Proteolytic Enzyme Engineering: A Tool for Wool

被引:24
作者
Araujo, Rita [1 ,2 ]
Silva, Carla [2 ]
Machado, Raul [1 ]
Casal, Margarida [1 ]
Cunha, Antonio M. [3 ]
Carlos Rodriguez-Cabello, Jose [4 ]
Cavaco-Paulo, Artur [2 ]
机构
[1] Univ Minho, Dept Biol, CBMA Ctr Mol & Environm Biol, P-4710057 Braga, Portugal
[2] Univ Minho, Dept Text Engn, Ctr Sci & Textile Engn 2C2T, P-4800058 Guimaraes, Portugal
[3] Univ Minho, Dept Polymer Engn, P-4800058 Guimaraes, Portugal
[4] Univ Valladolid, ETSII, BIOFORGE Res Grp, Dpto Fis Mat Condensada, E-47011 Valladolid, Spain
关键词
RECOMBINANT PROTEINS; ESCHERICHIA-COLI; MOLECULAR-BASIS; POLYMER; PURIFICATION; SUBTILISIN; PROTEASES; FUSION;
D O I
10.1021/bm9002943
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:1655 / 1661
页数:7
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