Institute of Agro-food Technology, Jilin Academy of Agricultural Sciences (Northeast Agricultural Research Center of China), Changchun 130033, P. R. China.
School of Pharmaceutical Sciences, Changchun University of Chinese Medicine, Changchun 130117, P.R. China.
J Appl Microbiol. 2024 Jul 2;135(7). doi: 10.1093/jambio/lxae180.
Microbial transformation to modify saponins and enhance their biological activities has received increasing attention in recent years. This study aimed to screen the strain that can biotransform notoginsenoside R1, identify the product and study its biological activity.
A lactic acid bacteria strain S165 with glycosidase-producing activity was isolated from traditional Chinese fermented foods, which was identified and grouped according to API 50 CHL kit and 16S rDNA sequence analysis. Subsequently, notoginsenoside R1 underwent a 30-day fermentation period by the strain S165, and the resulting products were analyzed using High-performance liquid chromatography (HPLC), Ultra-performance liquid chromatography (UPLC)-mass spectrometry (MS)/MS, and 13C-Nuclear magnetic resonance (NMR) techniques. Employing a model of Lipopolysaccharide (LPS)-induced damage to Caco-2 cells, the damage of Caco-2 cells was detected by Hoechst 33 258 staining, and the activity of notoginsenoside R1 biotransformation product was investigated by CCK-8 and western blotting assay. The strain S165 was identified as Lactiplantibacillus plantarum and was used to biotransform notoginsenoside R1. Through a 30-day biotransformation, L. plantarum S165 predominantly converts notoginsenoside R1 into 3β,12β-dihydroxydammar-(E)-20(22),24-diene-6-O-β-D-xylopyranosyl-(1→2)-β-D-glucopyranoside, temporarily named notoginsenoside T6 (NGT6) according to HPLC, UPLC-MS/MS, and 13C-NMR analysis. Results from CCK-8 and Hoechst 33258 staining indicated that the ability notoginsenoside T6 to alleviate the intestinal injury induced by LPS in the Caco-2 cell was stronger than that of notoginsenoside R1. In addition, Western blotting result showed that notoginsenoside T6 could prevent intestinal injury by protecting tight junction proteins (Claudin-1, Occludin, and ZO-1).
Notoginsenoside R1 was biotransformed into the notoginsenoside T6 by L. plantarum S165, and the biotransformed product showed an enhanced intestinal protective effect in vitro.
近年来，微生物转化修饰皂苷并提高其生物活性的方法受到了越来越多的关注。本研究旨在筛选能够生物转化人参皂苷 R1 的菌株，鉴定产物并研究其生物学活性。
从传统的中国发酵食品中分离出一株具有糖苷酶产生活性的乳酸菌 S165，根据 API 50 CHL 试剂盒和 16S rDNA 序列分析对其进行鉴定和分组。随后，采用 S165 菌株对人参皂苷 R1 进行 30 天发酵，采用高效液相色谱（HPLC）、超高效液相色谱（UPLC）-质谱（MS）/MS 和 13C-核磁共振（NMR）技术对产物进行分析。采用脂多糖（LPS）诱导的 Caco-2 细胞损伤模型，通过 Hoechst 33258 染色检测 Caco-2 细胞的损伤，通过 CCK-8 和 Western blotting 检测人参皂苷 R1 生物转化产物的活性。S165 菌株被鉴定为植物乳杆菌，用于生物转化人参皂苷 R1。经过 30 天的生物转化，L. plantarum S165 主要将人参皂苷 R1 转化为 3β，12β-二羟基达玛-（E）-20（22），24-二烯-6-O-β-D-木吡喃糖基-（1→2）-β-D-吡喃葡萄糖苷，根据 HPLC、UPLC-MS/MS 和 13C-NMR 分析，暂时命名为人参皂苷 T6（NGT6）。CCK-8 和 Hoechst 33258 染色结果表明，人参皂苷 T6 减轻 LPS 诱导的 Caco-2 细胞肠损伤的能力强于人参皂苷 R1。此外，Western blotting 结果表明，人参皂苷 T6 可以通过保护紧密连接蛋白（Claudin-1、Occludin 和 ZO-1）来预防肠道损伤。
人参皂苷 R1 被植物乳杆菌 S165 生物转化为人参皂苷 T6，其生物转化产物在体外显示出增强的肠道保护作用。
