Highly Efficient Preparation of Resorufin-beta-D-glucuronide
Wang Xinxin1; Lu Xia3; Ge Guangbo3; Feng Lei3; Xin Hong3; Li Yaoguang3; Cao Yunfeng3; Han Guanying2; Guo Bin2
KeywordResorufin Resorufin-beta-o-glucuronide Solid-phase Extraction Beta-glucuronidase Inhibitor Screening
Indexed BySCI
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Multidisciplinary
WOS Research AreaChemistry
AbstractResorufin-beta-O-glucuronide(KEG) is a substrate of the beta-glucuronidase enzyme which exhibited almost no fluorescence, upon addition of beta-glucuronidase a high fluorescent compound resorufin will be released. Resorufin, is a fluorescent compound possessing good optical properties. On the basis of the off-on type fluorescent reaction, REG can be served as the good fluorescent substrate of beta-glucuronidase. However, commercial available KEG is rather expensive due to the difficulty for preparation of REG and the studies on REG are very limited. This study aimed to adopt the mild biosynthesis approach to efficiently prepare REG, based on resorufin can be extensively metabolized to KEG by UDP-glucuronosyltransferases(UGTs). REG was prepared using resorufin as the starting material and liver microsomes from cynomolgus monkeys(CyLM) as the enzyme source. Resorufin(RE, 100 mu mol/L) was incubated in Tris-HCl(50 mmol/L, pH = 7.4, with 1% DMSO) with CyLM(0.5 mg/mL) at 37 degrees C for 4 h. After the optimization of the incubation conditions, the conversion of REG was more than 80%. A unique solid-phase extraction column(SPE) packed with C18WAX was used to enrich and purify the product KEG with high yield. The product was then identified by both LC-MS and NMR techniques. Finally, a high throughput screening method for beta-glucuronidase inhibitors was well established, based on the accurate kinetic parameters of REG towards beta-glucuronidase and the absorption and emission spectrum of REG.
WOS IDWOS:000358472100013
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Document Type期刊论文
Affiliation1.Liaoning Med Univ, Dept Med, Jinzhou 121000, Peoples R China
2.Liaoning Med Univ, Affiliated Hosp 1, Jinzhou 121000, Peoples R China
3.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
Recommended Citation
GB/T 7714
Wang Xinxin,Lu Xia,Ge Guangbo,et al. Highly Efficient Preparation of Resorufin-beta-D-glucuronide[J]. CHEMICAL JOURNAL OF CHINESE UNIVERSITIES-CHINESE,2015,36(7):1321-1327.
APA Wang Xinxin.,Lu Xia.,Ge Guangbo.,Feng Lei.,Xin Hong.,...&Guo Bin.(2015).Highly Efficient Preparation of Resorufin-beta-D-glucuronide.CHEMICAL JOURNAL OF CHINESE UNIVERSITIES-CHINESE,36(7),1321-1327.
MLA Wang Xinxin,et al."Highly Efficient Preparation of Resorufin-beta-D-glucuronide".CHEMICAL JOURNAL OF CHINESE UNIVERSITIES-CHINESE 36.7(2015):1321-1327.
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