期刊
ACS APPLIED MATERIALS & INTERFACES
卷 11, 期 35, 页码 31878-31888出版社
AMER CHEMICAL SOC
DOI: 10.1021/acsami.9b09071
关键词
biomimetic mineralization; purification; enzyme immobilization; elastin-like polypeptides; biocatalyst
资金
- National Natural Science Foundation of China [21676124, 21878131]
- Opening Project of Henan Province Key Laboratory of Water Pollution Control and Rehabilitation Technology [CJSZ2018001, CJSZ2018009]
Inspired by natural biomineralization process, inorganic phosphates system has been selected as a candidate for the encapsulation of enzyme; however, during the long-term fabrication process, the loss of enzyme activity is unavoidable, and the biomimetic mineralization mechanism is still poorly understood. Meanwhile, the purification process plays a key role in the preparation of immobilized enzyme with high enzyme loading and activity, while the rapid, low-cost, and eco-friendly purification of biocatalyst from crude fermentation broth remains a critical challenge in biochemical engineering. Here, a binary tag composed of elastin-like polypeptide (ELP) and His-tag was presented for the first time to be fused with beta-glucosidase (Glu) to construct a recombinant Glu- linker-ELP-His (GLEH) with the aim of developing a fast synthesis strategy combining purification and immobilization processes for a biocatalyst with better stability and recyclability. The purification fold and activity recovery of GLEH reached 18.1 and 95.2%, respectively, once a single inverse transition cycling was conducted at 25 degrees C for 10 min. Then, efficient biomineralization of hybrid enzyme-Cu-3(PO4)(2) nanoflowers was realized in 15 min by the action of His-tag and ultrasonic-assisted reaction method. The activity recovery and relative activity reached the maximum at 90.3 and 111.0%, respectively. We demonstrate that the crystal growth process of a hybrid nanoflower involves obvious nucleation, self-assembly, and the Ostwald ripening process, and the enzyme GLEH acts as a binder to assemble Cu-3(PO4)(2) nanoflakes. The immobilized GLEH nanoflowers show outstanding operation stability and recyclability, and their catalytic efficiency is close to that of free Glu.
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