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In vivo self-aggregation and efficient preparation of recombinant lichenase based on ferritin / 生物工程学报
Chinese Journal of Biotechnology ; (12): 1602-1611, 2022.
Article in Chinese | WPRIM | ID: wpr-927804
ABSTRACT
Enzyme separation, purification, immobilization, and catalytic performance improvement have been the research hotspots and frontiers as well as the challenges in the field of biocatalysis. Thus, the development of novel methods for enzyme purification, immobilization, and improvement of their catalytic performance and storage are of great significance. Herein, ferritin was fused with the lichenase gene to achieve the purpose. The results showed that the fused gene was highly expressed in the cells of host strains, and that the resulted fusion proteins could self-aggregate into carrier-free active immobilized enzymes in vivo. Through low-speed centrifugation, the purity of the enzymes was up to > 90%, and the activity recovery was 61.1%. The activity of the enzymes after storage for 608 h was higher than the initial activity. After being used for 10 cycles, it still maintained 50.0% of the original activity. The insoluble active lichenase aggregates could spontaneously dissolve back into the buffer and formed the soluble polymeric lichenases with the diameter of about 12 nm. The specific activity of them was 12.09 times that of the free lichenase, while the catalytic efficiency was 7.11 times and the half-life at 50 ℃ was improved 11.09 folds. The results prove that the ferritin can be a versatile tag to trigger target enzyme self-aggregation and oligomerization in vivo, which can simplify the preparation of the target enzymes, improve their catalysis performance, and facilitate their storage.
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Full text: Available Index: WPRIM (Western Pacific) Main subject: Enzymes, Immobilized / Biocatalysis / Ferritins / Glycoside Hydrolases Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2022 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Enzymes, Immobilized / Biocatalysis / Ferritins / Glycoside Hydrolases Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2022 Type: Article