Biomolecular Interaction Centre and School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, New Zealand.
Biotechnol Prog. 2011 Mar-Apr;27(2):360-7. doi: 10.1002/btpr.518. Epub 2010 Nov 23.
Organophosphate hydrolase has potential as a bioremediation and chemical detoxification enzyme, but the problems of reusability and stability need to be addressed to use this enzyme on an industrial scale. Immobilizing the enzyme to a nanoscaffold may help to solve these problems. Amyloid fibrils generated from insulin and crystallin provided a novel nanoscaffold for the immobilization of organophosphate hydrolase, using glutaraldehyde as the crosslinking reagent. Electrophoretic, centrifugation, and temperature stability experiments, together with transmission electron microscopy were undertaken to verify that crosslinking had successfully occurred. The resulting fibrils remained active towards the substrate paraoxon and when immobilized to the insulin amyloid fibrils, the enzyme exhibited a significant (∼ 300%) increase in the relative temperature stability at 40, 45, and 50°C (as measured by comparing the initial enzyme activity to the activity remaining after heating), compared to free enzyme. This confirms that amyloid fibrils could provide a new type of nanoscaffold for enzyme immobilization.
有机磷水解酶具有生物修复和化学解毒酶的潜力，但需要解决可重复使用性和稳定性的问题，才能在工业规模上使用这种酶。将酶固定在纳米支架上可能有助于解决这些问题。来自胰岛素和晶体蛋白的淀粉样纤维为有机磷水解酶的固定提供了一种新颖的纳米支架，使用戊二醛作为交联试剂。进行电泳、离心和温度稳定性实验，以及透射电子显微镜观察，以验证交联是否成功发生。结果表明，这些纤维仍然对底物对氧磷具有活性，并且当固定在胰岛素淀粉样纤维上时，与游离酶相比，酶在 40、45 和 50°C 时的相对温度稳定性显著提高（∼300%）（通过比较初始酶活性与加热后剩余的活性来衡量）。这证实了淀粉样纤维可以为酶固定提供一种新型的纳米支架。
