Site-Specific and High-Loading Immobilization of Proteins by Using Cohesin–Dockerin and CBM–Cellulose Interactions

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• 作者：Mei Li ; Yi Yue ; Zi-Jian Zhang ; Zai-Yu Wang ; Tian-Wei Tan ; Li-Hai Fan
• 刊名：Bioconjugate Chemistry
• 出版年：2016
• 出版时间：July 20, 2016
• 年：2016
• 卷：27
• 期：7
• 页码：1579-1583
• 全文大小：395K
• 年卷期：0
• ISSN：1520-4812

文摘

Immobilization of enzymes enhances their properties for application in industrial processes as reusable and robust biocatalysts. Here, we developed a new immobilization method by mimicking the natural cellulosome system. A group of cohesin and carbohydrate-binding module (CBM)-containing scaffoldins were genetically engineered, and their length was controlled by cohesin number. To use green fluorescent protein (GFP) as an immobilization model, its C-terminus was fused with a dockerin domain. GFP was able to specifically bind to scaffoldin via cohesin–dockerin interaction, while the scaffoldin could attach to cellulose by CBM–cellulose interaction. Our results showed that this mild and convenient approach was able to achieve site-specific immobilization, and the maximum GFP loading capacity reached ∼0.508 μmol/g cellulose.