Highly Efficient Preparation of Resorufin-beta-D-glucuronide
文献类型：期刊论文
| 作者 | Wang Xinxin1; Lu Xia3; Ge Guangbo3; Feng Lei3; Xin Hong3; Li Yaoguang3; Cao Yunfeng3; Han Guanying2; Guo Bin2 |
| 刊名 | chemical journal of chinese universities-chinese
 |
| 出版日期 | 2015-07-10 |
| 卷号 | 36期号:7页码:1321-1327 |
| 关键词 | Resorufin Resorufin-beta-O-glucuronide Solid-phase extraction beta-Glucuronidase Inhibitor screening |
| 英文摘要 | resorufin-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标题词 | science & technology ; physical sciences |
| 类目[WOS] | chemistry, multidisciplinary |
| 研究领域[WOS] | chemistry |
| 关键词[WOS] | inhibition ; expression ; substrate |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000358472100013 |
| 公开日期 | 2016-05-09 |
| 源URL | [http://cas-ir.dicp.ac.cn/handle/321008/146416]  |
| 专题 | 大连化学物理研究所_中国科学院大连化学物理研究所 |
| 作者单位 | 1.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 |
| 推荐引用方式 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. |
入库方式： OAI收割
来源：大连化学物理研究所
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