a Laboratory of Applied Biotechnology, Department of Applied Biosciences , Ghent University , Ghent , Belgium.
Bioengineered. 2017 May 4;8(3):196-202. doi: 10.1080/21655979.2016.1222993. Epub 2016 Sep 20.
The evolution of natural modular proteins and domain swapping by protein engineers have shown the disruptive potential of non-homologous recombination to create proteins with novel functions or traits. Bacteriophage endolysins, cellulosomes and polyketide synthases are 3 examples of natural modular proteins with each module having a dedicated function. These modular architectures have been created by extensive duplication, shuffling of domains and insertion/deletion of new domains. Protein engineers mimic these natural processes in vitro to create chimeras with altered properties or novel functions by swapping modules between different parental genes. Most domain swapping efforts are realized with traditional restriction and ligation techniques, which become particularly restrictive when either a large number of variants, or variants of proteins with multiple domains have to be constructed. Recent advances in homology-independent shuffling techniques increasingly address this need, but to realize the full potential of the synthetic biology of modular proteins a complete homology-independent method for both rational and random shuffling of modules from an unlimited number of parental genes is still needed.
自然模块化蛋白质的进化和蛋白质工程师的结构域交换表明，非同源重组具有产生具有新功能或新特性的蛋白质的颠覆性潜力。噬菌体溶菌酶、纤维素酶和聚酮合酶是 3 种具有独特功能的天然模块化蛋白质的例子。这些模块化结构是通过广泛的重复、结构域的改组以及新结构域的插入/缺失而产生的。蛋白质工程师在体外模拟这些自然过程，通过在不同的亲本基因之间交换模块，创造具有改变的特性或新功能的嵌合体。大多数结构域交换工作都是通过传统的限制和连接技术来实现的，当需要构建大量变体或具有多个结构域的蛋白质变体时，这些技术会变得特别受限。最近同源无关改组技术的进步越来越多地满足了这一需求，但要充分发挥模块化蛋白质的合成生物学潜力，仍然需要一种完整的同源无关方法，用于从无限数量的亲本基因中进行模块的理性和随机改组。
