National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China.
Bioresour Technol. 2012 Dec;126:117-22. doi: 10.1016/j.biortech.2012.09.005. Epub 2012 Sep 10.
A simple and highly efficient protocol using magnetic mesoporous silica nanoparticles (MMSNPs) with metal affinity ligands was developed to directly capture laccase from Trametes versicolor fermentation broth. The Cu(2+)-chelated magnetic mesoporous silica nanoparticles (MMSNPs-Cu(2+)) with pore sizes ranging from 3.6 to 27.1 nm exhibited size selectivity on laccase capture from the fermentation broth, and the MMSNPs-Cu(2+) with an average pore size of 14.5 nm provided 60.6-fold purification of laccase and 114.6% recovery yield of enzyme activity. Both size selectivity of the MMSNPs and affinity of the chelated metal ion resulted in high laccase capture efficiency from the fermentation broth. The most efficient MMSNPs-Cu(2+) demonstrated no significant loss in laccase capture effectiveness following 10 reuse cycles. This simple and efficient strategy has the potential to be used for the robust and inexpensive preparation of purified laccase at the industrial scale.
开发了一种使用带有金属亲和配体的磁性介孔硅纳米粒子（MMSNPs）的简单高效的方案，以直接从变色栓菌发酵液中捕获漆酶。Cu(2+)-螯合的介孔磁性硅纳米粒子（MMSNPs-Cu(2+)）的孔径范围为 3.6 至 27.1nm，对发酵液中的漆酶具有尺寸选择性，平均孔径为 14.5nm 的 MMSNPs-Cu(2+)提供了 60.6 倍的漆酶纯化倍数和 114.6%的酶活回收率。MMSNPs 的尺寸选择性和螯合金属离子的亲和性都导致了发酵液中漆酶的高效捕获。最有效的 MMSNPs-Cu(2+)在经过 10 次重复使用循环后，在漆酶捕获效果上没有明显的损失。这种简单高效的策略有望用于在工业规模上制备稳健且廉价的纯化漆酶。
