process for the production of a reactive matrix from a coenzyme that is associated with a wasserunloeslichen traegermaterial chemically

摘要

1412647 Reactive matrix; separation of enzymes NATIONAL RESEARCH DEVELOPMENT CORP 15 Oct 1973 [17 Oct 1972] 47822/72 Headings C3H and C3U Process for the production of a reactive matrix comprising a co-enzyme chemically attached to a water-insoluble support material which comprises reacting a thio derivative of the co-enzyme, preferably in which an amino group on the co-enzyme has been replaced by a thio group with:- (a) a bifunctional organic compound having a nucleophilic group so as to couple the bifunctional organic compound to the co-enzyme and then coupling the resultant co-enzyme reaction product to a water-insoluble support material through the unreacted functional group of the bifunctional organic compound or (b) a water-insoluble support material having a plurality of pendant nucleophilic groups so as to couple the co-enzyme to the support material. The co-enzyme, which is preferably naturally occurring may be nicotinamide adenine dinucleotide (NAD), NAD- phosphate (NADP) or their reduced forms nicotinamide mononucleotide (NMN), adenosine diphosphate ribose (ADP-ribose), adenosine triphosphate (ATP), adenosine diphosphate (ADP) an adenosine monophosphate (AMP), pyridoxamine phosphate, a pterin or any other nucleoside phosphate. The co-enzyme may, alternatively, be a fragment of a naturally occurring co-enzyme having co-enzyme activity, e.g. a nucleoside containing a pusine or pyrimide ring or a phosphate containing organic molecule. The thio derivative may be a thiol, thioether, thioester, disulphide, sulphonyl halide, sulphonic acid, sulphonic ester, or alkyl sulphonyl or sulphone derivative. The bifunctional compound preferably comprises a nitrogen containing nucleophilic group, e.g. an amino group and contains a further amino, hydroxyl or ester group, for example it may be an aliphatic or aromatic diamine having 2-12 carbon atoms. The insoluble polymer support material may be a hydrophilic material such as cellulose, dextran or an agarose. Enzyme mixtures may be separated into their constituent enzymes by contacting the mixture with the insolubilized co-enzyme wherein the co-enzyme is a co-enzyme for the enzymes in the mixture and eluting the enzymes sequentially from the support.