Immobilized capillary adenosine deaminase microreactor for inhibitor screening in natural extracts by capillary electrophoresis.
A novel strategy for the preparation of in-column adenosine deaminase (ADA) microreactor and rapid screening of enzyme inhibitors in natural extracts was demonstrated. In this approach, ADA was encapsulated in anionic polyelectrolyte alginate that was immobilized on the surface of fused-silica capillary via ionic binding technique with cationic polyelectrolyte polyethylenimine (PEI). On-line enzyme inhibition study was performed by capillary electrophoresis (CE). The substrate and product were baselined separated within 75s. The enzyme activity was determined by the quantification of peak area of the product. Enzyme inhibition can be read out directly from the reduced peak area of the product in comparison with a reference electropherogram obtained in the absence of any inhibitor. The inhibition percentage was used to evaluate relative activity of ADA microreactor. A known ADA inhibitor, erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) was employed as a model compound for the validation of the inhibitor screening method, and the screening of ADA inhibitor in 19 traditional Chinese herbal medicines was performed.
Ji
Xiaowen
X
The Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources Ministry of Education, College of Chemistry and Chemical Engineering, Guangxi Normal University, Guilin 541004, China.
Ye
Fanggui
F
Lin
Pingtan
P
Zhao
Shulin
S
D000243:Adenosine Deaminase / Q000378:metabolism*; D058892:Adenosine Deaminase Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D019149:Bioreactors; D019075:Electrophoresis, Capillary / Q000379:methods*; D013379:Substrate Specificity
D058892:Adenosine Deaminase Inhibitors; D000243:Adenosine Deaminase