A novel all-in-one strategy for purification and immobilization of β-1,3-xylanase directly from cell lysate as active and recyclable nanobiocatalyst

Microbial Cell Factories

Yue Qiu¹  Xin Liu¹  Guangya Zhang¹  Zhongqi Ge¹  Yunmen Chu¹  Lixi Cai² 

[1] Department of Bioengineering and Biotechnology, Huaqiao University, 361021, Xiamen, Fujian, China;Department of Bioengineering and Biotechnology, Huaqiao University, 361021, Xiamen, Fujian, China;Faculty of Basic Medicine, Putian University, 351100, Putian, Fujian, China;

关键词: β-1,3-xylanase;    Enzyme immobilization;    Silica nanoparticles;    SpyCatcher;    SpyTag;   

DOI  :  10.1186/s12934-021-01530-5

来源: Springer

【 摘 要 】

BackgroundExploring a simple and versatile technique for direct immobilization of target enzymes from cell lysate without prior purification is urgently needed. Thus, a novel all-in-one strategy for purification and immobilization of β-1,3-xylanase was proposed, the target enzymes were covalently immobilized on silica nanoparticles via elastin-like polypeptides (ELPs)-based biomimetic silicification and SpyTag/SpyCatcher spontaneous reaction. Thus, the functional carriers that did not require the time-consuming surface modification step were quickly and efficiently prepared. These carriers could specifically immobilize the SpyTag-fused target enzymes from the cell lysate without pre-purification.ResultsThe ELPs-SpyCatcher hardly leaked from the carriers (0.5%), and the immobilization yield of enzyme was up to 96%. Immobilized enzyme retained 85.6% of the initial activity and showed 88.6% of the activity recovery. Compared with free ones, the immobilized β-1,3-xylanase showed improved thermal stability, elevated storage stability and good pH tolerance. It also retained more than 70.6% of initial activity after 12 reaction cycles, demonstrating its excellent reusability.ConclusionsThe results clearly highlighted the effectiveness of the novel enzyme immobilization method proposed here due to the improvement of overall performance of immobilized enzyme in respect to free form for the hydrolysis of macromolecular substrates. Thus, it may have great potential in the conversion of algae biomass as well as other related fields.

【 授权许可】

CC BY