Department of Chemistry, University of Connecticut, Storrs, CT, USA.
Department of Material Science, University of Connecticut, Storrs, CT, USA.
Methods Mol Biol. 2022;2487:205-225. doi: 10.1007/978-1-0716-2269-8_14.
This chapter reports a single-step preparation of nanoarmored bi-enzyme systems assembled on 1-D and 2-D nanomaterials, with glucose oxidase and peroxidase enzymes as model systems for cascade bio-catalysis. This is a simple and facile method to both exfoliate the bulk 1D (carbon nanotubes, CNT) and 2D nanomaterials (α-Zirconium phosphate, α-ZrP) and bind the enzymes in a single step. Exfoliation of the bulk material enhances the accessible surface area of the materials for the enzyme binding, and it also boosts the diffusion of reagents from the bulk phase to the active sites of the bio-catalysts. For example, a mixture of horseradish peroxidase, glucose oxidase, and bovine serum albumin (BSA) were adsorbed on the surfaces of the α-ZrP nanoplates or carbon nanotubes (CNT) as the bulk materials are exfoliated simultaneously, in a one-step process. The resulting bio-catalysts were thoroughly characterized by powder X-ray diffraction, electron microscopy, biochemical and biophysical methods, while enzyme activity studies proved successful binding of enzymes with retention of activities or even enhancements in their specific activities. For example, GOx/HRP/BSA/CNT displayed 6 times the activity of a mixture of GOx/HRP/BSA, under otherwise identical conditions. Similarly, GOx/HRP/BSA/ZrP had 3.5 times the activity of the corresponding mixture of GOx/HRP/BSA, in the absence of the nanoplates. These robust nano-dispersions worked extraordinarily well as active bio-catalysts. These two kinds of fabricated biocatalyst dispersions are also highly stable.
本章报告了一种在一维和二维纳米材料上组装纳米装甲双酶体系的单步制备方法,以葡萄糖氧化酶和过氧化物酶酶作为级联生物催化的模型体系。这是一种简单易行的方法,可以同时剥离块状 1D(碳纳米管,CNT)和 2D 纳米材料(α-磷酸锆,α-ZrP),并在一步中结合酶。块状材料的剥离增强了材料对酶结合的可及表面积,并且还促进了试剂从体相扩散到生物催化剂的活性位点。例如,辣根过氧化物酶、葡萄糖氧化酶和牛血清白蛋白(BSA)的混合物被吸附在α-ZrP 纳米片或碳纳米管(CNT)的表面上,同时剥离块状材料,一步完成。通过粉末 X 射线衍射、电子显微镜、生化和生物物理方法对所得生物催化剂进行了彻底的表征,而酶活性研究证明了酶的成功结合,保留了其活性,甚至提高了其比活性。例如,在其他条件相同的情况下,GOx/HRP/BSA/CNT 的活性是 GOx/HRP/BSA 混合物的 6 倍。同样,在没有纳米片的情况下,GOx/HRP/BSA/ZrP 的活性是相应的 GOx/HRP/BSA 混合物的 3.5 倍。这些坚固的纳米分散体作为活性生物催化剂表现得非常出色。这两种制备的生物催化剂分散体也具有很高的稳定性。