Department of Biocatalysis, Institute of Catalysis, CSIC, Cantoblanco, 28049 Madrid, Spain.
Applied Biotechnology Group, Faculty of Biomedical and Health Sciences, Universidad Europea de Madrid, Urbanización El Bosque, Villaviciosa de Odón, 28670 Madrid, Spain.
Int J Mol Sci. 2019 Apr 2;20(7):1627. doi: 10.3390/ijms20071627.
Unspecific peroxygenases (UPOs) are highly promiscuous biocatalyst with self-sufficient mono(per)oxygenase activity. A laboratory-evolved UPO secreted by yeast was covalently immobilized in activated carriers through one-point attachment. In order to maintain the desired orientation without compromising the enzyme's activity, the S221C mutation was introduced at the surface of the enzyme, enabling a single disulfide bridge to be established between the support and the protein. Fluorescence confocal microscopy demonstrated the homogeneous distribution of the enzyme, regardless of the chemical nature of the carrier. This immobilized biocatalyst was characterized biochemically opening an exciting avenue for research into applied synthetic chemistry.
非特异性过氧化物酶（UPO）是一种高度混杂的生物催化剂，具有自给自足的单（过）氧合酶活性。通过单点附着，酵母分泌的实验室进化的 UPO 被共价固定在活化载体中。为了在不损害酶活性的情况下保持所需的取向，在酶的表面引入了 S221C 突变，从而在载体和蛋白质之间建立了单个二硫键。荧光共焦显微镜显示了酶的均匀分布，而与载体的化学性质无关。这种固定化生物催化剂具有生物化学特性，为应用合成化学的研究开辟了令人兴奋的途径。
