Evolvability of random polypeptides through functional selection within a small library.
Evolvability of random polypeptides through functional selection within a small library.
复制标题
通过小型文库中的功能选择实现随机多肽的进化。
DOI：
--
复制
发表时间：
2002
期刊：
影响因子：
--
通讯作者：
T. Yomo
中科院分区：
文献类型：
--
作者：
Asao Yamauchi;T. Nakashima;N. Tokuriki;M. Hosokawa;H. Nogami;S. Arioka;I. Urabe;T. Yomo
一个定向进化与噬菌体展示的随机多肽的约140个氨基酸残基，直到第六代下的选择的基础上的亲和力的过渡态类似物的酯酶反应。实验设计故意将观察限制在每代仅10个克隆。第一代由三个可溶性随机多肽和从先前构建的文库中任意选择的七个克隆组成。选择在一代中显示最高亲和力的克隆，并进行随机诱变以产生下一代的变体。即使在每代中仅10个任意选择的多肽内，也存在雅阁结合亲和力的能力的足够的变体。此外，所选多肽的结合能力显示出随世代逐渐连续增加。此外，纯化的选择的随机多肽表现出酯酶活性的逐渐但显著的增加。在小序列多样性内的功能开发的容易性意味着即使在随机多肽的小群体内也促使酶进化。
A directed evolution with phage-displayed random polypeptides of about 140 amino acid residues was followed until the sixth generation under a selection based on affinity to a transition state analog for an esterase reaction. The experimental design deliberately limits the observation to only 10 clones per generation. The first generation consists of three soluble random polypeptides and seven arbitrarily chosen clones from a previously constructed library. The clone showing the highest affinity in a generation was selected and subjected to random mutagenesis to generate variants for the next generation. Even within only 10 arbitrarily chosen polypeptides in each of the generations, there are enough variants in accord to capacity of binding affinity. In addition, the binding capacity of the selected polypeptides showed a gradual continuous increase over the generation. Furthermore, the purified selected random polypeptides exhibited a gradual but significant increase in esterase activity. The ease of the functional development within a small sequence variety implies that enzyme evolution is prompted even within a small population of random polypeptides.