PREPARATION OF ACTIVATED SUPPORTS CONTAINING LOW PK AMINO-GROUPS - A NEW TOOL FOR PROTEIN IMMOBILIZATION VIA THE CARBOXYL COUPLING METHOD

A method for the preparation of new aminated agarose gels containing m onoaminoethyl-N-aminoethyl structures, MANA-agarose gels, has been dev eloped. These gels contain primary amino groups with a very low pK val ue (6.8). In addition to that, we have been able to prepare very highl y activated gels (e.g., 10% agarose gels containing up to 200 muEq of primary amines per milliliter). These two properties make these activa ted supports suitable for performing novel and interesting methods for protein immobilizations via very mild carbodiimide activation of carb oxy groups. For example, very effective coupling reactions can be perf ormed at pH 5.0-6.0 in the presence of low concentrations of activatin g agent, e.g., 1 mm. By using a model industrial enzyme, beta-galactos idase from Aspergillus oryzae, we have been able to demonstrate the ex cellent prospects of these novel activated supports.