- Patent JPS5631950

Document Type and Number:

Japanese Patent JPS5631950

Kind Code:

B1

Abstract:

1357861 Carrier-bound proteins; polymerizing enzyme compounds BOEHRINGER MANNHEIM GmbH 7 June 1972 [9 June 1971] 26556/72 Headings C3H and C3P Carrier-bound proteins (enzymes) are produced by reacting the protein in aqueous solution with a compound containing at least one epoxy group and at least one further functional group capable of producing a bond with a carrier substance and then reacting this further group with the carrier substance. The carrier is preferably produced within the aqueous reaction solution by the polymerization of a monomer or monomer mixture. Thus, the reaction of the protein with the epoxy compound may take place in the presence of a polymerizable monomer, whereafter polymerization is carried out with the polymerizing in of the epoxy compound-protein product. Alternatively, the polymerizable monomer may be added and polymerized after the epoxyprotein intermediate has been formed. Preferred epoxy compounds for use in conjunction with olefinically unsaturated monomers are of the general formula wherein R 1 is a mono- or poly unsaturated alkenyl radical, an alkenyloxy radical or an unsaturated acyl radical, preferably one containing a CO group conjugated with a carboncarbon double bond, R 2 is a hydrogen atom or a lower alkyl radical (up to C 4 ) and n is 0, 1 or 2. Many compounds are particularly mentioned. Preferred monomers are the water-soluble derivatives of vinyl alcohol, acrylic acid and methacrylic acid including the amides, nitriles and esters of acrylic and methacrylic acid. The carrier may be crosslinked either during its formation or after its reaction with the epoxy compound-protein product. The examples are directed to the preparation of carrier-bound proteins from the following constituents: glucose oxidase, uricase or hexokinase; acrylic acid - (2,3 - epoxy - propyl ester), 1 - (allyloxy)- 2,3 - epoxypropane or 1,2 - epoxybut - 3 - ene; acrylamide (carrier substance); N,N1 - methylene - bis - acrylamide (crosslinking agent); and 3 - dimethyl - amino - propionitrile and ammonium peroxydisulphate (polymerization initiator combination). The polymerizates are forced through a metal sieve of 0À5 mm. mesh size and washed to remove adsorptively bound enzyme.