Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210046, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Jun 1;898:95-100. doi: 10.1016/j.jchromb.2012.04.024. Epub 2012 Apr 25.
In this paper, rutin was metabolized by human intestinal bacteria and five isolated strains including Bacillus sp. 52, Bacteroides sp. 45, 42, 22 and Veillonella sp. 32, the metabolites were identified using ultra performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS). As a result, Bacillus sp. 52 and Bacteroides sp. 45 could metabolize rutin to quercetin 3-O-glucoside and leucocyanidin. Bacteroides sp. 42 and Veillonella sp. 32 could convert rutin to leucocyanidin. Bacteroides sp. 22 could hydrolyze rutin to quercetin-3-O-glucoside. In order to further explain the metabolism pathway of rutin, the β-D-glucosidase and α-L-rhamnosidase activities of five strains were determined. Bacteroides sp. 22 could produce α-L-rhamnosidase but did not produce β-D-glucosidase or β-D-glucosidase activity was too low to be detected. The other four strains all demonstrated α-L-rhamnosidase and β-D-glucosidase activities. Furthermore, α-L-rhamnosidase and β-D-glucosidase activities of Veillonella sp. 32 and Bacteroides sp. 42 were higher than those of Bacteroides sp. 45 and Bacillus sp. 52. Based on these results, we can propose the deglycosylated rout of rutin: rutin was metabolized to be quercetin-3-O-glucoside by α-L-rhamnosidase produced from these bacteria, thereafter, quercetin-3-O-glucoside was further metabolized by β-D-glucosidase to form leucocyanidin. Because of the higher enzyme activity in Veillonella sp. 32 and Bacteroides sp. 42, quercetin-3-O-glucoside was completely metabolized to leucocyanidin by these two bacteria. Due to the lack of β-D-glucosidase activity, Bacteroides sp. 22 could not further metabolize quercetin-3-O-glucoside to leucocyanidin. This study will be helpful for understanding the deglycosylated rout of rutin and the role of different intestinal bacteria on the metabolism of natural compounds.
本文利用超高效液相色谱/四极杆飞行时间质谱联用技术（UPLC-Q-TOF/MS）对肠道细菌（包括芽孢杆菌 52 号、拟杆菌 45 号、42 号、22 号和韦荣球菌 32 号）代谢芦丁的产物进行了鉴定。结果表明，芽孢杆菌 52 号和拟杆菌 45 号可以将芦丁代谢为槲皮素 3-O-葡萄糖苷和莱菔氰定。拟杆菌 42 号和韦荣球菌 32 号可以将芦丁转化为莱菔氰定。拟杆菌 22 号可以水解芦丁生成槲皮素 3-O-葡萄糖苷。为了进一步阐明芦丁的代谢途径，测定了 5 株菌的β-D-葡萄糖苷酶和α-L-鼠李糖苷酶活性。拟杆菌 22 号可以产生α-L-鼠李糖苷酶，但不产生β-D-葡萄糖苷酶或β-D-葡萄糖苷酶活性太低而无法检测。其他 4 株菌均表现出α-L-鼠李糖苷酶和β-D-葡萄糖苷酶活性。此外，韦荣球菌 32 号和拟杆菌 42 号的α-L-鼠李糖苷酶和β-D-葡萄糖苷酶活性均高于拟杆菌 45 号和芽孢杆菌 52 号。基于这些结果，我们可以提出芦丁的去糖基化途径：芦丁首先被这些细菌产生的α-L-鼠李糖苷酶代谢生成槲皮素 3-O-葡萄糖苷，然后，槲皮素 3-O-葡萄糖苷再被β-D-葡萄糖苷酶进一步代谢生成莱菔氰定。由于韦荣球菌 32 号和拟杆菌 42 号的酶活性较高，这两种细菌可将槲皮素 3-O-葡萄糖苷完全代谢为莱菔氰定。由于缺乏β-D-葡萄糖苷酶活性，拟杆菌 22 号无法进一步将槲皮素 3-O-葡萄糖苷代谢为莱菔氰定。本研究有助于理解芦丁的去糖基化途径以及不同肠道细菌对天然化合物代谢的作用。
