Huckel M, Wirth H J, Hearn M T
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
J Biochem Biophys Methods. 1996 Feb 5;31(3-4):165-79. doi: 10.1016/0165-022x(95)00035-p.
Four different proteases (trypsin, chymotrypsin, papain and pepsin) were covalently attached to the surface of a new type of porous zirconia, as well as a conventional porous silica, activated with 3-isothiocyanatopropyltriethoxy silane (NCS-silane). The immobilization efficiency onto the porous zirconia material was evaluated in terms of the amount of enzyme attached to the particles and from the biological activity remaining after the immobilization step. The results were compared with the corresponding experiments with a porous silica of similar surface area/g support material. In addition, the storage stability of the modified zirconia and silica biocatalysts were evaluated. These results indicated that specific immobilized enzyme biocatalysts can be achieved with this new zirconia support material which exhibits different properties to those observed with the more conventional silica-based materials. Moreover, the results with the enzyme-zirconia biocatalysts also indicate different characteristics when compared with data for the same enzymes immobilized under similar buffer conditions to organic support materials as previously described by various other investigators. The advantages of zirconia-based immobilized enzyme biocatalysts in terms of their density and chemical robustness are also described relative to other alternative support materials currently in use.
四种不同的蛋白酶(胰蛋白酶、胰凝乳蛋白酶、木瓜蛋白酶和胃蛋白酶)被共价连接到一种新型多孔氧化锆的表面,以及用3-异硫氰酸丙基三乙氧基硅烷(NCS-硅烷)活化的传统多孔二氧化硅表面。通过附着在颗粒上的酶量以及固定化步骤后剩余的生物活性来评估固定在多孔氧化锆材料上的效率。将结果与使用具有相似比表面积/载体材料的多孔二氧化硅进行的相应实验进行比较。此外,还评估了改性氧化锆和二氧化硅生物催化剂的储存稳定性。这些结果表明,使用这种新型氧化锆载体材料可以实现特定的固定化酶生物催化剂,其表现出与更传统的二氧化硅基材料不同的特性。此外,与其他研究人员先前描述的在类似缓冲条件下固定在有机载体材料上的相同酶的数据相比,酶-氧化锆生物催化剂的结果也显示出不同的特征。相对于目前使用的其他替代载体材料,还描述了基于氧化锆的固定化酶生物催化剂在密度和化学稳定性方面的优势。