School of Food Science, Northeast Agricultural University, Harbin 150030, China.
School of Food Science, Northeast Agricultural University, Harbin 150030, China.
Int J Biol Macromol. 2022 Sep 1;216:498-509. doi: 10.1016/j.ijbiomac.2022.06.203. Epub 2022 Jul 4.
In this study, magnetic nanoparticles (MNPs) were combined with graphene oxide (GO) to effectively improve the immobilization effect of lipase, and magnetic graphene oxide (MGON) was modified to immobilize Candida rugosa lipase (CRL) with functionalized MGON as a carrier. The optimal magnetic carrier was screened. Characterization and analysis revealed that the specific surface area of MGON was 263.75 m·g. The magnetic saturation intensities of the carrier modified with γ-ureapropyltrimethoxysilane (TMSPU), γ-aminopropyltriethoxysilane (APTES), and γ-mercaptopropyltriethoxysilane (MPTS) were 26.67, 35.1, and 43.1 emu/g, respectively. The enzyme carrying amounts of the three immobilized enzymes were 402.6, 323.4, and 288.8 mg/g, respectively. The number of binding sites of the modified carrier of TMSPU was more than that of the two other modified carriers. The optimum temperature of magnetic immobilized enzyme was 50 °C, its optimum pH was 7.0, and it had good thermal stability. The immobilized enzyme modified with TMSPU retainsed 80.28 % of the relative enzyme activity after eight times of reusability. These results demonstrated that TMSPU-modified MGON showed potential for applications involving enzyme immobilization.
在本研究中,磁性纳米粒子(MNPs)与氧化石墨烯(GO)结合,有效地提高了脂肪酶的固定化效果,并对磁性氧化石墨烯(MGON)进行了修饰,以固定化假丝酵母脂肪酶(CRL),功能化的 MGON 作为载体。筛选出最佳的磁性载体。通过对其进行表征和分析,发现 MGON 的比表面积为 263.75 m·g。经γ-脲丙基三甲氧基硅烷(TMSPU)、γ-氨丙基三乙氧基硅烷(APTES)和γ-巯丙基三乙氧基硅烷(MPTS)修饰的载体的磁饱和强度分别为 26.67、35.1 和 43.1 emu/g。三种固定化酶的酶载量分别为 402.6、323.4 和 288.8 mg/g。TMSPU 修饰载体的结合位点数多于另外两种修饰载体。经 TMSPU 修饰的磁性固定化酶的最适温度为 50°C,最适 pH 值为 7.0,具有良好的热稳定性。经 TMSPU 修饰的固定化酶在重复使用 8 次后,相对酶活保留了 80.28%。这些结果表明,TMSPU 修饰的 MGON 在酶固定化方面具有应用潜力。