School of Grain Science and Technology, Jiangsu University of Science and Technology, Zhenjiang 212100, China.
School of Agricultural and Food Sciences, Zhejiang Agriculture and Forestry University, Hangzhou 311300, China.
Molecules. 2021 Feb 23;26(4):1186. doi: 10.3390/molecules26041186.
spore display has become a field of increasing interest in the past two decades. To improve the efficiency of spore display, its directed modification was performed based on the cellulosome architecture by introducing onto them divergent cohesin (Coh) modules that can specifically bind to the target enzyme bearing the matching dockerins (Doc). In this study, five different pairs of cohesins and dockerins, selected from four cellulolytic microbes, were examined for their capabilities in displaying a tetrameric enzyme β-galactosidase from IAM11001 on the surface of WB600 spores. Immunofluorescence microscopy, western blotting, dot blotting, and enzyme assay was applied to confirm its surface expression. All the resultant five Coh-Doc based spore display can hydrolyze -nitrophenyl-β-D-galactopyranoside. Further, the optimized Coh-Doc based spore display exhibited the highest display efficiency. Overall, the results of current study may open new perspectives on the use of Coh-Doc interaction, which will find application in improving the efficiency of spore display.
在过去的二十年中，孢子展示已成为一个日益受到关注的领域。为了提高孢子展示的效率，基于细胞外酶复合物（cellulosome）的结构，通过引入可以特异性结合带有匹配 dockerin 的靶酶的分歧黏合(cohesin)模块，对其进行了定向修饰。在这项研究中，从四种纤维素分解微生物中选择了五对不同的黏合(cohesin)和 dockerin，用于考察它们在将来自 IAM11001 的四聚体酶β-半乳糖苷酶展示在 WB600 孢子表面的能力。应用免疫荧光显微镜、western blot、斑点印迹和酶测定来确认其表面表达。所有基于这五对黏合(cohesin)-dockerin 的孢子展示都能水解-硝基苯-β-D-半乳糖吡喃糖苷。此外，优化后的基于黏合(cohesin)-dockerin 的孢子展示表现出最高的展示效率。总体而言，本研究的结果可能为黏合(cohesin)-dockerin 相互作用的应用开辟新的视角，这将有助于提高孢子展示的效率。
