Department of Biology, Faculty of Science, Nong Lam University, Ho Chi Minh City, 71300, Vietnam.
Graduate School of Biotechnology, College of Life Science, Kyung Hee University, Yongin-si, Gyeonggi-do, 17104, Republic of Korea.
Arch Microbiol. 2024 Oct 3;206(11):423. doi: 10.1007/s00203-024-04148-4.
Minor ginsenosides produced by β-glucosidase are interesting biologically and pharmacologically. In this study, new ginsenoside-hydrolyzing glycosidase from Furfurilactobacillus rossiae DCYL3 was cloned and expressed in Escherichia coli strain BL21. The enzyme converted Rb1 and Gyp XVII into Rd and compound K following the pathways: Rb1→Rd and Gyp XVII→F2→CK, respectively at optimal condition: 40 °C, 15 min, and pH 6.0. Furthermore, we examined the cytotoxicity, NO production, ROS generation, and gene expression of Gynostemma extract (GE) and bioconverted Gynostemma extract (BGE) in vitro against A549 cell lines for human lung cancer and macrophage RAW 264.7 cells for antiinflammation, respectively. As a result, BGE demonstrated significantly greater toxicity than GE against lung cancer at a dose of 500 µg/mL but in normal cells showed lower toxicity. Then, we indicated an enhanced generation of ROS, which may be boosting cancer cell toxicity. By blocking the intrinsic way, BGE increased p53, Bax, Caspase 3, 9, and while Bcl2 is decreased. At 500 µg/mL, the BGE sample was less toxic in normal cells and decreased the LPS-treated NO and ROS level to reduce inflammation. In addition, BGE inhibited the expression of pro-inflammatory genes COX-2, iNOS, IL-6, and IL-8 in RAW 264.7 cells than the sample of GE. In conclusion, FrBGL3 has considerable downstream applications for high-yield, low-cost, effective manufacture of minor ginsenosides. Moreover, the study's findings imply that BGE would be potential materials for anti-cancer and anti-inflammatory agent after consideration of future studies.
β-葡萄糖苷酶产生的低聚糖具有有趣的生物学和药理学特性。本研究从罗斯氏乳杆菌（Furfurilactobacillus rossiae）DCYL3 中克隆并在大肠杆菌 BL21 菌株中表达了新的人参皂苷水解糖苷酶。该酶在最佳条件下（40°C、15 分钟、pH6.0）分别通过以下途径将 Rb1 和 Gyp XVII 转化为 Rd 和化合物 K：Rb1→Rd 和 Gyp XVII→F2→CK。此外，我们分别在体外研究了人参提取物（GE）和生物转化的人参提取物（BGE）对人肺癌 A549 细胞系的细胞毒性、NO 产生、ROS 生成和基因表达，以及对巨噬细胞 RAW 264.7 细胞的抗炎作用。结果表明，BGE 在 500µg/mL 剂量下对肺癌的毒性明显大于 GE，但在正常细胞中毒性较低。然后，我们表明 ROS 的生成增强，这可能会增强癌细胞的毒性。通过阻断内在途径，BGE 增加了 p53、Bax、Caspase 3、9，而 Bcl2 减少。在 500µg/mL 时，BGE 样品在正常细胞中的毒性较低，并降低 LPS 处理的 NO 和 ROS 水平以减轻炎症。此外，BGE 抑制了 RAW 264.7 细胞中促炎基因 COX-2、iNOS、IL-6 和 IL-8 的表达，其抑制作用强于 GE 样品。总之，FrBGL3 具有相当大的下游应用潜力，可用于高效、低成本、有效制造低聚糖。此外，考虑到未来的研究，研究结果表明，BGE 可能是抗癌和抗炎药物的潜在材料。
