Araújo Rita, Silva Carla, Machado Raul, Casal Margarida, Cunha António M, Rodriguez-Cabello José Carlos, Cavaco-Paulo Artur
Department of Biology, Centre of Molecular and Environmental Biology, University of Minho, Braga, Portugal.
Biomacromolecules. 2009 Jun 8;10(6):1655-61. doi: 10.1021/bm9002943.
One of the goals of protein engineering is to tailor the structure of enzymes to optimize industrial bioprocesses. In the present work, we present the construction of a novel high molecular weight subtilisin, based on the fusion of the DNA sequences coding for Bacillus subtilis prosubtilisin E and for an elastin-like polymer (ELP). The resulting fusion protein was biologically produced in Escherichia coli , purified and used for wool finishing assays. When compared to the commercial protease Esperase, the recombinant subtilisinE-VPAVG(220) activity was restricted to the cuticle of wool, allowing a significant reduction of pilling, weight loss and tensile strength loss of wool fibers. Here we report, for the first time, the microbial production of a functionalized high molecular weight protease for controlled enzymatic hydrolysis of wool surface. This original process overcomes the unrestrained diffusion and extended fiber damage which are the major obstacles for the use of proteases for wool finishing applications.
蛋白质工程的目标之一是调整酶的结构以优化工业生物过程。在本研究中,我们基于编码枯草芽孢杆菌枯草杆菌蛋白酶原E和弹性蛋白样聚合物(ELP)的DNA序列的融合,构建了一种新型的高分子量枯草杆菌蛋白酶。所得融合蛋白在大肠杆菌中进行生物生产、纯化,并用于羊毛整理试验。与市售蛋白酶Esperase相比,重组枯草杆菌蛋白酶E-VPAVG(220)的活性仅限于羊毛的表皮,可显著减少羊毛纤维的起球、失重和拉伸强度损失。在此,我们首次报道了用于羊毛表面可控酶促水解的功能化高分子量蛋白酶的微生物生产。这一原创工艺克服了无限制扩散和纤维过度损伤的问题,而这些问题是蛋白酶用于羊毛整理应用的主要障碍。