Effect and mechanism analysis of different linkers on efficient catalysis of subunit-fused nitrile hydratase
Effect and mechanism analysis of different linkers on efficient catalysis of subunit-fused nitrile hydratase
复制标题
不同连接体对亚基融合腈水合酶高效催化的影响及机制分析
DOI：
10.1016/j.ijbiomac.2021.03.103
复制
发表时间：
2021-03-31
影响因子：
8.2
通讯作者：
Zhou, Zhemin
中科院分区：
文献类型：
--
作者：
Guo, Junling;Cheng, Zhongyi;Zhou, Zhemin
Protein fusion using a linker plays an important role for protein evolution. However, designing suitable linkers for protein evolution is yet challenging and under-explored. To further clarify the regular pattern of suitable type of linker for fusion proteins, one nitrile hydratase (NHase) was used as a target protein and subunit fusion strategy was carried out to improve its efficient catalysis. Subunit-fused variants with three different types of linkers were constructed and characterized. All variants exhibited higher stability than that of the wild type. The longer the linker was, the higher stability NHase showed, however, too long linker affected NHase activity and expression. Among the three types of linkers, the alpha-helical linker seemed more suitable for NHase than flexible or rigid linkers. Though it is not clear how the linkers affecting the activity, structure analysis indicated that the stability improvement is dependent on the additional salt bridge, H-bond, and the subunit interface area increasing due to the linker insertion, among which the additional salt bridge and interface area were more important factors. The results described here may be useful for redesigning other enzymes through subunit fusion.(c) 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http:// creativecommons.org/licenses/by/4.0/).
