In the present study, a facile functionalization of magnetic nanoparticles has been described for the immobilization of enzyme that offers many advantages for reuse and excellent efficiencies. The magnetic gold nanocomposites have been fabricated for the successful immobilization of an industrially important enzyme "papain". For immobilization of papain on magnetic gold nanocomposites, magnetic nanoparticles were modified with 3-(mercaptopropyl) trimethoxy silane (MPTS). Further, the citrate stabilized gold nanoparticles were chemisorbed on these thiol coated magnetic nanoparticles to fabricate the desired magnetic gold nanocomposites. Papain containing net positive charge (isoelectric point of papain=8.75) in PBS buffer (pH 7.4) has immobilized on the surface of the negatively charged magnetic gold nanocomposites through the ionic or electrostatic interaction. The Michaelis-Menten kinetic constant (K(m)) and maximum reaction velocity (V(max)) for free papain were 0.236×10(5) g ml(-1) and 4.08 g ml(-1)/s respectively whereas for immobilized papain, K(m) and V(max) values were 0.308×10(5) g ml(-1) and 5.4 g ml(-1)/s respectively. The loading amount of papain on magnetic gold nanocomposites was 54 mg/g support and the activity recovery of the immobilized papain reached to 47 (±5)% compared to native papain. The main advantage of this papain nanobiocatalyst is the easy isolation of enzyme from the reaction medium.

译文

:在本研究中,已描述了磁性纳米粒子的简便功能化,用于固定化酶,该酶具有许多可重复使用的优点和出色的效率。已经制造出磁性金纳米复合材料,以成功固定工业上重要的酶“木瓜蛋白酶”。为了将木瓜蛋白酶固定在磁性金纳米复合材料上,用3-(巯基丙基)三甲氧基硅烷(MPTS)修饰了磁性纳米粒子。此外,将柠檬酸盐稳定的金纳米颗粒化学吸附在这些硫醇涂覆的磁性纳米颗粒上,以制造所需的磁性金纳米复合材料。在PBS缓冲液(pH 7.4)中含有净正电荷的木瓜蛋白酶(木瓜蛋白酶的等电点= 8.75)已通过离子或静电相互作用固定在带负电荷的磁性金纳米复合材料的表面上。游离木瓜蛋白酶的Michaelis-Menten动力学常数(K(m))和最大反应速度(V(max))分别为0.236×10(5)g ml(-1)和4.08 g ml(-1)/ s固定化木瓜蛋白酶的K(m)和V(max)值分别为0.308×10(5)g ml(-1)和5.4 g ml(-1)/ s。木瓜蛋白酶在磁性金纳米复合材料上的负载量为54 mg / g载体,与天然木瓜蛋白酶相比,固定化木瓜蛋白酶的活性回收率达到47(±5)%。这种木瓜蛋白酶纳米生物催化剂的主要优点是易于从反应介质中分离出酶。

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