Substrate- and product-affinity resins for adenosine deaminase obtained by immobilisation of adenosine and inosine via 2',3'-cyclic acetal derivatives.
Immobilised inosine (6a) and adenosine (6c) and their 5'-phosphates have been synthesized. Reaction of the nucleosides with ethyl levulinate, followed by saponification or phosphorylation and then saponification, gave the 2',3'-O-[1-(2-carboxyethyl)ethylidene] derivatives 3 and 4 and the corresponding 5'-phosphates 2b and 2d. 6-Aminohexylagarose (5) was severally coupled to 2b, 2d, 3, and 4 through the carboxyl groups to give the polymers 6a-d. Adenosine deaminase converts 3 into 4, and 6c into 6a. The polymers can be used as affinity resins for adenosine deaminase, which is bound more strongly to 6c than to 6a. The operational capacity of 6a for adenosine deaminase is constant at 15--25 degrees, but decreases by approximately 16% from 25 degrees to 35 degrees. The resin 6a has been used to separate adenosine deaminase from mixtures containing other enzymes, for example, guanase or alcohol dehydrogenase.
D000241:Adenosine / Q000031:analogs & derivatives; D000243:Adenosine Deaminase / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D007288:Inosine / Q000031:analogs & derivatives; D009700:Nucleoside Deaminases / Q000302:isolation & purification*; D012116:Resins, Plant
D012116:Resins, Plant; D007288:Inosine; D009700:Nucleoside Deaminases; D000243:Adenosine Deaminase; D000241:Adenosine