Patent GB-1441543-A (GB)
Title: Immobilized enzyme compositions
IPC: C12N11/08
Filed: 1974-04-24
Abstract: 1441543 Insolubilized enzymes DENKI KAGAKU KOGYO KK 24 April 1974 [30 May 1973 (2)] 17924/74 Heading C3H [Also in Division C2] A water-insoluble enzyme composition comprises an enzyme or cells of micro-organisms containing enzymes bound to a hydrophilic, water-insoluble anion-exchange resin, said resin being a polymer which contains a quaternized pyridine ring in the polymer unit and preferably having an anion-exchange capacity of 2-5 meq./g. The pyridene unit is derived from 2- vinylpyridine (2-VP), 4-vinylpyridene (4-VP), 2 - methyl - 5 - vinylpyridene, 5 - methyl - 2- vinylpyridene, 3 - allylpyridene or 1 - vinylquinoline which is copolymerized with one or more of monomers selected from styrene (St), alpha - methylstyrene, halogenated styrenes, ethylene, propylene, acryl acid, methacrylic acid, acrolein, methylmethacrylate (MMA), vinylacetate, acrylonitrile, vinylchloride, acrylamide, butadiene, isoprene, chloroprene, divinylbenzene (DVBz), ethylene glycol dimethacrylate (EDMA), butane diolacrylate, polyethylene glycol dimethacrylate, diallylphthalate and methylenebis - (acrylamide) and, optionally, with polymers selected from polybutadiene, polyisoprene, polychloroprene, styrene/butadiene copolymer, polypropylene, chlorinated polyethylene and polyvinylalcohol. The copolymers are prepared by random, block or graft. polymerization, depending on the physical form required for the final product. When random polymerization is used, cross-linking monomers must be present to prevent the copolymer dissolving on quaternization (VP, 20-99 mole per cent; divinyl compounds, 0À5-30 mole per cent) whereas this is not necessary for graft and block copolymers (VP, 25-75 mole per cent). The base polymer is quaternized with methyl chloride, methyl bromide, methyl iodide and propyl bromide preferably, but other agents may be used (including sulphur di- and trioxides, thionyl chloride, dimethyl sulphate; acid chlorides and anhydrides; metal halide compounds and acidic compounds having high molecular weights). Enzymes specified for insolubilization are protease, aminoacylase, adenosine deaminase, AMP deaminase, amidase, α-amylase, #-amylase, glucoamylase, succinyloglucoamylase, lactic dehydrogenase, trypsin, papain, ribonuclease, dextranase, glucose oxidase, penicillin acylase, chymotrypsin, ficin, pepsin, carboxypectidase, streptokinase, urease, invertase, maltase, lactase, lipase, cellulase, catalase, melibiase, tyrosinase, aspartase, glucose isomerase, phenol oxidase, racemase and cells of microorganisms which retain the activities of said enzymes. Quaternized base polymers prepared are: P1-P3, 2-VP/St, P4-P5, 2-VP/St/DVBz, P6, 4-VP/MMA/DVBz, P7, 2-VP/EDMA/St, P8, 4-VP/St/DVBz, P9, 2-VP/MMA/DVBz, (P1-P3 are block copolymers and P4-P9 random copolymers). Insolubilized enzyme compositions exemplified are: Ex. 1-5, aminoacylase+P1, P2, P4- P6, Ex. 6-11, glucoamylase+P1, P2, P5-P8, Ex. 12-15, glucose isomerase+P4, P6, P8, P9, Ex. 16, glucose oxidase+P5, Ex. 17-18, alkaline protease+P4, P6, Ex. 19, invertase+ P5, Ex. 20-23, succinyl glucoamylase+P5- P8. The compositions have activities analogous to the free enzyme and are used in the conversions of various substrates (see Division C2).