Ancestral sequence reconstruction and resurrection provides useful information for protein engineering, yet its alliance with directed evolution has been little explored. In this study, we have resurrected several ancestral nodes of fungal laccases dating back ∼500 to 250 million years. Unlike modern laccases, the resurrected Mesozoic laccases were readily secreted by yeast, with similar kinetic parameters, a broader stability, and distinct pH activity profiles. The resurrected Agaricomycetes laccase carried 136 ancestral mutations, a molecular testimony to its origin, and it was subjected to directed evolution in order to improve the rate of 1,3-cyclopentanedione oxidation, a β–diketone initiator commonly used in vinyl polymerization reactions.

中文翻译：

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祖先复活和真菌中生漆酶的定向进化。

祖先序列的重建和复活为蛋白质工程提供了有用的信息，但很少探索与定向进化的联盟。在这项研究中，我们已经复活了数个约500至2.5亿年的真菌漆酶祖先节。与现代漆酶不同，复生的中生漆酶很容易被酵母菌分泌，具有相似的动力学参数，更广泛的稳定性和独特的pH活性曲线。复活的姬松茸漆酶带有136个祖先突变，这是其起源的分子证明，并且对其进行定向进化以提高1,3-环戊二酮的氧化速率，这是乙烯基聚合反应中常用的β-二酮引发剂。