Department of Medical Biochemistry, Faculty of Health Sciences with the Division of Nursing and Midwifery, Medical University of Lodz, Mazowiecka 6/8, 92-215, Lodz, Poland.
Department of Materials Technology and Chemistry, Faculty of Chemistry, University of Lodz, Pomorska 163, 90-236, Lodz, Poland.
Appl Biochem Biotechnol. 2018 Jul;185(3):717-735. doi: 10.1007/s12010-017-2682-2. Epub 2018 Jan 3.
Human catalase cDNA was cloned into a pEX-C-His vector. Purified recombinant catalase was immobilized on nanoparticles. Gold and silver nanoparticles were synthesized in a variety of sizes by chemical reduction; no agglomerates or aggregates were observed in any of the colloids during dynamic light scattering or scanning transmission electron microscopy analysis. After immobilization on gold nanoparticles, recombinant catalase activity was found to be lower than that of the same amount of enzyme in aqueous solution. However, after 10 days of storage at room temperature, the activity of catalase immobilized on gold nanoparticles (AuNPs) of 13 and 20 nm and coverage of 133% was 68 and 83% greater than catalase in aqueous solution, respectively. During 10 days of experiment, percentage activity of catalase immobilized on those gold nanoparticles was higher in comparison to CAT in aqueous solution. Catalase immobilized on silver nanoparticles did not lose activity as significantly as catalase immobilized on AuNPs. Those results confirm the ability to produce recombinant human enzymes in a bacterial expression system and its potential use while immobilized on silver or gold nanoparticles.
人源过氧化氢酶 cDNA 被克隆到 pEX-C-His 载体中。纯化的重组过氧化氢酶固定在纳米颗粒上。通过化学还原法合成了各种尺寸的金和银纳米颗粒；在动态光散射或扫描透射电子显微镜分析中，没有观察到任何胶体中的团聚体或聚集体。固定在金纳米颗粒上后，发现重组过氧化氢酶的活性低于相同量的水溶液中的酶。然而，在室温下储存 10 天后，固定在 13 和 20nm 金纳米颗粒（AuNPs）上的过氧化氢酶的活性和覆盖率分别为 133%的活性比水溶液中的过氧化氢酶分别高 68%和 83%。在 10 天的实验过程中，与水溶液中的 CAT 相比，固定在这些金纳米颗粒上的过氧化氢酶的百分比活性更高。固定在银纳米颗粒上的过氧化氢酶的活性损失并不像固定在 AuNPs 上的过氧化氢酶那样显著。这些结果证实了在细菌表达系统中生产重组人酶的能力及其在固定在银或金纳米颗粒上时的潜在用途。
