Enzymatic preparation of silybin phase II metabolites: sulfation using aryl sulfotransferase from rat liver

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F13%3A10191570" target="_blank" >RIV/00216208:11310/13:10191570 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61388971:_____/13:00421796

Výsledek na webu

<a href="http://dx.doi.org/10.1007/s00253-013-4794-0" target="_blank" >http://dx.doi.org/10.1007/s00253-013-4794-0</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s00253-013-4794-0" target="_blank" >10.1007/s00253-013-4794-0</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Enzymatic preparation of silybin phase II metabolites: sulfation using aryl sulfotransferase from rat liver

Popis výsledku v původním jazyce

Aryl sulfotransferase IV (AstIV) from rat liver was overexpressed in Escherichia coli and purified to homogeneity. Using the produced mammalian liver enzyme, sulfation-the Phase II conjugation reaction-of optically pure silybin diastereoisomers (silybinA and B) was tested. As a result, silybin B was sulfated yielding 20-O-silybin B sulfate, whereas silybin A was completely resistant to the sulfation reaction. Milligram-scale sulfation of silybin B was optimized employing resting E. coli cells producingAstIV, thus avoiding the use of expensive 3'-phosphoadenosine-5'-phosphate cofactor and laborious enzyme purification. Using this approach, we were able to reach 48 % conversion of silybin B into its 20-sulfate within 24 h. The sulfated product was isolated by solid phase extraction and its structure was characterized by HRMS and NMR. Sulfation reaction of silybin appeared strictly stereoselective; only silybin B was sulfated by AstIV.

Název v anglickém jazyce

Enzymatic preparation of silybin phase II metabolites: sulfation using aryl sulfotransferase from rat liver

Popis výsledku anglicky

Aryl sulfotransferase IV (AstIV) from rat liver was overexpressed in Escherichia coli and purified to homogeneity. Using the produced mammalian liver enzyme, sulfation-the Phase II conjugation reaction-of optically pure silybin diastereoisomers (silybinA and B) was tested. As a result, silybin B was sulfated yielding 20-O-silybin B sulfate, whereas silybin A was completely resistant to the sulfation reaction. Milligram-scale sulfation of silybin B was optimized employing resting E. coli cells producingAstIV, thus avoiding the use of expensive 3'-phosphoadenosine-5'-phosphate cofactor and laborious enzyme purification. Using this approach, we were able to reach 48 % conversion of silybin B into its 20-sulfate within 24 h. The sulfated product was isolated by solid phase extraction and its structure was characterized by HRMS and NMR. Sulfation reaction of silybin appeared strictly stereoselective; only silybin B was sulfated by AstIV.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

S - Specificky vyzkum na vysokych skolach<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2013

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Applied Microbiology and Biotechnology

ISSN

0175-7598

e-ISSN

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Svazek periodika

97

Číslo periodika v rámci svazku

24

Stát vydavatele periodika

FR - Francouzská republika

Počet stran výsledku

8

Strana od-do

10391-10398

Kód UT WoS článku

000327495000014

EID výsledku v databázi Scopus

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