several generations of directed evolution in a surrogate complex blood medium. Our evolved laccase was tested in both human plasma and blood, displaying catalytic activity while retaining a high redox potential at the T1 copper site. Mutations introduced in the second coordination sphere of the T1 site shifted the pH activity profile and drastically reduced the inhibitory effect of chloride. This proof of concept that laccases can be adapted to function in extreme conditions opens an array of opportunities for implantable nanobiodevices, chemical syntheses, and detoxification.