Abstract
Indoxyl glykosider viste seg å være verdifulle og allsidige verktøy for overvåking glykosidase aktiviteter. Indoxyls blir frigitt ved enzymatisk hydrolyse, og blir hurtig oksydert, for eksempel ved atmosfærisk oksygen, i Indigo typen fargestoffer. Denne reaksjonen muliggjør rask og enkel screening in vivo uten isolering eller rensing av enzymer, samt hurtigprøver på agarplater eller i løsning (for eksempel blå-hvit screening, mikro-brønner) og blir brukt i biokjemi, histokjemi, bakteriologi og molekylær biologi. Dessverre er syntesen av slike substrater viste seg å være vanskelig, på grunn av forskjellige bireaksjoner og den lave reaktiviteten av indoksyl hydroksylfunksjon. Spesielt for glukose typen strukturer lave utbytter ble observert. Vår ny tilnærming syssels indoxylic ester som viktige mellomprodukter. Indoxylic estere med varierte erstatningsmønstre var forberedt på skalerbare trasé. Faseoverførings glycosylations med disse acceptors og peracetylatedglycosylhalogenider kan utføres under vanlige betingelser i høye utbytter. Esterspalting og påfølgende mild sølv mediert glykosylering gir de peracetylated Indoxyl glykosider i høye utbytter. Endelig avbeskyttelse utføres ifølge Zemplén.
Materials
| Name | Company | Catalog Number | Comments |
| Acetic anhydride | Grüssing | 10298 | Corrosive, flammable |
| Acetonitrile | Sigma-Aldrich | 608-001-00-3 | Harmful, flammable |
| Allyl alcohol | Aldrich | 453021 | Harmful, dangerous for the environment |
| Amberlite IR-120 H+ | Fluka | 06428 | Irritant |
| Bromoacetic acid | Merck | 802260 | Corrosive, toxic, dangerous for the environment |
| 4-Bromo-3-chloro-2-methylaniline | ABCR | AB 171687 | Irritant |
| Dichloromethane | ACROS | 326850010 | Harmful |
| Diethyl ether | Grüssing | 10274 | Harmful, extremly flammable |
| Dimethylformamide | ACROS | 348430010 | Harmful, flammable |
| Dimethylsulfoxide | Sigma-Aldrich | 41648 | |
| Ethyl acetate | Sigma-Aldrich | 607-022-00-5 | Irritant, flammable |
| Ethylenediaminetetraacetic acid | AppliChem | A1103.0500 | Irritant |
| β1,3-Galactosidase, recombinant, E. coli | Calbiochem | 345795 | |
| Hydrochloric acid | VWR | 20252.290 | Corrosive |
| Magnesium sulfate hydrate | Merck | 105885 | |
| Methanol | ACROS | 326950010 | Toxic, flammable |
| Morpholine | Janssen Chimica | 15.868.57 | Corrosive, flammable |
| Peroleum ether | Azelis | 111053 | Flammable, irritant, dangerous for the environment |
| Potassium carbonate | Grüssing | 12005 | Corrosive |
| Potassium permanganate | Grüssing | 12056 | Harmful, oxidising |
| Potassium tert-butoxide | Merck | 804918 | Corrosive, flammable |
| Pyridine | Sigma-Aldrich | 613-002-00-7 | Harmful, flammable |
| Silver acetate | Fluka | 85140 | Irritant, dangerous for the environment |
| Sodium bicarbonate | Grüssing | 12144 | Corrosive |
| Sodium hydride | Merck | 814552 | Corrosive, flammable |
| Sodium hydroxide | Riedel-de Häen | S181200 | Corrosive |
| Sodium methanolate | Merck | 806538 | Corrosive, flammable |
| Sodium sulfate | Grüssing | 12175 | |
| Tetrabutylammonium hydrogensulfate | Lancaster | 5438 | Harmful |
| Tetrahydrofurane | Sigma-Aldrich | 87371 | Harmful, flammable |
| Tetrakis (triphenylphosphine)palladium(0) | Sigma-Aldrich | 216666 | |
| Triphosgene | Fluka | 15217 | Toxic |
| Tris (hydroxymethyl)aminomethane hydrochloride | Sigma | T-3253 | Irritant |
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