CAS Key Laboratory of Chemistry of Northwestern Plant Resources, Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Lanzhou, 730000, PR China; University of Chinese Academy of Sciences, Beijing, 100049, PR China.
CAS Key Laboratory of Chemistry of Northwestern Plant Resources, Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Lanzhou, 730000, PR China.
Anal Chim Acta. 2023 Nov 1;1280:341880. doi: 10.1016/j.aca.2023.341880. Epub 2023 Oct 5.
β-Glucosidase (β-Gluco) is an enzyme that is crucial to numerous diseases, including cancer, and in sector of industries, it is used in the manufacturing of food. Measuring its enzymatic activity is critical for biomedical studies and other activities. Herein, we have developed a novel and precise fluorescent sensing method for measuring β-Gluco activity based on the production of yellow-green fluorescent quercetin-silicon nanoparticles (Q-SiNPs) produced from quercetin (QN) as a reducing agent and 3-[2-(2-aminoethyl amino) ethylamino] propyl-trimethoxy silane (AEEA) as a silane molecule. β-Gluco hydrolyzed quercetin-3-O-β-d-glucopyranoside (QO-β-DG) to produce QN, which was then used to produce Q-SiNPs. Reaction parameters, including temperature, time, buffer, pH, and probe concentration, were carefully tuned in this study. Subsequently, the fluorescence intensity was performed, showing good linearity (R = 0.989), a broad linear dynamic range between 0.5 and 12 U L, and a limit of detection (LOD) as low as 0.428 U L, which was proven by fluorescence measurements. Most importantly, various parameters were detected and characterized with or without β-Gluco. The designed probe was successively used to assess β-Gluco activity in human serum and moldy bread. However, the mathematical findings revealed recoveries for human serum ranging from 99.3 to 101.66% and for moldy bread from 100.11 to 102.5%. Additionally, Q-SiNPs were well suited to being incubated in vitro with L929 and SiHa living cells, and after using an Olympus microscope, imaging showed good fluorescence cell images, and their viability evinced minimal cytotoxicity of 77% for L929 and 88% for SiHa. The developed fluorescence biosensor showed promise for general use in diagnostic tests. Therefore, due to this outstanding sensing modality, we anticipate that this research can provide a novel schematic project for creating simple nanostructures with a suitable plan and a green synthetic option for enzyme activity and cell imaging.
β-葡萄糖苷酶（β-Gluco）是一种对包括癌症在内的许多疾病至关重要的酶，在工业领域，它被用于食品制造。测量其酶活性对于生物医学研究和其他活动至关重要。在此，我们开发了一种新颖而精确的荧光传感方法，用于测量β-葡萄糖苷酶的活性，该方法基于从槲皮素（QN）作为还原剂和 3-[2-(2-氨乙基氨基)乙基氨基]丙基三甲氧基硅烷（AEEA）作为硅烷分子生产的黄绿光致荧光槲皮素-硅纳米粒子（Q-SiNPs）的产生。β-葡萄糖苷酶将槲皮素-3-O-β-d-吡喃葡萄糖苷（QO-β-DG）水解产生 QN，然后用于生产 Q-SiNPs。在这项研究中，仔细调整了反应参数，包括温度、时间、缓冲液、pH 值和探针浓度。随后进行了荧光强度的测定，结果表明，荧光强度具有良好的线性（R=0.989），在 0.5 至 12 U L 之间具有较宽的线性动态范围，检测限（LOD）低至 0.428 U L，这通过荧光测量得到了证明。最重要的是，在有或没有β-葡萄糖苷酶的情况下检测和表征了各种参数。该设计的探针成功地用于评估人血清和霉变面包中的β-葡萄糖苷酶活性。然而，数学发现表明，人血清的回收率范围为 99.3%至 101.66%，霉变面包的回收率范围为 100.11%至 102.5%。此外，Q-SiNPs 非常适合与 L929 和 SiHa 活细胞在体外共孵育，使用奥林巴斯显微镜进行成像后，显示出良好的荧光细胞图像，其细胞活力显示出对 L929 的最小细胞毒性为 77%，对 SiHa 的最小细胞毒性为 88%。开发的荧光生物传感器有望在诊断测试中得到广泛应用。因此，由于这种出色的传感模式，我们预计这项研究可以为创建具有合适方案的简单纳米结构和酶活性和细胞成像的绿色合成选择提供一个新的方案。
