Instrumental Analysis Center, Dalian Polytechnic University, Dalian 116034, China; School of Biological Engineering, Dalian Polytechnic University, Dalian 116034, China.
School of Biological Engineering, Dalian Polytechnic University, Dalian 116034, China.
Enzyme Microb Technol. 2021 Dec;152:109935. doi: 10.1016/j.enzmictec.2021.109935. Epub 2021 Oct 22.
Enzymes are particularly attractive as biocatalysts for the green synthesis of chemicals and pharmaceuticals. However, the traditional enzyme purification and separation process is complex and inefficient, which limits the wide application of enzyme catalysis. In this paper, an efficient strategy for enzyme purification and immobilization in one step is proposed. A novel poly (ionic liquid)-styrene microsphere is prepared by molecular design and synthesis for adsorbing and purifying high activity lipase from fermentation broth directly. By optimizing the surface morphologies and charge of the microspheres, the enzyme loading is significantly improved. In order to further stabilize the catalytic environment of lipase, the resulting lipase/poly (ionic liquid)-styrene microspheres are immobilized in physical crosslinking hydrogel to obtain a complex lipase catalytic system, which can be prepared into various shapes according to the requirements of catalytic environment. In the actual catalytic reaction process, this complex lipase catalytic system exhibits excellent catalytic activity (6314.69 ± 21.27 U mg) and good harsh environment tolerance compared with the lipase fermentation broth (1672.87 ± 36.68 U mg). Under the condition of cyclic catalysis, the complex lipase catalytic system shows the outstanding reusability (After 8 cycles the enzymatic activity is still higher than that of the lipase fermentation broth) and is easily separated from the products.
酶作为化学和药物绿色合成的生物催化剂具有独特的吸引力。然而，传统的酶纯化和分离过程复杂且效率低下，限制了酶催化的广泛应用。本文提出了一种在一步中进行酶纯化和固定化的有效策略。通过分子设计和合成，制备了一种新型的聚离子液体-苯乙烯微球，可直接从发酵液中吸附和纯化高活性脂肪酶。通过优化微球的表面形态和电荷，显著提高了酶的负载量。为了进一步稳定脂肪酶的催化环境，将得到的脂肪酶/聚离子液体-苯乙烯微球固定在物理交联水凝胶中，得到一种复杂的脂肪酶催化体系，可根据催化环境的要求制备成各种形状。在实际催化反应过程中，与脂肪酶发酵液（1672.87 ± 36.68 U mg）相比，该复合脂肪酶催化体系表现出优异的催化活性（6314.69 ± 21.27 U mg）和良好的恶劣环境耐受性。在循环催化条件下，该复合脂肪酶催化体系表现出优异的可重复使用性（经过 8 次循环后，酶活性仍高于脂肪酶发酵液），并且易于与产物分离。
