A Simple Technique for Direct Immobilization of Target Enzymes from Cell Lysates Based on the SpyTag/SpyCatcher Spontaneous Reaction

Li-Xi Cai; Yuan-Qing Lin; Yun-Meng Chu; Xiao-Pin Chen; Li-Xing Liu; Ming Zhang; Guang-Ya Zhang

doi:10.21769/BioProtoc.4282

A Simple Technique for Direct Immobilization of Target Enzymes from Cell Lysates Based on the SpyTag/SpyCatcher Spontaneous Reaction

Bio Protoc. 2022 Jan 5;12(1):e4282. doi: 10.21769/BioProtoc.4282.

Authors

Li-Xi Cai^{1

2}, Yuan-Qing Lin², Yun-Meng Chu², Xiao-Pin Chen¹, Li-Xing Liu¹, Ming Zhang¹, Guang-Ya Zhang²

Affiliations

¹ College of Basic Medicine, Putian University, Putian, PR China.
² Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen, PR China.

Abstract

Many of the current methods for enzyme purification and immobilization suffer from several drawbacks, such as requiring tedious multistep procedures or long preparation, and being environmentally unfriendly, due to the chemicals and conditions involved. Thus, a simple technique for direct purification and immobilization of target enzymes from cell lysates was proposed. The elastin-like polypeptides (ELPs)-SpyCatcher chimera could mediate the formation of silica carriers within seconds and the target enzymes were then covalently immobilized on silica carriers via SpyCatcher/SpyTag spontaneous reaction. These tailor-made carriers were easily prepared, with precisely controlled morphology and size, as well as none-consuming surface modification needed, which could specifically immobilize the SpyTag-fused target enzymes from the cell lysate without pre-purification.

Keywords: 3-xylanase; Elastin-like polypeptides; Enzyme immobilization; Silica nanoparticles; SpyCatcher; SpyTag; β-1.