Bioengineering and Environmental Centre, Indian Institute of Chemical Technology, Hyderabad 500007, India.
Appl Biochem Biotechnol. 2010 Apr;160(7):1888-95. doi: 10.1007/s12010-009-8726-5. Epub 2009 Aug 16.
In the present investigation, impact of nickel-impregnated silica paramagnetic particles (NSP) as biocatalyst immobilization matrices was investigated. These nanoparticles were synthesized by sol-gel route using a nonionic surfactant block co polymer [poly (ethylene glycol)-block-poly-(propylene glycol)-block-poly (ethylene glycol)]. Diastase enzyme was immobilized on these particles (enzyme-impregnated NSP) as model enzyme and characterized using Fourier-transform infrared spectroscopy and X-ray crystallography. Analysis of enzyme-binding nature with these nanoparticles at different physiological conditions revealed that binding pattern and activity profile varied with the pH of the reaction mixture. The immobilized enzyme was further characterized for its biocatalytic activity with respect to kinetic properties such as Km and Vmax and compared with free enzyme. Paramagnetic nanoparticle-immobilized enzyme showed more affinity for substrate compared to free one. The nature of silica and nickel varied from amorphous to crystalline nature and vice versa upon immobilization of enzyme. To the best of our knowledge, this is the first report of its kind for change of nature from one form to other under normal temperatures upon diastase interaction with NSP.
在本研究中,考察了负载镍的二氧化硅超顺磁纳米颗粒(NSP)作为生物催化剂固定化基质的影响。这些纳米颗粒是通过溶胶-凝胶法使用非离子表面活性剂嵌段共聚物[聚(乙二醇)-嵌段-聚(丙二醇)-嵌段-聚(乙二醇)]合成的。将淀粉酶固定在这些颗粒上(酶负载的 NSP)作为模型酶,并使用傅里叶变换红外光谱和 X 射线晶体学进行了表征。在不同生理条件下分析这些纳米颗粒与酶的结合特性表明,结合模式和活性谱随反应混合物的 pH 值而变化。进一步对固定化酶进行了生物催化活性的特性分析,包括动力学性质如 Km 和 Vmax,并与游离酶进行了比较。与游离酶相比,固定化酶对底物表现出更高的亲和力。固定化酶后,二氧化硅和镍的性质从无定形变为结晶态,反之亦然。据我们所知,这是在与 NSP 相互作用时,在正常温度下酶从一种形式转变为另一种形式的首例报道。